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Bacopa monnieri

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Propagation
Cultivation
Morphology
Phytochemistry
Improvement
History & Society
Work Log

[1] [2] [3]

Propagation

Germination

media germination temperature °C note reference
         

[4]

Vegetative

[5] [6]

In-Vitro

basal media supplements source target note reference
           

[7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21] [22] [23] [24] [25] [26] [27] [28] [29] [30] [31] [32] [33] [34] [35] [36] [37] [38] [39] [40] [41] [42] [43] [44] [45] [46] [47] [48] [49] [50] [51] [52] [53] [54] [55] [56] [57] [58] [59] [60] [61] [62] [63] [64] [65] [66] [67] [68] [69] [70] [71] [72] [73] [74] [75] [76] [77] [78] [79] [80] [81] [82] [83] [84] [85] [86] [87] [88] [89] [90] [91] [92] [93] [94] [95] [96] [97] [98] [99] [100] [101] [102] [103] [104] [105] [106] [107] [108] [109] [110] [111]zollingeriana [112] [113] [114] [115] [116] [117] [118] [119] [6]

Cultivation

Planting density (m-2) inter-row space (cm) intra-row space (cm) note reference
         

[120] [121] [122]

Harvest

[123] [124] [125]

Yield

product source yield per season (kg/ha) note reference
         
product source yield per plant note reference
         

[126] [127] [15] [128] [129] [4] [130] [131] [132] [133] [134] [135] [136] [137] [138] [139]

Soilless

NFT and DFT hydroponics strongly promote monnieri production with DFT outperforming on most growth measures. Moderate effects were noted on the concentration of secondary metabolites, though this might be due to the dilution effect of increased growth. Regardless, a 6-10 fold increase in harvest dry weight was seen over soil-based controls.[129]

[129] [140] [141] [142]

Soil

soil type pH C-content % precipitation temperature (°C) altitude (m) note reference
               

[126] [127] [120] [143] [144] [130] [131] [132] [133] [134] [145] [136] [146] [137] [138] [147]

Fertilization

type rate time note reference
         

[126] [148] [127] [128] [149] [150] [143] [144] [131] [151] [152] [136] [138] [139] [153] [142] [154] [155]

Temperature

Lighting

fixture type photoperiod illumination note reference
         

[127] [15] [26]

Pests

[156] [157] [149] [145] [139] [142] [155] [154] [147]

Ecology

[148] [158] [159] [128] [156] [157] [160] [150] [143] [144] [161] [151] [152] [135] [119] [145] [146] [138] [139] [162] [122] [153] [142] [154] [155] [147]

Morphology

character measurement unit notes reference
         

[11] [12] [41] [43] [55] [163] [67] [164] [165] [166] [167] [168] [135] [169] [122] [6]

Roots

[165]

Stem

[166] [168]

Leaves

[166] [168]

Inflorescence

Seeds

Phytochemistry

compound source concentration (mg/g dry weight) note reference
         

[7] [148] [11] [13] [15] [18] [170] [57] [68] [171] [96] [172] [125] [139]

Infraspecific Variation

[173] [174] [5] [164] [175] [108] [176] [167] [133] [177] [134] [169] [137]

Biosynthesis

[8] [158] [21] [26] [27] [28] [29] [30] [31] [32] [33] [34] [35] [37] [42] [157] [45] [46] [48] [49] [178] [160] [58] [63] [64] [66] [67] [70] [72] [73] [75] [150] [76] [78] [83] [86] [87] [89] [92] [143] [94] [101] [144] [103] [110] [114] [117] [121] [132] [118] [135] [122] [147] [6]

Distribution

[178] [55] [166] [168] [165] [169]

Timecourse

[130] [133]

Improvement

trait improvement status reference
     

Identification

variety description reference
     

[173] [179] [180] [181] [174] [5] [175] [182] [183] [107] [184] [185] [176] [167] [133] [177] [134] [186] [169] [137]

Inheritance & Stability

[8] [13] [51] [68] [81] [82] [84] [92] [95]

Methods

type note reference
     

[9] [187] [49] [51] [188] [56] [59] [70] [88] [89] [97] [106] [107] [113] [125]

History & Society

[189] [149] [190]

Work Log

03 Sep 2023

Started cuttings in DWC hydroponics.

12 Jul 2023

06 Mar 2022

Started an unknown number of seeds. They are so tiny, that it is hard to tell the difference between seeds and chaff. I used the remainder of the 30-pack of seeds from 2020 (SM10585).

Sowed on filter paper in a petri dish, wrapped in plastic wrap, and placed in the incubator at 30°C.

Bibliography

  1. Nandy, Samapika and Mukherjee, Anuradha and Pandey, Devendra Kumar and Dey, Abhijit, Bacopa Monnieri: The Neuroprotective Elixir from the East—Phytochemistry, Pharmacology, and Biotechnological Improvement, pp. 97--126, 2020. doi: 10.1007/978-981-15-1394-7_2.
    Bacopa monnieri (L.) Wettst. (Scrophulariaceae), commonly known as water hyssop, is an extensively used herb in Ayurveda. The bioactive phyto-constituents like the dammarane-type triterpenoid saponins such as bacosides A, B, and C were reported mostly for neuroprotective, nootropic, and memory-enhancing properties. Furthermore, B. monnieri also acts as a natural antioxidant, which can ameliorate morphine dependence, memory deficit in epilepsy, hepatocarcinogenesis, hepatotoxicity, β-amyloid cytotoxicity, inflammation, and oxidative stress (Singh et al., Physiol Plant. 2019). The neuroprotective role of the plant was manifested against neurodegenerative disorders like Parkinson’s disease (PD), Alzheimer’s disease (AD)-associated dementia, attention-deficit hyperactivity disorder in children, and anxiety. Somer report also found the traces of heavy metals in various species of Bacopa monnieri. Inadequate knowledge regarding underlying mechanism of neuroprotection and associated systematic structure-activity relationship (SAR) studies and development of semisynthetic derivatives of saponins are of high importance to facilitate rational designing of novel drugs based on leads from bacopasaponins. This review systematically summarizes the origin, analysis of the pharmacotherapeutic properties, and structural significance of bioactivities exerted by the bacopasaponins. Moreover, future research arenas, lacunae in the present understanding, are also highlighted in this work. Together, this book chapter may further direct to the synthetic optimization of bacopasaponins as lead compounds for the designing of efficacious and clinically acceptable analogues.
  2. Mallick, B., A Short Review on Physico-Chemical Properties of Bacopa Monnieri L, International Journal of Medicinal Plants. Photon, vol. 110, pp. 735--741, January 2016.
    Material of our interest in the present study is the medicinally important Brahmi (Bacopa monnieri L.) herb. The herb has been so chosen because of its various pharmaceutical and medicinal properties. Since the important elemental constituents, phases and complexes of the medicinal plant possess different curative capability of human disease. So, it is important to know the details of the above herb, its chemicals composition, pharmaceutical and medicinal components, important elements and material phases present are defined in this mini-review.
  3. Suman, Shikha, Assessment of Bacopa Monnieri Plant: An Economically Important Herb, International Journal of Industrial Biotechnology and Biomaterials, vol. 1, no. 1, 2015. doi: 10.37628/jibb.v1i1.26.
    Assessment of Bacopa monnieri Plant: An Economically Important Herb
  4. Thapliyal, Chetna, Effect of Growth Hormones on Germination Seedling Propagation and Biomass Production of Medicinal Plants Bacopa Monnieri L Penell and Withania Somnifera Dunal, University, 2009. url: http://shodhganga.inflibnet.ac.in:8080/jspui/handle/10603/184102.
    newline
  5. Koul, A. and Kushwaha, M. and Gupta, A. and Mallubhotla, S., Regenerative Potential and Phytochemical Diversity Among Five Accessions of Bacopa Monnieri (l.) Wettst, undefined, 2015. url: https://www.semanticscholar.org/paper/REGENERATIVE-POTENTIAL-AND-PHYTOCHEMICAL-DIVERSITY-Koul-Kushwaha/a7dbae43fe3551cd132b66213a2db9090bb08cf6.
    Biochemical profiling proved the qualitative differences in the carbohydrate, alkaloid, saponin and terpenoid contents in leaf, shoot and root samples of all accessions, and LC-ESI-MS analysis revealed that the shoot extract of accession BM002 contains highest Bacoside content as compared to that of its root extract. Distant geographical niches have shown pronounced effect on variability in organoleptic characteristics as well as biochemical constituents between accessions of a given plant species. The present study has been designed to evaluate the regenerative potential of five accessions of Bacopa monnieri, a herb which is of high commercial potential with a legendary reputation as a memory vitalizer through morphogenetic, phytochemical and Liquid chromatography- Electrospray ionization-Mass spectra (LC-ESI-MS) analysis. Culture media supplemented with exogenous plant hormones had a varied effect on in vitro culture morphogenesis and biomass production in the accessions. Maximum friable callus was regenerated in accession BM003 (GI: 54.61 ± 0.11) on B5 media supplemented with 2,4-D (1.0 mg l -1 ) while MS media with NAA (1.0 mg l -1 ) + BAP (0.5 mg l -1 ) resulted in maximum indirect organogenesis in BM003 (GI: 99.58 ± 0.06). Biochemical profiling proved the qualitative differences in the carbohydrate, alkaloid, saponin and terpenoid contents in leaf, shoot and root samples of all accessions. LC-ESI-MS analysis revealed that the shoot extract of accession BM002 (7236.38 µg gm -1 ) contains highest Bacoside content as compared to that of its root extract (1015.32 µg gm -1 ) and the shoots
  6. Sagar, R. and Suman, S. and Kapur, P., Changes in Functional Traits of Bacopa Monnieri Due to Water Treatment, 2012. url: https://www.lap-publishing.com/catalog/details/store/gb/book/978-3-659-22954-1/changes-in-functional-traits-of-bacopa-monnieri-due-to-water-treatment.
    Results showed inhibition of flowering in water limitation, this inhibition was less in micropropagated plant than the vegetative plant, and fruiting was never seen in this plant during its life cycle. Study of plant functional traits is essential to predict the nature of plant response to different biotic and abiotic perturbations. In this book, the effects of water treatment on the morphological, physiological and biochemical traits of micro as well as vegetative propagated Bacopa monnieri are reported. Results showed inhibition of flowering in water limitation, this inhibition was less in micropropagated plant than the vegetative plant. Fruiting was never seen in this plant during its life cycle. Further, the pigments, plant biomass, NPP, RGR, NAR and bacoside contents were maximum at the intermediate age of the micropropagated plant in daily watering condition, which could be suitable time for the harvesting of the plant for maximum active compounds. Amongst the traits; SLA, LAR, NAR, chlorophyll, NPP, P and reducing sugar synergstically explained the variation in bacoside content. When the studied four sets of B. monnieri were allowed to grow with herbaceous species in field condition, study showed difference in species composition could be due to variation in traits of B. monnieri. This invited different weed management practices of B. monneri in different treatments.
  7. Sahoo, Krishna Kalyani and Sahu, Binod B. and Singh, V. R. and Patra, Nivedita, Enhanced Production of Bacopa Saponins by Repeated Batch Strategy in Bioreactor, Bioprocess and Biosystems Engineering, February 2022. doi: 10.1007/s00449-022-02700-4.
    Cultivation of cell suspension culture of Bacopa monnieri targeting the production of bacosides was explored in a 5-l stirred tank reactor using statistically optimized conditions. The bioreactor cultivation conditions were modified and this led to profuse biomass growth (2.81\,±\,0.20~g/l) and total bacosides (1.26\,±\,0.23~mg/g in cells and 0.60\,±\,0.11~mg/l in fermenter broth) production in 9~days. The values of static volumetric mass transfer coefficient (kLa), dimensionless mixing time (Nm) were measured in the bioreactor. The culture grew efficiently and produced enhanced amount of bacoside A (5.59\,±\,0.41~mg/g total bacosides in cells and 3.12\,±\,0.13~mg/l in the fermenter broth) using one cycle of repeated batch strategy adopted in the bioreactor for 15~days. The intracellular concentration of bacoside A3 (1.18\,±\,0.11~mg/g), bacopaside II (2.09\,±\,0.35~mg/g), bacopaside X (0.79\,±\,0.17~mg/g) and bacopasaponin C (2.24\,±\,0.23~mg/g) were significantly higher in repeated batch as compared to batch bioreactor cultivation. The yield of total bacosides in the fermenter broth was 5-times higher in repeated batch as compared to batch cultivation. This strategy can be helpful for the enhanced production of other valuable triterpenoid saponins.
  8. Pramanik, Biswajit and Sarkar, Sutanu and Bhattacharyya, Somnath and Gantait, Saikat, Meta-Topolin-Induced Enhanced Biomass Production via Direct and Indirect Regeneration, Synthetic Seed Production, and Genetic Fidelity Assessment of Bacopa Monnieri (L.) Pennell, a Memory-Booster Plant, Acta Physiologiae Plantarum, vol. 43, no. 7, pp. 107, June 2021. doi: 10.1007/s11738-021-03279-1.
    The present study reports enhanced in vitro mass propagation, synthetic seed production and its regeneration, acclimatization, and genetic fidelity assessment of in vitro regenerants of Bacopa monnieri (L.) Pennell; a medicinal plant renowned for its memory-enhancing property. For the first time, meta-Topolin (mT) as well as N6-benzyladenine (BA) and kinetin at their variable concentrations (0.5, 1. 1.5, 2 and 2.5~mg/l) were supplemented individually in Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) basal medium to induce multiple shoots from shoot tip explants, initially collected from two-month-old vegetatively propagated plants. The best result in terms of multiplication (\textasciitilde\,7 shoots/explant) and biomass accumulation (\textasciitilde\,451.3~mg fresh weight,\,\textasciitilde\,104.3~mg dry weight) was obtained in MS medium supplemented with 1~mg/l mT. For rooting of in vitro shoots, supplementation of 1.5~mg/l IAA and 0.5~mg/l IBA in MS media resulted in induction of the maximum number (\textasciitilde\,20) and highest length (\textasciitilde\,5.5~cm) of roots, respectively. Most efficient indirect regeneration and biomass production from green and hard basal calli (induced spontaneously in BA-supplemented media) was recorded in a simple MS medium. Synthetic seeds were produced from in vitro shoot tips using 2.5\% sodium alginate and 75~mM calcium chloride solutions and were successfully regenerated in half-strength MS medium. Six-week-old in vitro regenerants were transferred to different substrates for acclimatization, among which cocopeat was found to be the best exhibiting the highest survival and growth in external conditions. Finally, molecular marker-(ISSR) and phenology-based genetic fidelity assessment of the in vitro regenerants exhibited that the in vitro regenerants were true-to-type in nature within themselves as well as with the mother plant.
  9. Sujipuli, K. and Inthima, P. and Yimtragool, N. and Warnnissorn, Netnaphis and Warnnissorn, P. and Prasarnpun, S., In Vitro Colchicine-Induced Polyploids from Different Explant Segments of Bacopa Monnieri, 2021. doi: 10.2306/SCIENCEASIA1513-1874.2021.003.
    Findings indicated that colchicine-induced tetraploids could have beneficial uses for genetic improvements to increase the medicinal value of local Bmo herbal medicine powder production in Thailand. Bacopa monnieri (Bmo) is a highly valued medicinal plant, widely used for several pharmacological products in Thailand. The diploid progenitor of Bmo grown in the wild as a natural source has low biomass and bacoside contents. Fifteen treatment combinations between three Bmo explant types (apical shoot, node and leaf segments) and five various colchicine concentrations (0.00, 0.05, 0.075, 0.1, and 0.5\% w/v) were determined for a number of polyploid inductions, including bacoside contents and gene expression. Results showed that low concentration (0.05\%) of colchicine was more effective in inducing multiple shoots (17.88 regenerants per explant) and tetraploid plantlets (15\%) from leaf segments than other treatment combinations. Tetraploid clone (no.4x-3) showed the highest BmoOSC gene expression, bacoside-A3 (4.276±0.019 mg/g dry weight) and bacoside-C contents (5.040±0.078 mg/g dry weight) compared to diploid progenitors and mixoploid plants. These findings indicated that colchicine-induced tetraploids could have beneficial uses for genetic improvements to increase the medicinal value of local Bmo herbal medicine powder production in Thailand.
  10. Koul, A. and Sharma, Munish and Sharma, Shagun and Sharma, Jyotsna and Verma, V. and Mallubhotla, S., Exploration of Therapeutic Potential of Bacopa Monnieri Using in Silico Approach and Optimization of Culture Conditions for Bacoside Production, Journal of Proteomics \& Bioinformatics, vol. 14, no. 5, pp. 536, 2021. url: https://www.semanticscholar.org/paper/Exploration-of-Therapeutic-Potential-of-Bacopa-in-Koul-Sharma/45576517e3743a32aa9a0a39cb69cd6fa12e07e0.
    The docking of the bacoside A3 molecule is attempted to find the binding mechanism with the suitable target which will be helpful in understanding the molecular mechanism involved in the development of Alzheimer’s disease. Bacopa monnieri plant extract has been used as a nerve tonic in ancient Indian folklore since ages and bacosides are the main key component behind neuro-potential of the herb. Accumulated β-amyloid protein causes the hyperphosphorylation of tau protein which in turn is the cause of tangled nerve projections in Alzheimer’s brain. However, the interaction of bacosides with amyloid protein has not been studied so far. Computational approach including docking analysis showed hydrogen bonding of bacoside molecule with important amino acid residues present in amyloid protein which proved the potential of bacoside A 3 in targeting Alzheimer’s. In addition to this, in vitro culture conditions have demonstrated their critical influence on secondary plant metabolism. Kinetic studies conducted in our experiment showed that cell suspension cultures of bacopa placed under shaking light conditions produce more bacoside content as compared to other conditions. Besides these properties, bacopa plants have also been known to improve the chemical characteristics of saline and clayey soils thus, enhancing the growth of agriculturally important crops. In this work, we attempted the docking of the bacoside A3 molecule to find the binding mechanism with the suitable target which will be helpful in understanding the molecular mechanism involved in the development of Alzheimer’s disease. Up-scaling of memory enhancer bacoside molecule has also been attempted under in vitro conditions that should deliver maximum benefits with minimal tissue loss during harvestation.
  11. Tauk, Chanraksmey and Chuaduangpui, Pornpimon and Khawniam, Tassanee, Effects of Explants on Plant Regeneration and Concentration of Paclobutrazol on Morphological Responses of Dwarf Water Hyssop (Bacopa Monnieri), Naresuan University Journal: Science and Technology (NUJST), vol. 29, no. 3, pp. 56--66, February 2021. doi: 10.14456/nujst.2021.26.
    Water hyssop is an ornamental aquatic plant which is very popular among aquarium hobbyists. The present study reports on the effect of PBZ concentration on water hyssop to produce dwarf aquatic plants. The different types of explant were cultured on MS medium supplemented with 0.5 mg/L BA for shoot and root induction. The single nodal was cut and cultured on MS supplemented with various PBZ concentrations to produce the dwarf ornamental aquatic plant. The results showed that shoot induction started within 2 weeks and the maximum average number of shoots received from the node with leaf explant was 12.11 shoots after 4 weeks of culturing. The highest concentration at 40 mg/L of PBZ retarded stems, number of leaves, number of roots, root length, and the survival rate declined within 2 weeks of culturing. Only 2.5 mg/ L PBZ stimulated shoot induction at 51. 85\%, but 20 mg/ L of PBZ the percentage of shoot induction of 48.15 was obtained. Shoot length did not increase after treating with 40 mg/ L of PBZ and also resulted in the lowest average number of leaves of 1.25 leaves. The maximum concentration of PBZ caused toxic reactions, such as yellowing, stunted leaves, short roots, and death of in vitro cultures. The shoot length decreased with increasing concentrations of PBZ. Thus, at 2.5 mg/ L of PBZ was suitable for producing dwarf water hyssop plant with the maximum shoot length, maximum number of leave and longest root. Finally, the dwarf plantlets were acclimatized successfully to 4 weeks in aquarium conditions.
  12. Sarkar, Sayantika and Jha, Sumita, Effects Associated with Insertion of Rol Genes on Morphogenic Potential in Explants Derived from Transgenic Bacopa Monnieri (L.) Wettst, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 146, no. 3, pp. 541--552, September 2021. doi: 10.1007/s11240-021-02092-5.
    The present study deals with the establishment of rolA-transgenic and rolB-transgenic plants for the first time through Agrobacterium tumefaciens mediated transformation, exploiting the inherent morphogenic potential of an important medicinal plant, Bacopa monnieri (L.) Wettst. The rolA-transgenic and rolB-transgenic plants showed integration and expression of rolA and rolB genes respectively, whereas Ri-transformed plants showed integration and expression of rolA, rolB, rolC and rolD genes. Morphogenic potential of different types of explants derived from rolA-transgenic, rolB-transgenic and Ri-transformed plants on basal medium was evaluated. Shoot organogenesis was enhanced significantly in leaf (1.6-fold) and internode (1.4-fold) explants derived from rolA-transgenic plants, rolB-transgenic leaf (2.4-fold) and internode (1.6-fold) explants as well as leaf (5.2-fold) and internode (3.3-fold) explants derived from Ri-transformed plants compared to explants from non-transformed plants. Substantial increase in root organogenesis was also noticed in rolA-transgenic leaf (1.7-fold) explants, rolB-transgenic leaf (3.6-fold) and internode (1.4-fold) explants as well as leaf (4.1-fold) and internode (1.9-fold) explants derived from Ri-transformed plants compared to non-transformed ones. In addition to this, growth of root tip and shoot regeneration was also noticed from Ri-transformed root explants, but not in rolA-transgenic, rolB-transgenic and non-transformed roots. Clones of each transgenic plant line harboring rol genes depicted notable phenotypic changes including reduced shoot and internode length, increased number of nodes/plant, leaves/plant and roots/plant. The leaf morphology was altered in rolB-transgenic and Ri-transformed plants but not in rolA-transgenic plants.
  13. Tamboli, Firoj A. and Rangari, Vinod D. and More, Harinath N. and Kutwade, Vijay H. and Patil, Vivek M., Antidepressant Effects of Natural and Micropropagated Bacopa Monnieri (L.) Plant Extracts, Indian Journal of Pharmacy and Pharmacology, vol. 8, no. 2, pp. 141--145, June 2021. doi: 10.18231/j.ijpp.2021.022.
    Background: Bacopa monnieri Linn. an important medicinal plant in Indian system of alternative medicine belonging to Scrophulariaceae family. It is distributed in the wet and marshy lands throughout India, Nepal and many other parts of world. This study was carried out to evaluate the natural and micropropagated Bacopa monnieri plant extract for antidepressant activity. Materials and Methods: Antidepressant effects of natural and micropropagated plant extract was evaluated by spontaneous motor activity. All the extracts were administered 30 min prior to the test. The standard drug chlorpromazine (2mg/kg) was used. The locomotor activity was performed using an actophotometer after different doses of test drug. Result: The locomotor activity of control and treated mice were recorded and statistically correlated among the control, standard and the test drugs. The inhibition of spontaneous motor activity in case of BMN, BMS extract was found 291, 93.5 and 277, 93.66 count at interval of 30 and 60 min. respectively. But in case of BMM plant extract it was found 290, 87.33 and 146 count at 30, 60 and 120 min. interval respectively so results showed that a significant antidepressant-like effect at a dose of 100 mg/kg. It can be used as adjuvant therapy for depression. Conclusion: There was statistically highly significant (p value Keywords:~Bacopa monnieri, Natural plant, Micropropagated plant, Spontaneous Motor activity,~Actophotometer, Anti-depressant effect.
  14. Rahe, Md Adnan and Mollika, Sanjida Rahman and Khan, M. S. and Tanjina and Banu, A. Afroos and Amin, G. and Habib, M. and Akter, S. and Islam, Mousona and Sharmin, R., PTC \& B In Vitro Micropropagation of Bacopa Monnieri ( L . ) Penn .-An Important Medicinal Plant, 2020. url: https://www.semanticscholar.org/paper/PTC-%26-B-In-vitro-Micropropagation-of-Bacopa-(-L-.-)-Rahe-Mollika/2469d665ff86961ede650cf5fb44ca6aa686aa5b.
    Investigation on in vitro multiple shoot regeneration in Bacopa monnieri (L.) Penn. using leaf and nodal explants was carried out on MS containing various concentrations and combinations of BAP, Kn, NAA and 2,4-D. Of the two explants, leaf showed the best response towards shoot regeneration and subsequent plant development on MS with 1.0 mg/l BAP and 0.25 mg/l Kn. In this combination, the mean number of shoots/explant was 10.6 ± 0.11 in leaf and 9.6 ± 0.29 in nodal explants. Maximum shoot length was recorded as 12.6 ± 0.21 and 11.20 ± 0.30 from leaf and nodal explants after six weeks of culture, respectively. Half strength of MS supplemented with 0.25 mg/l IBA was found to be the best medium for root formation. The in vitro regenerated plantlets were successfully transplanted in soil after acclimatization.
  15. Aggarwal, Akanksha and Mathur, Ashwani, Nexus between Light and Culture Media on Morphogenesis in Bacopa Monnieri and Saponin Yield Thereof, Heliyon, vol. 6, no. 10, pp. e05245, October 2020. doi: 10.1016/j.heliyon.2020.e05245.
    Bacopa monnieri, a well-documented nootropic plant of high commercial global demand had been explored for its effect in alleviating other diseases and symptoms. This is primarily attributed to different phytocompounds present in the plant. One of the major constituents among them are saponins. However, variation in agro-climatic conditions and choice of germplasm often affect the growth rate and yield of phytocompounds that significantly impact the efficacy of the plant and its extract. Tissue culture has been attempted to improve the yield of phytocompounds but is often restricted by higher cost and scalability. Current study explores the role of commercial hydroponic media ‘Leafy 200’ vis-a -vis Murashige and Skoog (MS) media, under different color and intensity of lights, on plant morphogenesis, biomass and saponin yield. Blue light induced more shoot differentiation than normal white light. Statistical studies performed using fractional factorial design showed no significant variations in the yield of saponins among the extracts. The study suggests that hydroponic culture to be a sustainable solution and possible substitute to tissue culture that may be exploited for scalable cultivation of the plant.
  16. Chauhan, R. and Shirkot, P., Micropropagation of Endangered Medicinal Plant Bacopa Monnieri (L.) Pennell, undefined, 2020. url: https://www.semanticscholar.org/paper/Micropropagation-of-endangered-medicinal-plant-(L.)-Chauhan-Shirkot/110dc71159a9f8e8527afbbc4f5aad0c4d2b791a.
    A protocol for micropropagation of selected genotype of Bacopa monnieri (L.) Pennell has been standardized and it is found that this herb helps in improving intelligence, memory and learning ability. Bacopa monnieri (L.) Pennell, commonly known as “Brahmi” is an important medicinal herb of the family Scrophulariaceae. It has main importance as tonic for nervous disorders and mental diseases. It helps in improving intelligence, memory and learning ability. In the present work, a protocol for micropropagation of selected genotype of Bacopa monnieri (L.) Pennell has been standardized. Axillary buds were used as explants for in vitro plant regeneration studies in Bacopa monnieri (L.) Pennell. Axillary shoot buds showed best in vitro establishment (87.50\%) on MS medium supplemented with 0.5 mg/l BAP and 0.5 mg/l Kn. Best in vitro shoot multiplication was obtained on MS medium supplemented with 1.0 mg/l BAP and 0.5 mg/l Kn. For in vitro rooting, MS medium supplemented with 0.4 mg/l IAA was found to be the best medium (93.33\%). Regenerated plantlets were successfully acclimatized by using cocopeat followed by a mixture of soil: sand: FYM in ratio 1:1:1.
  17. Sharma, N. and R, G. and Pandey, Ruchira and Agrawal, A., Influence of Explant Types, Non-Embryogenic Synseed and Reduced Oxygen Environment on in Vitro Conservation of Bacopa Monnieri (L.) Wettst, In Vitro Cellular \& Developmental Biology - Plant, 2020. doi: 10.1007/s11627-020-10078-9.
    This is the first report on success of combining two slow-growth strategies of reduced oxygen environment and synseed technology for midterm (12 mo) in vitro conservation of germplasm. The effect of using encapsulated shoot tips (ST) and nodal segment (NS) explants, in combination with mineral oil (MO) overlay, for in vitro regeneration and efficient midterm conservation of Bacopa monnieri (IC468878), a medicinal herb with potent nervine properties is described. On a standard multiplication medium [Murashige and Skoog (MS) containing 0.2 mg l−1 benzyladenine (BA)] after 8 wk of culture, the number of shoots regenerated from ST (17.50\,±\,0.50) to NS (19.10\,±\,1.20) were not significantly different; however, the shoot length regenerated from NS (7.30\,±\,0.70 cm) was significantly higher than ST (2.50\,±\,0.30 cm). Encapsulation of ST explants did not have any significant effect on the number of shoots (18.10\,±\,1.00) and shoot length (3.50\,±\,0.30 cm), while there was significant decrease in shoot number (10.30\,±\,1.0) and shoot length (1.50\,±\,0.40 cm) in NS explants. In general, use of MO overlay resulted in highly significant reduction of growth and increase in survival of cultures. The combined strategy of encapsulation and MO overlay led to growth reduction in terms of number of shoots (2.50\,±\,0.30 in ST, 2.30\,±\,0.20 in NS) as well as shoot length (0.50\,±\,0.02 cm in both explants). After 12-mo in vitro conservation, encapsulated NS explants in combination with MO overlay gave the highest survival rate (91.6\,±\,5.7\%) and number of shoots (21.4\,±\,1.6). This is the first report on success of combining two slow-growth strategies of reduced oxygen environment and synseed technology for midterm (12 mo) in vitro conservation of germplasm.
  18. Saha, P. and Sarkar, S. and Jeyasri, Rajendran and Muthuramalingam, Pandiyan and Ramesh, M. and Jha, S., In Vitro Propagation, Phytochemical and Neuropharmacological Profiles of Bacopa Monnieri (L.) Wettst.: A Review, Plants, 2020. doi: 10.3390/plants9040411.
    Bioreactor based micropropagation was found to increase the multiplication rate of shoot cultures for the commercial propagation of B. monnieri plants and the maximum content of bacosides has been recorded in shoot biomass using an airlift bioreactor system. Bacopa monnieri has been used as a reputed drug in the Indian traditional ayurvedic system for centuries. This medicinal herb with important phytopharmaceuticals has been popularly known as “Brahmi”. In recent years, B. monnieri has been extensively studied for its bioactive constituents, constituents responsible for memory enhancing effect, and also its diverse other useful effects. It possesses many pharmacological activities such as antioxidant, gastrointestinal, endocrine, antimicrobial, anti-inflammatory etc. The plant has been also used for the treatment of neurological and neuropsychiatric diseases. Due to its multipurpose therapeutic potential, micropropagation using axillary meristems and de novo organogenesis has been extensively studied in the species and is being reviewed. High frequency direct shoot organogenesis can be induced in excised leaf and internode explants in the absence of exogenous phytohormones and the rate of induction is enhanced in the presence of exogenous cytokinins, supplements, growth regulators, etc. Using explants from tissue culture raised plants, direct shoot regeneration leading to production of more than 100 rooted plants/explant within 8–12 weeks period with 85\%–100\% survival in the field after acclimatization can be expected following optimized protocols. Bioreactor based micropropagation was found to increase the multiplication rate of shoot cultures for the commercial propagation of B. monnieri plants. The maximum content of bacosides has been recorded in shoot biomass using an airlift bioreactor system. Further studies for the biosynthesis of bacosides and other secondary metabolites need to be conducted in the species utilizing untransformed shoot cultures in bioreactors.
  19. Raj, Poornima and Anbumalarmathi, J. and Sharmili, S. Aruna, In Vitro Propagation of Bacopa Monnieri (L.), Biotechnology Journal International, pp. 32--39, July 2020. doi: 10.9734/bji/2020/v24i430111.
  20. Rahe, Md Adnan and Mollika, Sanjida Rahman and Khan, M. Salim and Banu, Tanjina Akhtar and Amin, GM Al and Habib, Md Ahsan and Akter, Shahina and Islam, Mousona and Sharmin, Ripa Akter, In Vitro Micropropagation of Bacopa Monnieri (L.) Penn. - An Important Medicinal Plant, Plant Tissue Culture and Biotechnology, vol. 30, no. 1, pp. 57--63, June 2020. doi: 10.3329/ptcb.v30i1.47791.
    Investigation on in vitro multiple shoot regeneration in Bacopa monnieri (L.) Penn. using leaf and nodal explants was carried out on MS containing various concentrations and combinations of BAP, Kn, NAA and 2,4-D. Of the two explants, leaf showed the best response towards shoot regeneration and subsequent plant development on MS with 1.0 mg/l BAP and 0.25 mg/l Kn. In this combination, the mean number of shoots/explant was 10.6 ± 0.11 in leaf and 9.6 ± 0.29 in nodal explants. Maximum shoot length was recorded as 12.6 ± 0.21 and 11.20 ± 0.30 from leaf and nodal explants after six weeks of culture, respectively. Half strength of MS supplemented with 0.25 mg/l IBA was found to be the best medium for root formation. The in vitro regenerated plantlets were successfully transplanted in soil after acclimatization. Plant Tissue Cult. \& Biotech. 30(1): 57-63, 2020 (June)
  21. Koul, Anuja and Mallubhotla, Sharada, Elicitation and Enhancement of Bacoside Production Using Suspension Cultures of Bacopa Monnieri (L.) Wettst, 3 Biotech, vol. 10, no. 6, pp. 256, May 2020. doi: 10.1007/s13205-020-02242-0.
    The effect of different elicitors [jasmonic acid, salicylic acid] and precursors [calcium pantothenate, cholesterol, sodium nitroprusside] on the stimulation of biomass and secondary metabolite production in suspension cultures of Bacopa monnieri was studied. Induction of primary callus cultures was successfully carried out on the Gamborg’s B5 (B5) medium fortified with 2, 4-D (1.0~mg~l−1) using Bacopa monnieri leaves as explants. The friable fine suspension cell culture was raised on parent B5 media without agar. The elicitation using different elicitors and precursors at varying concentrations was carried out over a period of 3, 6, 9, 15~days. Elicitor treated cultures showed marked increase in biomass and bacoside production around 6th–9th day (0.98 GI DW). In the present study, salicylic acid at 1.0~mg~l−1 induced a maximum elicitation in bacoside content (6.58~mg~g−1 DW). The present study provides favorable evidence on the potential of bacoside production using suspension cultures of B. monnieri. The study results also indicate the beneficial effects of elicitation on metabolite production in in vitro suspension cultures of B. monnieri plant known for its cognitive improving properties.
  22. Sape, Subba Tata and Kandukuri, Aditya Vardhan and Owk, Aniel Kumar, Direct Axillary Shoot Regeneration with Nodal Explants of Bacopa Monnieri (l.) Pennell – a Multi Medicinal Herb, Journal of Applied Biological Sciences, vol. 14, no. 2, pp. 190--197, August 2020. url: https://www.jabsonline.org/index.php/jabs/article/view/726.
    The aim of the present investigation is to develop an optimized culture medium for direct induction of adventitious multiple shoots from nodal explants of Bacopa monnieri an important medicinal herb. The nodal explants were excised from the in vivo B. monnieri and were inoculated over different media fortified with cytokinins BAP or KIN alone or in combination with IAA for in vitro response. The rooting was established with Murashige and Skoog media containing with auxin NAA. All the treatments are performed in a controlled aseptic chamber and maintained the cultures at temperature 22ºC for 4 and 2 weeks respectively for shoot induction and rhizogenesis. The results are expressed in mean of three replicates with standard error. Adventitious multiple shoots proliferated from nodal explants cultured on all the tested media. The highest percent shoot induction, shoot number per node and shoot length was observed on MS medium containing BAP(1.5mg/l) and IAA(0.5mg/l). The excised shootlets cultured on MS media fortified with various levels of NAA and identified 0.2mg/l NAA was best for maximum rhizogenesis. The rooted plants were successfully transferred to the field. Overall, the present study showed superior results for shoot number per node over previous reports. Therefore, it is hoped that this report will contribute to our knowledge about in vitro mass propagation of medicinal herb B. monnieri.
  23. Tata, Sape Subba, Development of a Profused In Vitro Shoot Multiplication Using Leaf Explants of Bacopa Monnieri (L.) Pennell, Current Botany, pp. 14--17, April 2020. doi: 10.25081/cb.2020.v11.6030.
    Bacopa monnieri (L.) Pennell is an important medicinal plant used for the preparation of medhyarasayan (rasayana). Leaf explants of field grown young plants of B. monnieri was used to establish an efficient regeneration protocol with cytokinin (BAP) and auxin (IAA). The highest multiplication, i.e. (220 shoots/leaf, a cumulative of 2200 shoots from 10 explants) were noticed after 45 days of culture in MS medium supplemented with BAP(1.5mg/L) and IAA(0.5mg/L). The optimum concentration of growth regulator for shoot elongation and rooting was recorded in MS+GA3 (0.25mg/L) and MS+IBA (1.5mg/L) respectively. The rooted plantlets were successfully established in green house conditions.
  24. Kumari, Ritika, Induction of Callus from Different Explants of Bacopa Monnieri and Effect of Adjuvant on the Growth Rate of the Calli., Indian Journal of Scientific Research, vol. 10, no. 1, pp. 113--121, August 2019. url: https://go.gale.com/ps/i.do?p=AONE&sw=w&issn=09762876&v=2.1&it=r&id=GALE%7CA632332152&sid=googleScholar&linkaccess=abs.
  25. Samanta, Dipu and Mallick, Bidisha and Roy, Debleena, In Vitro Clonal Propagation, Organogenesis and Somatic Embryogenesis in Bacopa Monnieri (L.) Wettst, Plant Science Today, vol. 6, no. 4, pp. 442--449, October 2019. doi: 10.14719/pst.2019.6.4.600.
    Bacopa monnieri (L.) Wettst is a well-known medicinal herb in the Ayurveda. It is also used as laxative and curative for ulcers, inflammation, anaemia, scabies, leucoderma, asthma and epilepsy, enlargement of spleen, leprosy and others. In vitro propagation and regeneration through somatic embryogenesis of B. monnieri has played an important role in the production of healthy, disease-free plants with desirable traits. In B. monnieri, there are few reports which indicate rapid regeneration and somatic embryogenesis. For in vitro clonal propagation, the highest shoot formation was obtained when BAP 2 mg/ l used. The best response for rooting was obtained in IAA 1.0 mg/ l. The recorded survival rate of the plants was 70\%. Plants were without any detectable phenotypic variations. Cytological study indicated that the chromosome number remain same (2n= 64) in in vitro and in vivo roots. A rapid, simple and efficient protocol for plantlet regeneration was achieved through embryogenic callus from leaf explants of B. monnieri. Callus induction and embryogenesis were significantly affected by presence/absence and type and concentration of growth regulators. Best organogenic callus induction was obtained in MS medium supplemented with BAP 5mg/ l. For induction of somatic embryogenesis, auxin (2, 4-D 1 mg/ l) was used in the culture medium subsequently in basal media for embryo maturation. Kn 0.2 mg/ l was the best for production of plantlet from embryo. Thus, this can be an easiest protocol for stable clonal propagation and plant regeneration through somatic embryogenesis in B. monnieri. The protocol used here for propagation and regeneration is much easier, low cost and reliable.
  26. Watcharatanon, Kawissara and Ingkaninan, Kornkanok and Putalun, Waraporn, Improved Triterpenoid Saponin Glycosides Accumulation in in Vitro Culture of Bacopa Monnieri (L.) Wettst with Precursor Feeding and LED Light Exposure, Industrial Crops and Products, vol. 134, pp. 303--308, August 2019. doi: 10.1016/j.indcrop.2019.04.011.
    Bacopa monnieri (L.) Wettst is widely used in Ayurveda treatment for improving memory and reducing anxiety. Triterpenoid saponin glycosides are the bioactive compounds in this plant. This study aimed to investigate the influence of precursor feeding and LED light exposure on the enhancement of bioactive compounds in in vitro cultures of B. monnieri. Precursor feeding l-alanine and l-phenylalanine significantly enhanced the accumulation of triterpenoid saponin glycosides in all treatments. The optimal duration for precursor feeding was obtained after treatment for 6 days. Total triterpenoid saponin glycosides production increased 2.4-fold to 46.98\,±\,0.19\,mg/g dry wt. and 2.6-fold to 49.41\,±\,0.12\,mg/g dry wt. by adding 5\,mM l-alanine and 150 μM l-phenylalanine for 6 days, respectively. In addition, triterpenoid saponin glycosides reached the maximum level (58.53\,±\,0.74\,mg/g dry wt.) after treatment with a combination of precursors (1.0\,mM l-alanine and 100 μM l-phenylalanine for 6 days). Furthermore, blue and red light on day 28 of treatment increased triterpenoid saponin glycosides (24.11\,±\,0.07, 22.18\,±\,0.19\,mg/g dry wt.) 1.7-fold and 1.5-fold higher than that obtained with white light. This study indicated that bioactive compounds in B. monnieri in vitro cultures can be enhanced by feeding precursors and LED light exposure. These techniques can be applied at the industrial level of crops and food supplements.
  27. Lala, Sanchaita, Enhancement of Secondary Metabolites in Bacopa Monnieri (L.) Pennell Plants Treated with Copper‐based Nanoparticles in Vivo, IET Nanobiotechnology, vol. 14, no. 1, pp. 78--85, December 2019. doi: 10.1049/iet-nbt.2019.0124.
    The study aims to document the effect of starch‐stabilised copper‐based nanoparticles (CuNPs) on the biosynthesis of pharmaceutically valuable secondary metabolites, especially saponins, of the reputed nootropic herb Bacopa monnieri (L.) Pennell. CuNPs were synthesised chemically by the reduction of cupric sulphate pentahydrate with ascorbic acid using starch as the capping agent. They were characterised by UV–visible spectrophotometry, Fourier‐transform infra‐red spectroscopy, X‐ray diffraction, high‐resolution transmission electron microscopy and zeta potential. The nanoparticles consisted of cuprous oxide and metallic copper, were approximately spherical, polydispersed with diameter {$<$}20~nm. Hydroponically grown B. monnieri plants were treated in vivo with the CuNPs between the concentrations of 0–100~mg l−1. Spectrophotometric estimation of the total contents of saponins, alkaloids, phenolics, flavonoids and DPPH radical scavenging capacity from the methanolic extracts of the whole plants showed a hormetic increase in the content of secondary metabolites in a concentration‐dependent manner from 5~mg l−1 until it declined at toxic metabolic concentration. This was accompanied by an increase in ROS markers hydrogen peroxide and malondialdehyde as well as a hormetic effect on activities of phenylalanine ammonia lyase and antioxidant enzymes catalase, ascorbate peroxidase and superoxide dismutase. CuNPs at sub‐toxic concentrations were found to enhance secondary metabolism and antioxidant capacity in Bacopa monnieri through ROS‐mediated defence response.
  28. Murthy, M. and Khandayataray, Pratima and Samal, D. and Laha, R., Effects of Surface Sterilizing Agents, Sucrose and Plant Growth Regulatory Hormone Concentration Levels on Micropropagation of Bacopa Monnieri L., undefined, 2019. url: https://www.semanticscholar.org/paper/Effects-of-Surface-Sterilizing-Agents%2C-Sucrose-and-Murthy-Khandayataray/72b8b1c53ba3b6d99d8577b8ac6ebf963b34e332.
    Higher number of roots and increased root length was observed in MS Media + Sucrose (30 g/L) + Agar (7 g/l), and it was found that in the medium MS, rapid callus growth was observed which turned pale yellow and of globular appearance, and the combination of 2,4-D, KIN and BAP were found good for rapidcallus growth, shoot length and multiple shoots respectively. Background and Aims: Bacopa monnieri L. is used to treat sleep deprivation and anxiety. Microropagation is rapid and helps in exsitu preservation of this endangered plant. Methods: Three surface sterilizing agents with different concentrations [ethanol (EtoH; 50 and 70\%), mercuric chloride (HgCl2; 0.1 and 0.5\%) and sodium hypochloride (NaOCl; 0.1, 0.5 and 1\%)] were used at different time intervals (1, 3, 5, 8 and 10 minutes) with and without hot water to obtain the good aseptic culture. Various concentrations of sucrose (30, 10 and 20 g/L) plus agar 7 g/L was used to observe the effect on root initiation and length in Bacopa monnieri L. The initiated explants Original Research Article Murthy et al.; AJRIB, 2(2): 1-12, 2019; Article no.AJRIB.48761 2 (leaf, node and internode) were cultured on MS media and supplemented with different combination of 2, 4-D (2, 4-dichlorophenoxyacetic acid), KIN (Kinetin) and BAP (6-Benzyl amino purine), IAA (Indole-3-acetic acid), NAA (1-Naphthaleneacetic acid) and GA3 (Gibberellin A3) to induce callus, to initiate root and root length, to initiate auxillary bud induction and length and to multiple shoots after inoculation. Results: It has been seen that 0.1\% of HgCl2 (Mercuric chloride) followed by warm water showed 100\% aseptic culture conditions. MS media with sucrose (30 g/L) along with agar (7 g/L) showed multiple root initiation whereas MS media with 2,4-D (0 mg/L), KIN (2.0 mg/L) and BAP (2.0 mg/L); NAA (1.0 mg/L) + (KIN 1.0 mg/L) + BAP (1.0 mg/L) and BAP (2.0 mg/L) + KIN (1.0 mg/L) + GA3 (0.5 mg/L) were found good for rapid callus growth, shoot length and multiple shoots respectively. Conclusions: In conclusion, higher number of roots and increased root length was observed in MS Media + Sucrose (30 g/L) + Agar (7 g/L). It was also found that in the medium MS + 2,4-D (0 mg/L), KIN (2.0 mg/L) and BAP (2.0 mg/L) rapid callus growth was observed which turned pale yellow and of globular appearance. Highest shoot length was observed in MS + NAA (1.0 mg/L) + (KIN 1.0 mg/L) + BAP (1.0 mg/L) medium. Maximum number of multiple shoots was found in MS + BAP (2.0 mg/L) + KIN (1.0 mg/L) + GA3 (0.5 mg/L).
  29. Dogan, Muhammet, Effect of Cadmium, Chromium, and Lead on Micropropagation and Physio-Biochemical Parameters of Bacopa Monnieri (L.) Wettst. Cultured in Vitro, Rendiconti Lincei. Scienze Fisiche e Naturali, vol. 30, no. 2, pp. 351--366, June 2019. doi: 10.1007/s12210-019-00796-7.
    Heavy metals are released into the environment as a result of anthropogenic activities such as melting metallic ores, industrial productions, power stations, and pesticides. Among these, cadmium (Cd), chromium (Cr) and lead (Pb) are the most common heavy metals in the environment and can cause serious toxic effects on both animals and plants. In this study, the effect of Cd, Cr, and Pb on the propagation of Bacopa monnieri (L.) Wettst. by tissue culture was investigated. In addition, various physio-biochemical parameters (photosynthetic pigments contents, protein contents, lipid peroxidation, and hydrogen peroxide) and survival frequency of the plant were evaluated. The shoot tip explants of B. monnieri were cultured in Murashige and Skoog (MS) nutrient media supplemented with 0.50~mg/L 6-benzylaminopurine (BAP) and different concentrations of Cd, Cr, and Pb (0.25–1.00~mg/kg) for 10~weeks. 100\% shoot regeneration was determined in MS medium containing 0.25, 0.50, and 0.75~mg/kg Cd, 0.25~mg/kg Cr and 0.25 and 0.50~mg/kg Pb. Mean number of shoots per explant and shoot lengths decreased significantly with increasing metal concentration. In Cd, Cr, and Pb applications, maximum number of shoots per explant was obtained as 30.52, 22.47, and 27.12 shoot/explant and the highest shoot lengths were determined as 4.16, 4.27, and 4.35~cm, respectively. As the concentration of the metal exposed to the explant increased, the photosynthetic pigment contents of the plant decreased. The negative impact of metals on photosynthetic pigment contents was found to increase in the order: Cd\,{$<$}\,Cr\,{$<$}\,Pb. Similarly, the protein content of plants also decreased due to the negative effect of metals. On the other hand, malondialdehyde (MDA) and hydrogen peroxide (H2O2) values elevated with increasing concentrations of heavy metals. The findings of this study carried out under in vitro conditions may shed light on researchers who want to investigate the effects of heavy metals on plants.
  30. Ahmed, Aziz and Ahmad, Sayeed and Shuaib, Mohd and Ahmad, Rizwan and Somani, Neelam and Sharma, Ankita, Effect of Auxin and Cytokinin on Regeneration, Total Phenolics and in Vitro Antioxidant Activities of Bacopa Monnieri (L.) Pennell, Journal of Ayurvedic and Herbal Medicine, vol. 5, no. 2, pp. 54--59, June 2019. doi: 10.31254/jahm.2019.5205.
    Context: Cells are protected from cellular harm by antioxidant compounds. These compounds secure cells from h Cells are protected from cellular harm by antioxidant compounds. These compounds secure cells from harmful free radicals which initiated oxidative stretch. Bacopa monnieri (L.) Pennell (Schropulariaceae) have medicinal properties, such as sedative, tranquilizing, antioxidant and memory-enhancing effects. Objective: To determine the total phenolic content and concentrations of flavonoids and also in vitro antioxidant activity in methanol extract of shoot culture from leaves explants of Bacopa monnieri (L.) Pennell supplemented with various combinations of auxin and cytokinin hormone. Material and method: Murashige and Skoog (MS) media was selected for the experiment and different concentration of auxin and cytokinin were added in media. Leaves explants were inoculated on media for callus initiation. Indirect shoot formation occurred on calli of leaf explants after subculture. Total phenolic, flavonoid content and in vitro antioxidant activities such as DPPH radical scavenging activity, hydrogen peroxide (H2O2) scavenging activity and ferric reducing power of the shoot cultures are quantitatively estimated by UV spectroscopy. Result: The highest content of total phenolics (3.14g/100g d.w.) and flavonoids (0.565 g/100g d.w.) were obtained in shoots regenerated on the MS medium supplemented with BA (2mg/lit) and IAA (1mg/lit). Shoot culture on MS medium supplemented with BA (2mg/lit) and IAA (1mg/lit) showed the highest DPPH radical scavenging activity (85.31\%), highest H2O2 scavenging activity (75.31\%) and highest ferric reducing power (25.3µg/g). Discussion and conclusion: Excellent in vitro antioxidant activity was given by extract of Shoot culture on MS medium supplemented with BA (2mg/lit) and IAA (1mg/lit). So they can be used as natural antioxidants after isolation and purification.
  31. Bhardwaj, Pragya and Jain, Chakresh Kumar and Mathur, A., Comparative Analysis of Saponins, Flavonoids, Phenolics and Antioxidant Activities of Field Acclimatized and in Vitro Propagated Bacopa Monnieri (L.) Pennell from Different Locations in India, undefined, 2019. url: https://www.semanticscholar.org/paper/Comparative-analysis-of-saponins%2C-flavonoids%2C-and-Bhardwaj-Jain/51d4f0d6328d6af7c780b12968c41a00a653da26.
    The studies may help understanding the role of environmental and in vitro propagation conditions in regulating biosynthesis of therapeutically important phytocompounds better, and thereby useful in developing a scalable process. Bacopa monnieri (L.) Pennell, a well-documented nootropic plant, commonly called ‘herb of grace’ and traditionally Brahmi, is extensively used in herbal formulations for neurological disorder. However, variations in active phytopharmaceutically important compounds in the formulations often affect their therapeutic efficacy and market acceptance. In this study, we compared pharmaceutically important phytocompounds viz. saponins, flavonoids and phenolics along with antioxidant activities in field acclimatized B. monnieri plants from different geographical locations in India. Results have shown comparatively higher saponins, phenolics and flavonoids yield in BM-7 (field acclimatized from Haridwar, Uttranchal) and higher antioxidant activities in BM-4 (field acclimatized from Ghaziabad, Uttar Pradesh). The soil samples of the plants sources have also shown variations in the macronutrient compositions. In comparison, when propagated in in vitro conditions using four different culture media, all plants respond differently with comparatively higher dry weights in Murashige and Skoog medium (1962). Further, analyses of the phytocompounds in MS medium revealed variations in the phytocompounds yield and antioxidant activities. While BMT-3 (Jammu) and BMT-6 (Lucknow) reported around 9 to 11 fold increase in saponins yield compared to field acclimatized plants, BMT-2 (Delhi) showed 10 and 12 fold increase in total phenolic content and antioxidant activities, respectively. The studies may help understanding the role of environmental and in vitro propagation conditions in regulating biosynthesis of therapeutically important phytocompounds better, and thereby useful in developing a scalable process.
  32. Aasim, Muhammad and Sameeullah, Muhammad and Karataş, Mehmet and Bakirci, Seyma and Bakhsh, Allah and Akhtar, Mohd Sayeed, An Insight into Biotechnological Approaches Used for the Improvement of Secondary Metabolites from the Medicinal Aquatic Plant, Water Hyssop (Bacopa Monnieri L.), pp. 123--152, 2019. doi: 10.1007/978-981-13-7438-8_5.
    Water has a significant role on human civilizations by providing food and water along with plants grown on the banks or within the water bodies. Aquatic plants or semiaquatic plants are the group of plants that love to live in or near water bodies, and some of these plants are economically important due to their usage as food or medicinal plants. Water hyssop or brahmi (Bacopa monnieri L.) is one of the important semiaquatic/aquatic plants that has been used for medicinal purposes since ancient time in Indian subcontinent. The plant contains several secondary metabolites like bacosides which are used as memory enhancer tonic commercially. Brahmi-based registered drugs are available in India and other countries as memory enhancer tonic and for other diseases like Alzheimer’s disease, anxiety, asthma, stomach ulcers, and respiratory ailments and for curing chronic diseases like cancer. Bacopa is facing the threat of extinction from wild as it is not a cultivated plant and propagation through seed is limited due to low availability of and viability of seeds. On the other hand, extensive works on the propagation of this important medicinal plant has been reported to develop in vitro protocols for its conservation and plant propagation for secondary metabolite production. Different in vitro techniques like cell suspension culture, callus culture, and organogenesis have been reported with the objective of producing or enhancing bacoside. Furthermore, application of other biotechnological approaches like Agrobacterium-mediated genetic transformation studies, use of mutagens, and in vitro polyploidization have also been reported. Thus, the aim of this chapter is to highlight the application of different biotechnological approaches used for the production, conservation, and secondary metabolite production of B. monnieri.
  33. Bhardwaj, Pragya and Goswami, Navendu and Narula, Pankhuri and Jain, Chakresh Kumar and Mathur, Ashwani, Zinc Oxide Nanoparticles (ZnO NP) Mediated Regulation of Bacosides Biosynthesis and Transcriptional Correlation of HMG-CoA Reductase Gene in Suspension Culture of Bacopa Monnieri, Plant Physiology and Biochemistry, vol. 130, pp. 148--156, September 2018. doi: 10.1016/j.plaphy.2018.07.001.
    Bacopa monnieri (L.) Wettst. is a well documented nootropic plant, extensive known for alleviating symptoms of neurological disorder, along with other symptomatic relief. This property is attributed to the active phytocompounds, saponins (bacoside A) present in the plant. However, lack of stringent validation guidelines in most of the countries bring to the market, formulations differing in phytocompounds yield, thereby suggesting possible variation in therapeutic efficacy. The in-vitro suspension cultures of the Bacopa monnieri, provide an ease of scale-up, but regulating saponin yield is a stringent task. The aim of the study is to explore the effects of different concentrations (0, 0.25, 0.50, 0.75 and 1.0\,ppm) of zinc oxide nanoparticles (ZnO NP) (24\,nm in size), in regulating growth rate, bacoside yield and transcriptional profile of HMG CoA reductase gene in the suspension cells of Bacopa monnieri. Results showed a linear correlation between Bacoside A yield and ZnO NP concentrations with around 2 fold increase in total bacoside A concentration at 1\,ppm. Also, ZnO NP supplemented suspension cells showed variation in the specific growth rate. Neuroprotective properties, analyzed using methanolic extracts of suspension cells again obtrude the extract of ZnO NP supplemented (0.75\,ppm and 1\,ppm) culture for better response in alleviating oxidative stress mediated damage to neuronal cells. ZnO NP supplemented system showed lower expression of HMG CoA reductase gene (the rate limiting step in bacoside A biosynthesis) but higher concentration of bacoside A, suggesting possible role of ZnO NP in isoprenoid pathway than MVA pathways.
  34. Leonard, J. and Seth, Bishwanath and Sahu, Binod B. and Singh, V. R. and Patra, Nivedita, Statistical Optimization for Enhanced Bacoside A Production in Plant Cell Cultures of Bacopa Monnieri, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 133, no. 2, pp. 203--214, May 2018. doi: 10.1007/s11240-017-1373-6.
    The bioactive compound, bacoside A, has immense importance for the treatment of memory disorders and Alzheimer’s disease. Due to the growing commercial interest in the herb, Bacopa monnieri, it has been listed as highly endangered species. The present study was aimed at enhancing the production of bacoside A using an alternative technology of plant cell suspension culture. Initial experiments of docking simulations using bacoside A showed good inhibition of acetyl cholinesterase (binding energy value of −\,20~kcal/mol), when comparison was made with other phytocompounds and the synthetic drug for Alzheimer’s disease. In vitro experiments established that B. monnieri cell suspension culture can be developed in Murashige and Skoog medium containing containing 0.1~mg/L benzylaminopurine and 0.5~mg/L naphthalene acetic acid. Plackett–Burman studies predicted that the most effective factors for maximum biomass production were inoculum size (t-value of 4.87), sucrose concentration (t-value of 0.25) and KH2PO4 concentration (t-value of 0.007). The nitrate to ammonium ratio (t-value of −~0.42) did not have significant effect on the cell suspension biomass. The optimum concentration of the crucial variables obtained from a central composite design were—inoculum size of 2~g/L, sucrose concentration of 30~g/L and KH2PO4 concentration of 1.24~mM in one-sixth strength MS medium. The best model for optimum production of biomass and bacoside A was experimentally verified and the correlation between the predicted and actual values was found to be 99\% for biomass and 94\% for bacoside A production. The experimental results have been discussed in the present work.
  35. Sahoo, K. K., Optimization of Bacoside a Production in Cell Suspension Culture of Bacopa Monnieri for Scale-Up Using Stirred Tank Reactor, undefined, 2018. url: https://www.semanticscholar.org/paper/Optimization-of-Bacoside-a-Production-in-Cell-of-Sahoo/38fe6c296b709030f635f217b08eb770677109ce.
    An economical alternative method is outlined for the propagation and conservation of Bacopa monnieri that provides commercially feasible method for the large scale production of bacoside A. Bacopa monnieri(Brahmi), an amphibious plant growing in tropical climate, has been reported to be a source of the secondary metabolite,bacoside A,known for the regeneration of nerve cells in the brain.As a result of its excessive demand in the pharmaceutical industry, cultivation of this plant has gained wide popularity.In order to augment the supply of this drug from natural origin,alternative plant cell culture techniques need to be developed.Field-grown plantlets of high-yielding variety of Bacopa monnieri were utilized in this study to develop high-yielding callus cultures for the mass production of bacoside A.The explant type was optimized for utmost callus biomass production. Substrate inhibition kinetics was studied to determine the inhibitory concentrations of major limiting substrates. Using response surface methodology,the phytohormone concentrations were optimized for the improved synthesis of biomass and bacoside A.The optimized factors were utilized to study the batch and repeated-batch cultivation of Bacopa monnieri in stirred tank bioreactor.A combination of bavistin treatment (0.5\% solution for 5 minutes)and sodium hypochlorite treatment (1\% solution for 5 minutes)was found to effectively sterilize the field-grown explants for invitro culture development.Rooting of the shoots was induced in MS medium containing 1 mg/l IAA in 7 days. Callus was induced from leaf explants in MS medium containing 0.5 mg/l NAA and 0.1 mg/l BAP within 10 days. Leaf explants were found to be the most ideal explant for callusinduction. 1.0 mg/l NAA and 0.1 mg/l BAP were the optimum concentration for the induction of callus from leaf explant; and 0.2 mg/l NAA and 1.2 mg/l BAP were the optimized concentrations for the maintenance of the callus. The growth of the cells in suspension culture was observed to get inhibited at 120 g/l sucrose (carbon source) and 1.25 g/l KH2PO4 (phosphate source). Using the optimized conditions,batch cultivation of Bacopa monnieri was carried out in a stirred tank reactor. 18.97 g/l DW biomass and 18.05 mg/g DW bacoside A was produced by using cell suspension as inoculum for batch cultivation in stirred tank reactor in 6 days, whereas, 3.04 g/l DW biomass and 27.36 mg/g DW bacoside A was produced by using pre-established callus culture as inoculum in the reactor in 9 days. Repeated-batch cultivation of Bacopa monnieri in stirred tank reactor yielded 1.40 g/l DW biomass in 13 days. Hence this study outlines an economical alternative method for the propagation and conservation of Bacopa monnieri. Also, it provides commercially feasible method for the large scale production of bacoside A.
  36. Kashyap, Suman and Kale, Radha D., Micropropagation of B. Monnieri Using Body Fluid of Earthworms (Coelomic Fluid) as Plant Tissue Media, Vegetos- An International Journal of Plant Research, vol. 31, no. 1, pp. 104, 2018. doi: 10.5958/2229-4473.2018.00015.0.
  37. Goel, Aanchal and Kaur, Amanpreet and Kumar, Anil, Biochemical and Histological Changes during in Vitro Rooting of Microcuttings of Bacopa Monnieri (L.) Wettst., Acta Physiologiae Plantarum, vol. 40, no. 3, pp. 64, March 2018. doi: 10.1007/s11738-018-2641-8.
    The present study was conducted to investigate the biochemical changes vis-à-vis histological changes during adventitious rooting of microcuttings of Bacopa monnieri (L.) Wettst. The rooting in these microcuttings was induced on basal MS medium and medium supplemented with different concentrations of indole-3-acetic acid and indole-3-butyric acid (IBA). Presence of lower auxin concentration (1.0~µM) in the medium enhanced rooting and significantly improved number of roots per shoot but maximum root length was observed on basal MS medium. Histological studies were conducted to identify different phases of rooting in these microcuttings. The root meristemoids with distinct polarity become visible after 3~days and mark the beginning of in vitro root initiation phase. It was followed by primordia elongation, root emergence and visible rooting on the 5th day of culture on medium supplemented with auxins. Biochemical studies were also conducted from basal portions of microcuttings cultured on MS medium supplemented with 1.0~µM IBA and control (basal MS medium) from 0 to 7~days. Total carbohydrate content was lower during initial periods (up to day 1) and was found to increase during root initiation and primordia development, which reflects high energy demands for active cell divisions. A significantly higher level of phenols was recorded in microcuttings on medium supplemented with IBA. Polyphenol oxidase, peroxidase (POX), ascorbate peroxidase activities were also found to vary during different phases of rhizogenesis. Early phases were also marked with the lower activities of POX and IAAO. This study revealed significant role of enzymes, sugars and phenols during different phases of rooting.
  38. Ranjan, R. and Kumar, Shailesh and Singh, A., An Efficient in Vitro Propagation Protocol of Local Germplasm of Bacopa Monnieri (L.) Found in Bihar: A Plant with Wide Variety of Medicinal Properties, undefined, 2018. url: https://www.semanticscholar.org/paper/An-efficient-in-vitro-propagation-protocol-of-local-Ranjan-Kumar/0da53a69fb13f81cf6346417ee82c30ba85b3318.
    An effort has been made to develop a rapid and reproducible protocol for the in vitro mass multiplication through callus induction of Bacopa monnieri through calledus induction from leaf and nodal segment for regeneration. Bacopa monnieri L. Penn. is an important medicinal crop. The problems with natural propagation is death of seedlings at two leaved stage, short viability of seed, marshy areas requirements and slow growth of stem cutting. Therefore, in the present investigation, an effort has been made to develop a rapid and reproducible protocol for the in vitro mass multiplication through callus induction of Bacopa monnieri. Nodal and leaf segments were used as explants for regeneration. A friable, compact, globular, morphogenic, green and white colored callus was initiated in different combination of media. For callus induction from leaf segments, MS basal media supplemented with 0.5 mg/l BAP and 0.5 mg/l NAA, supported the best result (75\% callus induction) and MS salt supplemented with 4.0 mg/L BAP and 0.5 mg/L IAA supported best callus responses for nodal segment (85\% callus formation). The MS salt supplemented with 1.0 mg/L BAP+ 0.5 mg/L NAA gave best results for shooting and multiple shooting induction from callus of leaf and nodal segment. Shoot induction percentage (85\%), average number of shoots (15) and average shoot length (9.5 cm) was higher in case of callus developed from nodal segment as compared to leaf segments. Best performing rooting media (MS + 20g/l sucrose and 7g/l agar) induced rooting in 80\% of shoots. Two weeks after hardening plants were transferred to soil in field/open conditions and 98\% survival rate was observed, and the regenerated plantlets showed robust growth.
  39. B, Rajendra Prasad and M, Venkateshwarlu and G, Odelu and Rao.m, Madan Mohan G. {and} Babu, An Efficient IAA and BAP-Assisted in Vitro Micropropagation through Shoot Tip Culture of Bacopa Monnieri (L.) Wettst. – A Medicinally Important Plant, International Journal of Pharma and Bio Sciences, vol. Volume 9 Issue 4, 2018 (October - December). doi: 10.22376/ijpbs.2018.9.4.b30-37.
    Archives issue of International Journal of Pharma and Bio Sciences which aims to cover the latest outstanding developments in the field of pharmaceutical and biological sciences
  40. Ranjan, Rajeev and Kumar, Shailesh, A Rapid In Vitro Propagation Protocol of Local Germplasm of Bacopa Monnieri (L.) Induced through Direct Organogenesis from Nodal Explants, Journal of Pharmacognosy and Phytochemistry, vol. 7, no. 1, pp. 2515--2518, 2018. url: https://www.phytojournal.com/archives/2018.v7.i1.2963/a-rapid-in-vitro-propagation-protocol-of-local-germplasm-of-bacopa-monnieri-l-induced-through-direct-organogenesis-from-nodal-explants.
    The natural propagation of local germplasm of Bacopa is affected by poor seed viability, death of seedlings at two leaved stage, marshy areas requirements and slow growth of stem cutting. An efficient in vitro propagation protocol through direct organogenesis from nodal explants has been reported here for local germplasm of Bacopa monnieri. For axillary bud break and shoot proliferation from nodal segments, MS basal media supplemented with 1.0 mg/l BAP and 1.0 mg/l IAA, supported the best response (85\% induction) and the same media combination also offered the best result for multiple shoot formation. The average number of shoots (10) and average shoot length (8.2 cm) was observed from nodal segment. Best performing rooting media (MS + 20g/l sucrose and 7g/l agar) induced rooting in 81\% of shoots. The average number of roots (6) and average root length (3.1 cm) was observed from in vitro regenerated shoot. Two weeks after hardening plants were transferred to soil in field conditions and 90\% survival rate was observed, and the growth of hardened plantlets was vigorous.
  41. Almusawi, Abdulminam Hussien Ali and {Al-Aradi}, Haleemah Jabbar and Hammadi, Kadhim Jasim, The Impact of Salinity Stress on Morphological and Anatomical Aspect of Water Hyssop Bacopa Monnieri (L.) Wettst Grown in Vitro, African Journal of Biotechnology, vol. 16, no. 15, pp. 801--807, April 2017. doi: 10.5897/AJB2016.15699.
    This study was conducted at Marine Science Center, Basrah University, Basrah, Iraq during the period of 2015 to 2016. In the present work, we used NaCl and polyethylene glycol (PEG) to induced abiotic stress in vitro. Results show that high shoot numbers (24) were produced on MS (solid) medium enriched with 0.8 g/L NaCl in addition to cytokinins while, inclusion of drought agent PEG to proliferation medium (solid and liquid) produced a minimum shoots of 5 and 11 respectively. Root growth and development were also investigated in this study. It was found that high root development was observed on MS (solid or liquid) medium free of hormone (control) and MS (solid or liquid) medium containing 0.8 g/L NaCl 26.67, 25, 21.67 and 18 respectively whereas, less root development was obtained on MS medium (solid or liquid) enriched with NaCl and 26 g/L PEG. In this study it was found that inclusion of the amino acid proline to the stressed media did not improve the growth of shoots or roots. Histological analysis of multiplied shoots under abiotic stress agents showed accumulation of insoluble starch granules in the parenchyma cells of the cortex layer and this may be a part of protective tolerate mechanism used by medicinal hyssop (Bacopa monnieri) against abiotic stress. Key words: Bacopa monnieri, medicinal hyssop, Brbin, micropropagation, salt stress, polyethylene glycol (PEG), proline.
  42. Manoj Godbole, Poornananda Madhava Naik, The Effect of Heavy Metals on In Vitro Adventitious Shoot Production and Bacoside A Content in Bacopa Monnieri (L), Mapana - Journal of Sciences, vol. 14, no. 4, pp. 1--10, July 2017. doi: 10.12723/mjs.35.1.
    In the present study the effect of heavy metals (MnSO 4, ZnSO 4 and CuSO 4 ) on adventitious shoot regeneration, biomass and bacoside A accumulation in Bacopa monnieri was carried out. The leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg l -1 kinetin (Kin) with varying concentration of heavy metals (Control: Mn-0.10 mM, Zn-0.03 mM and Cu- 0.0001mM; Mn: 0.20, 0.40, 0.80 and 1.60 mM; Zn: 0.06, 0.12, 0.24 and 0.48 mM; Cu: 0.02, 0.05, 0.10 and 0.20 mM). Optimum number of adventitious shoots (123.50 shoot/explants), fresh weight (3.826 g) and dry weight (0.226 g) of Bacopa monnieri were obtained in the medium with 0.12 mM Zn concentration. The highest production
  43. Sarkar, Sayantika and Jha, Sumita, Morpho-Histological Characterization and Direct Shoot Organogenesis in Two Types of Explants from Bacopa Monnieri on Unsupplemented Basal Medium, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 130, no. 2, pp. 435--441, August 2017. doi: 10.1007/s11240-017-1231-6.
    In the present study, high frequency regeneration has been obtained via de novo direct shoot organogenesis from leaf and internode explants in Murashige and Skoog (MS) basal medium without any phytohormone supplementation in Bacopa monnieri, an indigenous traditionally used medicinal herb. Leaves and internodes from different positions were excised from 4-weeks-old in vitro propagated B. monnieri plants and cultured on MS basal medium supplemented with 3\% (w/v) sucrose and 0.75\% (w/v) agar for 4 weeks. The induction of de novo shoot buds was observed at petiolar cut edges of leaf and both proximal and distal cut ends of internode explants within 10–15 days of culture. The first histological changes could be observed after 4–5 days, with meristematic activity of vascular bundles. Proliferation of epidermal cells gave rise to dome-shaped protuberances followed by shoot apical meristems formation and their vascular connections with explant tissues within 2 weeks of culture. However, a basipetal gradient of shoot regeneration from both types of explants collected along the branch axis was noticed after 4 weeks of culture. Leaf and internode explants near the basal region exhibited significantly higher number of shoot buds and micro shoots (8.8/leaf explant and 15/internode explant). Microshoots (7–12 micro shoots/leaf or internode explants) elongated (shoot length 8–9~cm) within 8 weeks on phytohormone free MS medium. Excised micro shoots rooted (100\%) in hormone free MS medium within two weeks of culture. Rooted plants were then acclimatized and transferred to field with 95\% survival. This protocol may be used for micropropagation, genetic transformation as well as a model system for evaluation of changes associated with acquisition of competence of differentiated cells in phytohormone free medium.
  44. Kashyap*, Suman and Kapoor, Neera and Kale, Radha D., Micropropagation of B. Monnieri Using Humin Media in Plant Tissue Culture, Annals of Plant Sciences, vol. 6, no. 05, pp. 1625--1629, May 2017. doi: 10.21746/aps.2017.05.004.
    Humus is a complex material formed during the breakdown of organic matter. Earthworm castings contain a high percentage of humus. These humic substances are capable of improving plant nutrition and growth, reminiscent of hormones. Residue obtained after acid-base treatment of vermicompost was used as plant tissue culture media for the micropropagation of Bacopa monnieri. Tukey’s Studentized Range (HSD) Test has clearly indicated that development of leaves from nodes was significantly higher in humin alone without any supplements. The F value was 5.5 and Pr {$>$} F was 0.0087. The weight of the plantlets (in milligrams) was significantly higher in the humin medium and humin supplemented with vitamins and micronutrients but was least in the humin medium with growth regulators. The difference observed was at the level of {$<$} 0. 0001. The explant development on humin alone has shown the significant percentage survival of in vitro micropropagated plantlets when compared with other treatments. Explants responded maximum to humin only but not so on adding the supplements and the level of significance was at Pr {$>$} F was {$<$} 0.0001. The maximum growth stimulatory effect was found in aqueous extract of vermicompost which can be made out from the Student’s t-test performed to compare the means of weight of plantlets grown on humin and vermicompost extract media with and without chemical supplements respectively. The probability of this result, assuming the null hypothesis is 0.0014.
  45. Mohammed, N., Mathematical Model for Enhanced Production of Bacoside Using Batch Cultivation for Bacopa Monnieri, undefined, 2017. url: https://www.semanticscholar.org/paper/Mathematical-Model-for-Enhanced-Production-of-Using-Mohammed/c9f4af9e5f8fe747b1e7bcb30fba0501bb3bae0c.
    The Bacoside was found to be mixed growth-associated product with optimized specific growth rate(0.0721 d-1) in the batch cultivation of the Bacopa monnieri in this batch kinetic study. Brahmi (Bacopa monnieri, Family: Scrophulariaceae) widely distributed in wetland countries such as Asia, America, and Australia, is used as insanity, psychosis, insomnia, epilepsy, depression, Alzheimer’s disease, etc. Active compound saponins of this plant, Bacoside are widely used for boosting memory and combating stress. Memory enhancing properties of this compound had huge market demand as compared to its production. Callus was induced on MS basal media fortified with α-napthalenacetic acid (0.5 mg/L) and N6-benzylaminopurine (0.1 mg/L ). Every 2-week callus was subcultured in fresh MS medium. Optimized media was used for the maximal production of biomass/product. Suspension culture was established to find out biomass growth, substrate consumption, rate and product formation kinetics parameters. Biomass cultivation was, after that, established in the 500 ml Erlenmeyer flask containing, 1.3 g fresh callus in 200 ml MS callus inducing media. Growth-limiting substrate sucrose decreased till 30 days. Nitrate and phosphate concentration decreased continuously decreased till the 15th day. Biomass and secondary metabolite (Bacoside) increased throughout the process. After suspension culture, callus was dried at room temperature, and the amount of Bacoside was estimated in HPLC. In the 30 days of suspension culture study, 15.025 g/l fresh weight of callus was measured at the end of the process. Kinetic parameters were utilized further for development of a mathematical model to get optimium yield of secondary metabolite production. In this batch kinetic study, maximum Bacoside production (3.484 mg/l) was calculated. Rosenbrock optimization algorithm optimized the batch kinetic parameters. The Bacoside was found to be mixed growth-associated product with optimized specific growth rate(0.0721 d-1) in the batch cultivation of the Bacopa monnieri.
  46. Naik, Poornananda Madhava and Godbole, Manoj and Nagella, Praveen and Murthy, Hosakatte Niranjana, Influence of Different Media, Medium Strength and Carbon Sources on Adventitious Shoot Cultures and Production of Bacoside A in Bacopa Monnieri(L.), Ceylon Journal of Science, vol. 46, no. 4, pp. 97, December 2017. doi: 10.4038/cjs.v46i4.7472.
    Bacopa monnieri (L.) Wettst. (Plantaginaceae), commonly known as Brahmi, is used in Ayurveda as a medicinal herb, found throughout the Indian subcontinent in wet, damp and marshy areas. The herb yields pharmaceutically active compounds called bacosides. The current investigation was carried out to assess the potential for increasing biomass and the concentration of bacoside A from the in vitro regenerated shoots in different media (MS, B5, NN and N6), the strength of the MS medium (0.25, 0.50, 0.75, 1.0, 1.50 and 2.0X) and carbon source [sucrose, glucose, fructose, maltose, sucrose + glucose (1:1), sucrose + maltose (1:1), glucose + fructose (1:1), glucose + maltose (1:1), fructose + sucrose (1:1) and fructose + maltose (1:1)]. All the culture media were supplemented with 2 mg/L Kinetin (Kin) at pH 5.8. High performance liquid chromatography (HPLC) was used to determine the bacoside A content from the regenerated shoots. Optimum number of adventitious shoots (70.75 shoots per explant), fresh weight (2.344 g), dry weight (0.166 g) and bacoside A content (13.052 mg/g DW) were obtained from the cultures grown in the MS medium. Bacoside A production was highest in the shoots grown in the glucose + fructose combination (15.588 mg/g DW) among the different carbon sources tested. These findings suggest that B. monnieri culture requires full strength MS medium and a moderate level of sucrose. The present study is useful to obtain optimum production of biomass and bacoside A from the in vitro regenerated shoots of B. monnieri.
  47. Dixit, Himisha and Thakur, Ajay, In Vitro Propagation of a Medicinal Important Plant Bacopa Monnieri from Nodal Explants, pp. 4, 2017.
    Bacopa monnieri, a traditional Indian medicinal plant with high commercial importance, is used as a neurotonic, immuno modulator, adaptogen transquilizing, memory and learning enhancing, cerebral activator, anti-ulcer, antispasmodic, anti-asthmatic ayurvedic herb. The study was undertaken in order to elaborate the tissue culture technique for Brahmi (Bacopa monnieri). In the present study nodal explants were taken for in vitro propagation. For sterilization, 0.01\% HgCl2 was used as the sterilant. The explants gave best results when treated with the sterilant for 5 minutes. To induce bud-break, nodal segments were inoculated onto MS media supplemented with 1mg/l BAP. Basal MS medium without any plant growth hormone was used as control. Further multiplication was obtained on MS medium + BAP (1.0 mg/l) +IAA (0.5 mg/l). The shoots were rooted and best rooting was observed by IBA when incorporated in MS at different concentrations (0.1 - 0.3 mg/l) and 0.2 mg/l gave good results. After 30 days about 2.2 cm length was observed. Almost 70\% of the rooted shoots survived during hardening.
  48. Krishnaveni, M., In Vitro Micropropagation of Bacopa Monnieri, Its Antimicrobial Activity, 2017. doi: 10.20959/WJPR20178-8987.
    The micropropagation protocol of Bacopa monnieri for surface sterilization, suitable media for aseptic culture initiation, establishment and multiplication, rooting, optimization of medium for callus induction, analyse phytochemical contents, antimicrobial activity was standardized. Bacopa monnieri (BM) is a small, creeping, glabrous, succulent herb possessing antioxidant, antimicrobial properties and widely used by ayurvedic system of medicine since olden days. Hence, it was decided to standardize micropropagation protocol of Bacopa monnieri for surface sterilization, suitable media for aseptic culture initiation, establishment and multiplication, rooting, optimization of medium for callus induction, analyse phytochemical contents, antimicrobial activity. The best medium observed for initiation and development in terms of bud breakage percentage, cluster formation and maximum shoot length were BM1 (MS+0.5 mg/l BAP + 0.5 mg/l NAA) with shoot length of 4.2 cm; 80\% bud breakage and 65\% cluster formation and BM3 (MS+4 mg/l BAP + 0.4 mg/l NAA) with shoot length of 3.8 cm; 100\% bud breakage but cluster formation rate was low but this rate increased further in the multiplication stage when compared to BM4 (0.5 mg/l BAP + 0.5 mg/l.), BM5 (MS+ 1.0mg/l BAP + 1.0mg/l) medium. Multiplication stage was good with BM3 (4 mg/l BAP + 0.4 mg/l NAA) and BM1 (0.5 mg/l BAP + 0.5 mg/l NAA) continued to give best results for the average shoot length of 5.3 cm in BM3 and 4.60 cm in BM1. Extracts was found to have antimicrobial activity on K. pneumoniae, P. aerugenosa, B.stearothermophilus, B. substilis and S. aureus with zones of inhibitions ranging from 10-26mm.
  49. Kharde, Ashok and Chavan, Nilesh and Chandre, Mahesh and Haribhau, Autade and MB, Khetmalas, In Vitro Enhancement of Bacoside in Brahmi (Bacopa Monnieri) Using Colchicine, Journal of Plant Biochemistry \& Physiology, vol. 5, pp. 2--6, January 2017. doi: 10.4172/2329-9029.1000172.
    Present investigation on in vitro effects of two levels of colchicine (0.1\% and 0.2\%) for 1, 2, 3, 4 and 5 hours treatment on leaf explants of Brahmi (Bacopa monnieri) collected from Paithan (Aurangabad) was conducted with a view to induce somaclonal variations and enhancement in the bacoside content in regenerated plants. Bramhi plants regenerated through previously treated leaf explants with colchicine showed morphological variations in leaf shape, number of leaves per node, leaf arrangement, etc. and enhance the bacoside content. 0.2\% colchicine treatment for 5 hours showed maximum variations in number of leaves per node under in vitro conditions; however, these variations were not continued after subculture. While the 0.1\% colchicine treatment for 2 hours significantly increase the bacoside content up to 0.72\% i.e., more than four folds to the in vitro regenerated Brahmi plants without colchicine treatments.
  50. Haque, Sk Moquammel and Chakraborty, Avijit and Dey, Diganta and Mukherjee, Swapna and Nayak, Sanghamitra and Ghosh, Biswajit, Improved Micropropagation of Bacopa Monnieri (L.) Wettst. (Plantaginaceae) and Antimicrobial Activity of in Vitro and Ex Vitro Raised Plants against Multidrug-Resistant Clinical Isolates of Urinary Tract Infecting (UTI) and Respiratory Tract Infecting (RTI) Bacteria, Clinical Phytoscience, vol. 3, no. 1, pp. 17, September 2017. doi: 10.1186/s40816-017-0055-6.
    Nowadays the multidrug-resistant (MDR) bacterial pathogens are a major concern of the medical science. Medicinal plants may be considered as new sources for producing antibacterial agents. The present study aimed to standardize an improved method for micropropagation and in vitro biomass production of Bacopa monnieri. Second aim is to evaluate the antimicrobial potency of in vitro cultured and ex vitro field grown micropropagated plants against different MDR clinical isolates of human urinary tract infecting (UTI) and respiratory tract infecting (RTI) pathogens.
  51. Sharma, Neelam and Singh, Rakesh and Pandey, Ruchira and Kaushik, Nutan, Genetic and Biochemical Stability Assessment of Plants Regenerated from Cryopreserved Shoot Tips of a Commercially Valuable Medicinal Herb Bacopa Monnieri (L.) Wettst, In Vitro Cellular \& Developmental Biology - Plant, vol. 53, no. 4, pp. 346--351, August 2017. doi: 10.1007/s11627-017-9826-5.
    Shoot tips from four accessions (IC249250, IC 426442, IC 375976, and IC468878) of Bacopa monnieri (L.) Wettst., a commercially valuable memory revitalizing medicinal plant, were cryopreserved using a vitrification technique. Depending on the genotype, 0 to 20\% plant regeneration without intermediary callus was achieved from cryopreserved shoot tips. Genetic stability of plants derived from cryopreserved shoot tips was assessed using biochemical and molecular markers. The regenerated plants from non-frozen controls and cryopreserved shoot tips exhibited morphological similarity to respective parental material when transferred to soil. On the basis of ten random amplified polymorphic DNA (RAPD) and bacoside A content using HPLC analysis, no significant reproducible variation was observed between the controls and in vitro-cryopreserved plants. Thus, after cryopreservation treatment, the regenerated plants exhibited molecular and biochemical genetic stability.
  52. Meenashree, B. and Kathiravan, G. and Srinivasan, K. and Rajangam, B., Effect of Plant Hormones and Media Composition on Browning and Growth of Bacopa Monnieri Callus Cultures, Research Journal of Pharmacy and Technology, vol. 10, no. 2, pp. 497, 2017. doi: 10.5958/0974-360X.2017.00099.3.
  53. L and Mehta, A., Effect of Plant Growth Regulators on Callus Multiplication and in Vitro Plant Regeneration in Bacopa Monnieri L, International Journal of Medicinal Plants Research, vol. 6, no. 5, pp. 337--345, May 2017. url: https://www.internationalscholarsjournals.com/abstract/effect-of-plant-growth-regulators-on-callus-multiplication-and-in-vitro-plant-regeneration-in-bacopa-monnieri-l-57879.html.
    Current investigation deals with the tissue culture studies on Bacopa monnieri which has very high morphogenic potential. Its leaf explants respond very readily to treatment with auxins and cytokinins. Indefinite number of plantlets were regenerated from leaf margin, without intervening callus on Murashige \& Skoog (MS) medium supplemented with indole-3acetic acid (IAA) and kinetin (KN). Green embryogenic callus with indefinite number of micro shoots were developed on naphthalene acetic acid (NAA) supplemented medium when 2,4-dichlorophenoxyacetic acid (2,4-D) grown stem-derived callus was sub-cultured. During present investigation, stem (internode, node) explants exhibited callus induction, whereas, leaf explant showed a tendency to regenerate shoot and/or roots. In vitro grown plantlets were transferred to polypots containing sterile cocopeat and then series of steps were carried out for acclimatization. Survival rate was 100\% when acclimatized plantlets were transferred to soil. Micropropagated plantlets were without any morphological abnormalities.
  54. Srivastava, Preeti and Tiwari, Kavindra Nath and Srivastava, Gaurava, Effect of Different Carbon Sources on in Vitro Regeneration of Brahmi Bacopa Monnieri (L.) An Important Memory Vitalizer, Journal of Medicinal Plants Studies, vol. 5, no. 3, pp. 202--208, 2017. url: https://www.plantsjournal.com/archives/?year=2017&vol=5&issue=3&part=C&ArticleId=623.
    Brahmi (Bacopa monnieri) is an important medhyarasayan plant and it is widely used in Indian traditional system of medicine for the improvement of memory/brain function. This medicinal herb is exploited from the wild population all over world so there is need to develop the in vitro protocol for the mass multiplication under various conditions and its conservation. The International Union for Conservation of Nature and Natural Resources (IUCN) has a long time ago listed B. monnieri as a threatened species. MS medium containing 8.9 μM BA was found most suitable for induction of high frequency shoot buds from different explants e.g., leaf and node. We had evaluated different concentration range (0.5-10\%) of different carbon sources (sucrose, mannitol and glucose) for enhancement of this regeneration event. It was observed that MS media modified with 5\% sucrose was found most effective for shoot bud induction and shoot regeneration from nodal explants and it ranges 22.6 shoots/explants and for leaf explant 3\% sucrose was found effective with 20.6 shoots/explant. These shoots were further elongated on MS basal media without growth regulator. For getting maximum rooting these elongated shoots were transferred on MS media containing 4.9 μM IBA. The plantlets after acclimatization transferred into field with 90\% survival rate. Thus the present study could be utilized for better multiplication and conservation of this important threatened species.
  55. Anju, Varshney and Naresh, Chandra and Avinash, Patil, Anatomical Markers and Phytochemical Study of Different Plant Parts of Bacopa Monnieri (L.) Wettst., International Journal of Life Sciences, vol. 5, no. 3, pp. 379--386, 2017. url: https://oaji.net/articles/2017/736-1505841184.pdf.
    Bacopa monnieri (L.) Wettst. (Scrophulariaceae) commonly known as “brahmi”, has been used in the traditional systems of medicines as "Medhya Rasayana" as it increases mental clarity and brain stimulating action. It is also being used as anti-inflammatory, analgesic, antipyretic, sedative, antioxidant and anti-cancerous agent. The present study was carried out to provide characteristic anatomical markers for root, stem and leaf, along with this study preliminary photochemical analysis was also carried out for the presence of major secondary metabolites which can act as the indicator of bioactivity of the different plant parts of Bacopa monnieri (L.) Wettst. The distinguishing features for root and stem are simple starch grains, xylem vessel with spiral thickenings, pitted tracheids and for leaf are subepidermal foliar idioblast, sessile, multicellular trichome and prismatic calcium oxalate crystals. Presence of two types of stomata i.e. anomocytic and diacyitc in the same field of vision under 45X magnification, are seen in the surface preparation as well as in the powder microscopy of Bacopa monnieri leaf. This is the major observation of the study. All these distinguishing features can be used as anatomical markers for correct indentification of root, stem and leaf of the plant. Preliminary phytochemical analysis revealed the presence of alkaloids, saponins, flavonoids, tannins, carbohydrates, proteins and anthroquinones in different plant parts of Bacopa monnieri while quantitative estimation showed that alkaloids (mg /g dry weight) in root, stem and leaf were found to be 1.67 ± 0.577, 47.00 ± 0.81 and 53.07 ± 2.08 respectively; saponins (mg /g dry weight) in root, stem and leaf were 7.50 ± 0.71, 23.33 ± 1.53 and 57.00 ± 2.65 respectively and flavonoids (mg /g dry weight) in stem and leaf were 25.33 ± 1.53 and 27.67 ± 2.08 respectively. Thus the findings of the present study will provide referential pharmaco - botanical information for correct identification and authentication of the crude drug and its plant parts.
  56. Sharma, Neelam and Singh, Rakesh and Pandey, Ruchira, In Vitro Propagation and Conservation of Bacopa Monnieri L., pp. 153--171, 2016. doi: 10.1007/978-1-4939-3332-7_11.
    Bacopa monnieri L. (common name brahmi) is a traditional and renowned Indian medicinal plant with high commercial value for its memory revitalizer potential. Demand for this herb has further escalated due to popularization of various brahmi-based drugs coupled with reported anticancer property. Insufficient seed availability and problems associated with seed propagation including short seed viability are the major constraints of seed conservation in the gene banks. In vitro clonal propagation, a prerequisite for in vitro conservation by enhanced axillary branching was standardized. We have developed a simple, single step protocol for in vitro establishment, propagation and medium-term conservation of B. monnieri. Single node explants, cultured on Murashige and Skoog’s medium supplemented with BA (0.2 mg/L), exhibited shoot proliferation without callus formation. Rooting was achieved on the same medium. The in vitro raised plants were successfully transferred to soil with \textasciitilde 80 \% survival. On the same medium, shoots could also be conserved for 12 months with high survival and genetic stability was maintained as revealed by molecular markers. The protocol optimized in the present study has been applied for culture establishment, shoot multiplication and medium-term conservation of several Bacopa germplasm, procured from different agro-ecological regions of India.
  57. Narwal, Mrridula Dangi, Exploitation of Plant Extracts (Stem and Root) of Micropropagated Bacopa Monnieri: Antimycotic Potential, SMART MOVES JOURNAL IJOSCIENCE, vol. 2, no. 9, December 2016. doi: 10.24113/ijoscience.v2i9.116.
    Bacopa monnieriis commonly called as brahmi or jal brahmi in India. Brahmi is a non-aromatic herb Brahmi is considered as the herb played a very important role in Ayurvedic medicine. It is found easily India, Australia, Europe, Africa, Asia. Bacopa monnieri (L) belongs to the family Scrophulariaceae is an amphibious plant of tropical and normally found growing on the banks of the rivers and lakes. It is commonly called as brahmi or jal brahmi in India. Brahmi is considered as the main rejuvenating herb played a very important role in Ayurvedic therapies. It also has anti-inflammatory, analgesic, antipyretic, epilepsy, anticancer, antioxidant activities and recently antimycotic preoperty has been reported. The micro propagation protocol of the medicinally important plantBacopa monnieriwas standardized using nodal segments as explants. They were surface sterilized with HgCl2 (0.1\%) for 3 minutes prior to inoculation on MS media supplemented with BAP (0.5- 2.5 mg/l); IAA (0.1-0.5 mg/l for shooting,1.0-1.5 mg/l for rooting); NAA (0.1- 0.5 mg/l for shooting, 1.0-1.5 mg/l for rooting). The best performance for shoot multiplication was showed in MS medium supplemented with 1.5 mg/l BAP + 0.5 mg/l IAA. In this combination the number of shoots per explant was 16 and average length of shoot 5.54 ± 0.54 cm. But when different concentrations of NAA were applied along with 1.5 mg/l BAP the number of shoots per explant was 14 and average shoot length was 3.46 ± 0.43 on media. For root induction, best rooting was observed with half strength of MS medium supplemented with IAA (1.0 mg/l). In this combination, it was observed that the number of roots was 12 and average root length of 2.80 ± 0.09. The present study is a stepping stone for in vitro production of required active principles ofBacopa monnieri.
  58. Wangdi, Kencho and Sarethy, Indira P., Evaluation of Micropropagation System of Bacopa Monnieri L. in Liquid Culture and Its Effect on Antioxidant Properties, Journal of Herbs, Spices \& Medicinal Plants, vol. 22, no. 1, pp. 69--80, January 2016. doi: 10.1080/10496475.2015.1020404.
    Nodal explants of Bacopa monnieri were cultured in a solid Murashige and Skoog (MS) medium containing 1 mg L−1 kinetin, transferred after 30 days to a liquid MS medium of the same composition in 100 mL and 250 mL Erlenmeyer flasks (with 10 mL and 50 mL medium, respectively) and Phyta Jars (with 10 mL medium) and subcultured every 7 d. Shoot doubling time was estimated and the total antioxidant capacity of aqueous extracts from liquid-cultured and acclimatized plants was studied. In the solid medium, 5 rooted shoots/explant were produced after 20 days, with shoot doubling time of 7.99 days; in liquid media, 36–75 rooted shoots were produced with shoot doubling time of 0.91–1.17 days. Aqueous extracts of acclimatized plants showed 10-fold higher total antioxidant capacity compared to in vitro liquid cultured plantlets.
  59. Khilwani, Barkha and Kaur, Amanpreet and Ranjan, Ritika and Kumar, Anil, Direct Somatic Embryogenesis and Encapsulation of Somatic Embryos for in Vitro Conservation of Bacopa Monnieri (L.) Wettst, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 127, no. 2, pp. 433--442, November 2016. doi: 10.1007/s11240-016-1067-5.
    Direct somatic embryogenesis and shoot organogenesis were achieved from leaf explants excised from microshoots of Bacopa monnieri cultured on Murashige and Skoog medium containing N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of explants differentiated somatic embryos and shoot buds on MS medium supplemented with 12.5~µM BA and 1~µM 2,4-D. The frequency of explants differentiating somatic embryos decreased with increasing concentration of 2,4-D. Light and scanning electron microscopy revealed direct differentiation of somatic embryos and shoot buds from explants, and various developmental stages of the somatic embryos were observed. Somatic embryos and apical shoot tips were encapsulated in sodium alginate gel to produce synthetic seeds. The storage of synthetic seeds produced by encapsulation was studied at 4 and 25\,°C (room temperature) for a period of 140~days. Encapsulated somatic embryos were found to retain viability after 140~days of storage at both temperatures, whereas encapsulated apical shoot buds failed to germinate even after 40~days when stored at 4\,°C. The viability of synthetic seeds was higher when stored at 25\,°C. All amplified markers scored by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) were monomorphic for all the plants produced from synthetic seeds following different periods of storage, thus establishing the clonal fidelity of propagated plantlets.
  60. Govindan, Pothiaraj and Ebenezer, Sam and Christdas, Ebenezer and Shakila, Harshavardhan, Comparative Analysis on the Effect of Seaweed Liquid Extracts and Commercial Plant Growth Regulators on in Vitro Propagation of Bacopa Monnieri, issn 2319-284, vol. 519, pp. 1--9, April 2016.
    Employment of commercial Plant Growth Regulators (PGRs) is inevitable in plant tissue culturing, but it makes the process costlier. As a natural substitute with well-known plant growth promoting activity, seaweeds act as a plentiful source of these regulators. In this study, the plant growth enhancing property of Seaweed Liquid Extracts (SLEs) from Gracilaria edulis and Sargassum wightii was compared with that of the commercial PGRs during the in vitro propagation of Bacopa monnieri, a plant of high medicinal value. 0.1mg/ml of Naphthalene Acetic acid (NAA) and 1.0mg/ml of Kinetin were used to compare the plant growth promoting activity of commercial PGRs with Seaweed Liquid Extracts. Supplementation of MS media with different proportion of SLEs from S. wightii and G. edulis significantly enhanced the growth of Bacopa monnieri. 30\% and 40\% liquid extracts from S. wightii and G. edulis respectively enhanced shoot and root proliferation and increased the survivability of the propagated plant. Our results demonstrated that seaweed liquid extracts are more effective in enhancing the in vitro propagation of B. monnieri than commercial PGRs. Hence, the seaweed extracts as a source of the plant growth regulators are suggested as a better alternative for the commercial PGRs used for in vitro propagation of Bacopa monnieri.
  61. Suman, S., Tissue Culture Studies on Bacopa Monnieri: A Review, undefined, 2015. url: https://www.semanticscholar.org/paper/Tissue-Culture-Studies-on-Bacopa-monnieri%3A-A-Review-Suman/384f37b05f5f8df11d252817daac17a8744942f4.
    This review highlights the recent development and achievements made for the multiple shoots were regenerated from different explants and the induction of multiple shoots from nodal segments were found highest in MS medium. Abstract: As the plants goes endangered due to overexploitation and anthropogenic activities. There is~ necessity to develop a technique to conserve them micropropagation is one of them, to conserve bacopa a high economic value contained medicinal plant different tissue culture modes are used the growth agents were used at different concentration and high growth of regeneration was found. This review highlights the recent development and achievements made for the multiple shoots were regenerated from different explants. As a result the induction of multiple shoots from nodal segments were found highest in MS medium with 0.5 mg/l BAP + 2.0 mg/l Kn and 0.5 mg/l Kn+1.0 mg/l BAP. ~Highest rooting was recorded on MS medium supplemented with 2.0 mg/l IBA. ~In leaf petiole best growth was observed in MS medium supplemented with 0.25 mg/l 2, 4-D+ 0.5 mg/l Kn and 0.25 mg/l 2,4-D+ 0.1mg/l BAP. Key words: micropropagation, explants, propagule, MS medium.
  62. Sudheer, W. N. and Thiruvengadam, Muthu and Nagella, Praveen, A Comprehensive Review on Tissue Culture Studies and Secondary Metabolite Production in Bacopa Monnieri L. Pennell: A Nootropic Plant, Critical Reviews in Biotechnology, pp. 1--15, July 2022. doi: 10.1080/07388551.2022.2085544.
    Bacopa monnieri L. Pennell, commonly known as Brahmi, is an important medicinal plant that belongs to the family Plantaginaceae. Brahmi is rich in innumerable bioactive secondary metabolites, especially bacosides that can be employed to reduce many health issues. This plant is used as a neuro-tonic and treatment for mental health, depression, and cognitive performance. Brahmi is also known for its antioxidant, anti-inflammatory, and anti-hepatotoxic activities. There is a huge demand for its raw materials, particularly for the extraction of bioactive molecules. The conventional mode of propagation could not meet the required commercial demand. To overcome this, biotechnological approaches, such as plant tissue culture techniques have been established for the production of important secondary metabolites through various culture techniques, such as callus and cell suspension cultures and organ cultures, to allow for rapid propagation and conservation of medicinally important plants with increased production of bioactive compounds. It has been found that a bioreactor-based technology can also enhance the multiplication rate of cell and organ cultures for commercial propagation of medicinally important bioactive molecules. The present review focuses on the propagation and production of bacoside A by cell and organ cultures of Bacopa monnieri, a nootropic plant. The review also focuses on the biosynthesis of bacoside A, different elicitation strategies, and the over-expression of genes for the production of bacoside-A. It also identifies research gaps that need to be addressed in future studies for the sustainable production of bioactive molecules from B. monnieri.
  63. Mishra, S. and Tiwari, K. N. and Mishra, A., Micropropagation and Comparative Phytochemical, Antioxidant Study of BacopaMonnieri (L.) Pennell., undefined, 2015. url: https://www.semanticscholar.org/paper/Micropropagation-and-Comparative-Phytochemical%2C-of-Mishra-Tiwari/6bc4d1c78528def04aabaa812eb39ab2e370cf35.
    The protocols for tissue culture has been developed for regeneration of the BM plant from small part (nodal segment) and the regenerated plant having the similar potential in terms of phytoconstituents and antioxidant activity. Bacopa monnieri an important medicinal plant of Scorphulariaceae family and is distributed in the wet and marshy lands throughout India, Nepal, Sri Lanka, China, Taiwan, and Hawaii. It shows the presence of various phytoconstituents namely, alkaloids, saponins, phenolics and flavonoids. These phytoconstituents are responsible for anti oxidant and various other activity of the plant. The protocols for tissue culture has been developed for regeneration of the BM plant from small part (nodal segment). The regenerated plant having the similar potential in terms of phytoconstituents and antioxidant activity. The In vitro (leaf and stem) regenerated plants have slightly less antioxidant activity as compared to the In vivo but comparable. The protocols (MS+BAP5µg/ml) may used for the clonal regeneration of plant on large scale through which it can be protected and preserved for long duration in small space. A good response was observed in a soil ratio 3:2:1
  64. Largia, Muthiah Joe Virgin and Pothiraj, Govindan and Shilpha, Jayabalan and Ramesh, Manikandan, Methyl Jasmonate and Salicylic Acid Synergism Enhances Bacoside A Content in Shoot Cultures of Bacopa Monnieri (L.), Plant Cell, Tissue and Organ Culture (PCTOC), vol. 122, no. 1, pp. 9--20, July 2015. doi: 10.1007/s11240-015-0745-z.
    This study has been carried out with the aim of enhancing the production of bacoside A in plant cultures of Bacopa monnieri through the process of elicitation. Optimisation of suitable concentrations of plant growth regulators and harvest time has been accomplished for the highest biomass production and bacoside A accumulation. Plant cultures were elicited with methyl jasmonate (MJ) and salicylic acid (SA) separately and in combinations in different concentrations for a period of 1–4~weeks. Three weeks of elicitation with 50~µM MJ or 50~µM SA produced about two and threefold higher quantity of bacoside A respectively when compared to unelicited controls. Highest accumulation was noticed in a combination of 25~µM MJ~+~25~µM SA treated for 3~weeks duration. It has produced about 59.18~mg/g DW (dry weight) of bacoside A3 (3.5-fold higher), 95.30~mg/g DW of Bacopaside II (5.2-fold higher), 46.11~mg/g DW of Jujubogenin isomer of Bacopasaponin C (4.8-fold higher) and 68.16~mg/g DW of Bacopasaponin C (5.9-fold higher), altogether accounting for the production of 269.71~mg/g DW (almost fivefold higher) of bacoside A. Hence the protocol formulated provides an efficient alternative for mass production of whole plants with increased metabolite content.
  65. Gantait, Saikat and Kundu, Suprabuddha and Ali, Nasim, Influence of Encapsulating Agent and Matr Ix Levels on Synseed Production of Bacopa Monnieri (L.) Pennell, Medicinal Plants - International Journal of Phytomedicines and Related Industries, vol. 7, no. 3, pp. 182, 2015. doi: 10.5958/0975-6892.2015.00026.X.
    The present study reports on the influence of sodium alginate and calcium chloride (CaCl2.H20) levels on proficient synthetic seed (synseed) formation of Bacopa monnieri (L.) Pennell. In vitro nodal segments (\textasciitilde 4 mm) were employed for encapsulation wherein aliquots of 1.5-4\% (w/v) (i.e. 1.5, 2, 2.5, 3, 3.5, and 4\%) sodium alginate containing individual nodal segments were dropped in either 75 mM or 100 mM calcium chloride. Uniform spherical beads were obtained using 2.5\% sodium alginate and 75 mM calcium chloride. Earliest conversion (into plantlet with shoot and root) of synseeds derived from optimized sodium alginate and calcium chloride level, was achieved on half-strength Murashige and Skoog semisolid regrowth medium. The synseeds displayed distinctive characters in their response to variable levels of sodium alginate and calcium chloride. The comparatively lower (1.5 and 2\%) or higher (3, 3.5 and 4\%) sodium alginate levels accompanied with more concentrated (100 mM) calcium chloride resulted either in squashy, fragile or harder and tail-forming beads, respectively.
  66. Sharma, Munish and Ahuja, Ashok and Gupta, Rajinder and Mallubhotla, Sharada, Enhanced Bacoside Production in Shoot Cultures of Bacopa Monnieri under the Influence of Abiotic Elicitors, Natural Product Research, vol. 29, no. 8, pp. 745--749, April 2015. doi: 10.1080/14786419.2014.986657.
    The effect of different abiotic elicitors [jasmonic acid, copper sulphate (CuSO4) and salicyclic acid] at varying concentrations on the stimulation of biomass and bacoside production in in vitro Bacopa monnieri shoot culture was studied. A systematic study conducted over a period of 35 days indicated that the maximum bacoside production (6.74 mg g− 1 dry weight (DW)) was obtained after a lag of 7 days and thereafter, the content decreased gradually to again increase at 28 days (5.91 mg g− 1 DW). Therefore, elicitation experiments were carried out over a period of 3, 6 and 9 days. The shoot cultures treated with 45 mg L− 1 of CuSO4 exhibited the highest bacoside content of 8.73 mg g− 1 DW (∼1.42-fold higher) than in control cultures (6.14 mg g− 1 DW). This study indicates the effectiveness of abiotic elicitation on bacoside production in in vitro shoot cultures of this medicinally important herb known for its memory-enhancing properties.
  67. Paul, Pijush and Sarkar, Sayantika and Jha, Sumita, Effects Associated with Insertion of Cryptogein Gene Utilizing Ri and Ti Plasmids on Morphology and Secondary Metabolites Are Stable in Bacopa Monnieri-Transformed Plants Grown in Vitro and Ex Vitro, Plant Biotechnology Reports, vol. 9, no. 4, pp. 231--245, July 2015. doi: 10.1007/s11816-015-0360-9.
    Bacopa monnieri (Linn.) Wettst. (family Scrophulariaceae), a therapeutically important perennial herb, was transformed to study the stability of the effects associated with the insertion of a gene encoding β-cryptogein, a proteinaceous elicitor, either alone or in addition to the T-DNA genes in long-term in vitro culture as well as after transfer to the greenhouse. Plant lines BmAr-IXcrypt obtained via transformation by LBA 9402-crypt (pRi1855+pBin19++crypt) showed integration and expression of rolA, rolB, rolC, and rolD genes after 4~years of maintenance in vitro. The plant line BmAt-ncrypt obtained via transformation with A. tumefaciens strain LBA4404-crypt (harboring pBin19++crypt) as well as plant line BmAr-IXcrypt showed stable integration and expression of nptII gene and crypt gene even after transfer to greenhouse. The clones of Ri-crypt-transformed plants differed morphologically from Ri-transformed plants in that the typical “hairy root syndrome” was not observed in plants expressing crypt gene in the presence of rol genes. Except plant line BmAr-IXcrypt, none of the other transgenic plant lines showed any alteration in floral morphology and onset of flowering. The crypt-transformed plant lines (BmAr-IXcrypt and BmAt-ncrypt), maintained under long-term culture, showed significantly higher (p~≤~0.05) amount of bacoside production in vitro (1.66- to 2.05-fold, with respect to non-transformed plants). In addition, accumulation of all the four bacosides was significantly higher (p~≤~0.05) in crypt-transformed plants as compared to non-transformed plants grown for 1~year in greenhouse. Thus, the crypt-transformed plant lines of B. monnieri may be considered as the potential source for sustainable bioproduction of bacosides.
  68. Largia, Muthiah Joe Virgin and Shilpha, Jayabalan and Pothiraj, Govindan and Ramesh, Manikandan, Analysis of Nuclear DNA Content, Genetic Stability, Bacoside A Quantity and Antioxidant Potential of Long Term in Vitro Grown Germplasm Lines of Bacopa Monnieri (L.), Plant Cell, Tissue and Organ Culture (PCTOC), vol. 120, no. 1, pp. 399--406, January 2015. doi: 10.1007/s11240-014-0602-5.
    The true-to-type nature of shoot cultures of Bacopa monnieri maintained in vitro for 5~years has been confirmed by means of flow cytometry (FCM), inter simple sequence repeat (ISSR), directed amplification of mini satellite region DNA (DAMD) analysis and chemical profiling. The nuclear DNA content of B. monnieri has been estimated as 1.86~pg/2C based on the FCM analysis of nuclei from young leaves. It has also detected no significant variation in the nuclear DNA contents between long term maintained germplasm lines and field grown plants. ISSR and DAMD analysis produced about 93 amplification products and all of them were found to be monomorphic among the field grown and seven germplasm lines. Even though morphological growth parameters varied slightly between the lines, the bioactive compound production capacity has been retained. The mean amount of Bacoside A accumulated by the germplasm lines has been quantified as 25.02~mg~g−1 dry weight (DW) using high performance liquid chromatography analysis which is comparable with the quantity of Bacoside A accumulated by the field grown plants (26~mg~g−1 DW). The total phenolics (folin–ciocalteau method; expressed in milligram Gallic acid equivalents per gram dry weight of the sample) and the flavonoids content (aluminium chloride method; expressed in milligram Quercetin equivalents per gram dry weight of the sample) of the germplasm lines (53.50–56.62~mg GAE/g DW; 273.51–277.16~mg QRE/g DW) were found to be slightly higher than the field grown plants (46.66~mg/g GAE; 256.65~mg/g QRE) respectively. Further, the antioxidant potential of the methanolic whole plant extracts determined by means of (1,1-diphenyl-2-picrylhydrazyl) DPPH assay revealed that the germplasm lines showed little higher antioxidant potential with IC50 values ranging from 39.51 to 43.71~µg~g−1 DW when compared to the IC50 value of field grown (45.68~µg~g−1 DW). Hence the protocol devised using axillary bud culturing can be utilized effectively for the long term maintenance of aseptic clones with high genetic and chemical fidelity. The approach tackles the increasing market demand through the continuous availability of high quality plant materials all round the year besides preserving the lines under monitored in vitro conditions.
  69. Behera, Shashikanta and Nayak, Nibedita and Hota, Shasmita and Barik, D. and Naik, S. K., An Efficient Micropropagation Protocol of Bacopa Monnieri (L.) Pennell through Two-Stage Culture of Nodal Segments and Ex Vitro Acclimatization -, 2015. doi: 10.7324/jabb.2015.3304.
    Cent per cent acclimatization of healthy in vitro regenerated shoots was obtained by ex vitro rooting in plastic pots containing garden soil, which saves time and tissue culture cost by abolishing the in vitro rooting step. An efficient plant regeneration protocol through two-stage culture of nodal segment has been reported here for a valuable medicinal plant, Bacopa monnieri. Multiple shoots with large number of shoot buds were induced from nodal explant on Murashige and Skoog’s (1962) (MS) medium fortified with (1.0-5.0 mg L-1) N6- benzyladenine (BA). Of the different concentrations of BA tested, 3.0 mg L-1 BA offered the best result, where the nodes swelled and an average of 6.5 shoots with numerous buds/nodes were recorded. The swollen nodes, cultured on MS + 3.0 mg L-1 BA, were sub-cultured either on MS or MS supplemented with 1.0 mg L-1 GA3 medium. The best result (114.2 shoots/ node) with an average shoot length of 6.4 cm was observed on MS media supplemented with 1.0 mg L-1 GA3. Cent per cent acclimatization of healthy in vitro regenerated shoots was obtained by ex vitro rooting in plastic pots containing garden soil, which saves time and tissue culture cost by abolishing the in vitro rooting step.
  70. Bansal, Mahima and Kumar, Anil and Sudhakara Reddy, M., Influence of Agrobacterium Rhizogenes Strains on Hairy Root Induction and ‘Bacoside A’ Production from Bacopa Monnieri (L.) Wettst., Acta Physiologiae Plantarum, vol. 36, no. 10, pp. 2793--2801, October 2014. doi: 10.1007/s11738-014-1650-5.
    Stable lines of hairy roots were established from leaf explants of Bacopa monnieri using different strains (A4, R1000, SA79, MTCC 532 and MTCC 2364) of Agrobacterium rhizogenes. The efficiency of hairy roots induction of these strains varied significantly and the maximum transformation frequency (75~\%) was observed in case of strain SA79 using leaf explants followed by internode (55~\%) in the presence of acetosyringone. Different parameters such as cell density of Agrobacterium suspension, co-cultivation period and infection time influenced the root induction frequency. Maximum frequency of root induction was obtained with bacterial density of 0.6 OD600, 2~days of co-cultivation period and 10~min of infection time. Integration of T-DNA in the genome of hairy roots was confirmed by PCR amplification of rolB gene. Elimination of Agrobacterium from the established root cultures was ascertained by amplifying the DNA fragment specific to 16S rDNA and virD gene. All lines of hairy roots except strain A4 induced showed higher growth rate and accumulated higher levels of ‘bacoside A’ than the untransformed roots. Maximum biomass accumulation (6.8~g~l−1) and ‘bacoside A’ content (10.02~mg~g−1 DW) were recorded in case of the hairy root line induced by strain MTCC 2364.
  71. Kumari, Ritika and Priyadarshni, M. and Anjali, Kumari Anjali Kumari, In Vitro Mass Multiplication of Bacopa Monnieri (l.) an Endangered and Valuable Medicinal Herb, undefined, 2014. url: https://www.semanticscholar.org/paper/IN-VITRO-MASS-MULTIPLICATION-OF-Bacopa-monnieri-AN-Kumari-Priyadarshni/8b992cac2d7a880a071ea5444c5e50f25e9a930f.
    Bacopa monneiri (L.) an important medicinal herb, commonly used as memory vitalizer has been used for mass multiplication through culture of nodal and apical shoot apex in MS basal medium supplemented with various concentrations of 6-Benzyle amino purine (BAP), Kinetin, either alone or in combinations with IBA. Bacopa monneiri (L.) an important medicinal herb, commonly used as memory vitalizer has been used for mass multiplication through culture of nodal and apical shoot apex in MS (Murashige and Skoog, 1962) basal medium supplemented with various concentrations of 6-Benzyle amino purine (BAP), Kinetin, either alone or in combinations with IBA. Both BAP and KN at 0.5 mg/l concentration were found most suitable Cytokinin with respect to multiple shoot induction. At this concentration, the mean number of shoots on nodal explants was 18.0 where as it was 15.0 in shoot tip explants on subculture in the same composition of the medium. However, in case of KN,at the same concentration the mean number was 16 in the nodal explants and 12 in the shoot tip explants.When both the Cytokinin were used together, no promising result was obtained. This was also true for the supplementation of IBA. Well-developed shoots raised in vitro were excised and used for rooting in MS basal medium supplemented with various concentration of IBA.95\% explants rooted in the medium supplemented with 0.25 mg/l IBA where the mean number of roots was 12.0 and average length 5.6 cm after 4 weeks. Well rooted plants were carefully taken out from the medium and washed properly with glass distilled water to remove the agar adhere with the roots. They were planted in the rooting pot containing 1:1:1 autoclaved vermiculite sand and soil. Humidity of the chamber (polybag) was maintained through water soaked sponge platform. Half strength liquid MS medium was used for irrigation on alternate day.82\% plantlets survived in this condition while survival rate in the field was 72\%.
  72. Ahire, M. L. and Laxmi, S. and Walunj, P. R. and Kishor, P. B. Kavi and Nikam, T. D., Effect of Potassium Chloride and Calcium Chloride Induced Stress on in Vitro Cultures of Bacopa Monnieri (L.) Pennell and Accumulation of Medicinally Important Bacoside A, Journal of Plant Biochemistry and Biotechnology, vol. 23, no. 4, pp. 366--378, October 2014. doi: 10.1007/s13562-013-0220-z.
    Growth, osmotic adjustment, antioxidant enzyme defense and principle medicinal component bacoside A was studied in in vitro raised shoots of Bacopa monnieri under different concentrations of KCl and CaCl2 (0, 50, 100, 150 or 200~mM). Significant reduction was observed in shoot number per culture; shoot length, fresh weight, dry weight and tissue water content (TWC) when shoots were exposed to increasing KCl and CaCl2 concentrations (50–200~mM) as compared to control. Minimum damage to the membrane as assessed by malondialdehyde (MDA) content was noticed in control in contrast to sharp increase in KCl and CaCl2 stressed shoots. Higher amounts of free proline, glycine betaine and total soluble sugars (TSS) accumulated in KCl and CaCl2 exposed shoots compared to the controls. Among different concentrations of KCl and CaCl2, increasing concentration of CaCl2 showed more increase in osmolyte accumulation. Na+ content decreased with increasing concentrations of KCl and CaCl2. Accumulation of K+ increased significantly in KCl (50–100~mM) stressed shoots as compared to control, while it decreased in CaCl2 treated shoots indicating that it prevents the uptake of K+ ions. Ca2+ accumulation significantly increased with increasing concentrations of CaCl2 up to 150~mM but decreased at higher concentrations. Shoots treated with KCl and CaCl2 (0–100~mM) showed higher antioxidant enzyme (SOD, CAT, APX and GPX) activities but KCl suppressed the activities at higher concentrations. Accumulation of bacoside A was enhanced with an increase in KCl and CaCl2 concentration up to 100~mM. It appears from the data that accumulation of osmolytes, and elevated activities of antioxidant enzymes play an important role in osmotic adjustment in shoot cultures of Bacopa and the two salts tested have a positive effect on bacoside accumulation.
  73. Łojewski, Maciej and Muszyńska, Bożena and Smalec, Agata and Reczyński, Witold and Opoka, Włodzimierz and {Sułkowska-Ziaja}, Katarzyna, Development of Optimal Medium Content for Bioelements Accumulation in Bacopa Monnieri (L.) In Vitro Culture, Applied Biochemistry and Biotechnology, vol. 174, no. 4, pp. 1535--1547, October 2014. doi: 10.1007/s12010-014-1095-8.
    Bacopa monnieri is one of the most interesting plants from the Ayurveda system. The aims of present research were, basing on in vitro shoot culture of B. monnieri, to determine content and to evaluate the influence of physiologically important metabolites on the selected bioelements accumulation in biomass. The most significant increase in biomass production was observed in the culture medium enriched with 0.5~mg/L of anthranilic acid. In this medium also, the highest accumulation of Mg was noted. The highest concentration of iron was determined in B. monnieri in vitro culture enriched with 0.25~g/L of serine. The addition of l-tryptophan, magnesium sulfate, and zinc hydroaspartate caused only a small increase in the accumulation of copper in B. monnieri. Increase in Zn accumulation was obtained in biomass from in vitro culture of B. monnieri with the addition of magnesium sulfate and zinc hydroaspartate. In the case of Na, the maximum level of this element was in biomass from medium enriched with zinc hydroaspartate. Twofold increase in K concentration was obtained in biomass from cultures on medium with addition of serine and magnesium sulfate. The concentrations of Ca in biomass of all studied media were at the similar level.
  74. Koul, Anuja and Sharma, Arti and Gupta, Sofia and Mallubhotla, Sharada, Cost Effective Protocol for Micropropagation of Bacopa Monnieri Using Leaf Explants, International Journal of Science and Research, vol. 3, pp. 201--212, April 2014.
    Owing to the resurgence in herbal based therapies, plants which serve as raw materials are required in large quantities to meet their increasing demand. Additional efforts are employed through various conventional and non-conventional ways for propagation and conservation of medicinal plants. Bacopa monnieri (L.) Wettst (Family: Scrophulariaceae) is one such medicinal plant used in Indian traditional systems of cure for various diseases and thus it becomes necessary to conserve this plant species. An efficient, cost effective protocol has been standardized using leaves as explants for in vitro propagation of B. monnieri. Leaves of five different accessions of B. monnieri collected from various regions of North India were cultured on agar gelled Murashige \& Skoog (MS) \& Gamborg’s (B5) media without the addition of any expensive nutritional supplements (plant growth regulators, growth additives). De novo shoot initiation from leaf explants was observed after 20 days of inoculation accompanied by rooting in the form of a single tap root in all the accessions in both the media tested. Maximum shoots/explant (5.5±0.65), leaves/explant (17.9±0.75) and roots/explant (6.3±0.65) were obtained for leaves of Accession BM003 cultured on MS media. Through this study we conclude that MS media is superior over B5 media for in vitro shoot multiplication and plantlet regeneration of B. monnieri. The protocol can cut down the cost of micropropagation of this endangered herb and also minimize the excessive and reckless use of plant propagation explants (shoot tips, nodes).
  75. Talukdar, Abhijit, Biosynthesis of Total Bacosides in the Callus Culture of Bacopa Monnieri. L. Pennel from North-East India, undefined, 2014. url: https://www.semanticscholar.org/paper/Biosynthesis-of-Total-Bacosides-in-the-callus-of-L.-Talukdar/42598e993805c44b6be566fcc1f84d6b354db7c5.
    Bacopa monnieri, commonly and widely called as brahmi, belongs to the family Scrophulariaceae and is used as a nerve tonic in Ayurveda, Unani and Sidha system of medicines. Bacopa monnieri, commonly and widely called as brahmi, belongs to the family Scrophulariaceae. The plant is a small herb, prostrate, succlent, with branches spreading or ascending and rooting at the nodes. Leaves are ovate-oblong or spathulate and grows upto a height of about 18 cm. Flowers are white in color, companulate, solutary, axillary, short or long pedicilate and blossoms during the month of August and continues till the month of October. B. Monnieri mainly grows in the humid and watery areas and mainly distributed in damp and marshy tracts in the subtropical region upto 1200 m elevation. In India it is found to grow in almost all the states of the Northeast India Viz., Assam, Arunachal Pradesh, Nagaland, Manipur Mizoram Nagaland and Tripura and also in the states of West Bengal, Himachal Pradesh and other Himalayan regions. It requires a well drained, moist, sandy loamy soil, rich in organic matter at a temperature 30 C to 40 o C and plenty of rainfall. The whole herb is used in medicinal preparation in Ayurveda, Unani and Sidha system of medicines. It is used as a nerve tonic, ISSN: 2319-7706 Volume 3 Number 3 (2014) pp. 140-145 http://www.ijcmas.com
  76. Naik, P. and Patil, B. and Jaggal, L. and Jangid, V., Sciences The Effect of Subculture on the Bacoside A Content in Adventitious Shoot Cultures of Bacopa Monnieri (L), undefined, 2013. url: https://www.semanticscholar.org/paper/Sciences-The-Effect-of-Subculture-on-the-Bacoside-A-Naik-Patil/62c20bb59aa9129e5b7970b42225b4cc9fa95f50.
    There was gradual increase in the shoot number, biomass and bacoside content from parental culture and attained optimum at 6 th subculture, thereafter decrease in theshoot number. Brahmi (Bacopa monnieri) is an important medicinal plant mainly used for the improvement of intelligence, memory and revitalization of sensory organs. Recent investigations revealed that bacoside A is major chemical component shown to be responsible for memory facilitating action. In the present study the effect of subculture on adventitious shoot regeneration, biomass and bacoside a accumulation in Bacopa monnieri was carried out. The leaf explants were cultured on Murashige and skoog (MS) medium supplemented with 2.0 mg/L kinetin (Kin) and further they were subcultured in the same medium, and subcultured for every 2 months up to ten subcultures. There was gradual increase in the shoot number, biomass and bacoside content from parental culture and attained optimum at 6 th subculture, thereafter decrease in the shoot number. Optimum number of adventitious shoots (79.00 shoots per explant), fresh weight (2.800 g), dry weight (0.190 g) and highest production of bacoside content (13.750 mg/g DW) was recorded on 6 th subculture.
  77. Ghasolia, Bhagyashree and Shandilya, Deepti and Maheshwari, R., Multiple Shoot Regeneration of Bacopa Monnieri (L.) Using Cyanobacterial Media- A Novel Approach and Effect of Phytoregulators on in Vitro Micropropagation, undefined, 2013. url: https://www.semanticscholar.org/paper/Multiple-shoot-regeneration-of-Bacopa-monnieri-(L.)-Ghasolia-Shandilya/ad1a7c90554e0d0b22f7e2efc14bbf9a1684ffbf.
    This review highlights the recent development and n ovel achievements made for the multiple shoots regeneration of Bacopa monnieri in Cyanobacterial m edium as well as in MS medium. This review highlights the recent development and n ovel achievements made for the multiple shoots regeneration of Bacopa monnieri in Cyanobacterial m edium as well as in MS medium. Shootlets were regenerated from nodal explants of stem through aux iliary shoot proliferation. In Cyanobacterial culture medium Bacopa monnieri survived for long pe riod of time and multiple shoot initiation was observed in 2mg/l Kn. When Bacopa monnieri was inoc ulated in MS medium supplemented with different concentrations of PGR’s, maximum no. of s hoots were observed in 0.5 mg/l BAP + 2.0 mg/l Kn and 0.5 mg/l Kn+1.0 mg/l BAP.
  78. Sharma, Poojadevi and Yadav, Sheetal and Srivastava, Anshu and Shrivastava, Neeta, Methyl Jasmonate Mediates Upregulation of Bacoside A Production in Shoot Cultures of Bacopa Monnieri, Biotechnology Letters, vol. 35, no. 7, pp. 1121--1125, July 2013. doi: 10.1007/s10529-013-1178-6.
    Methyl jasmonate (MJ) enhances the production of a range of secondary metabolites including triterpenoid saponins in a variety of plant species. Here, it enhanced production of bacoside A, a valuable triterpenoid saponin having nootropic therapeutic activity in in vitro shoot cultures of Bacopa monnieri, the only known source of bacoside A. The highest yield was with 50~μM MJ giving 4.4~mg bacoside A/g dry wt; an 1.8-fold increase (compared to control) after 1~week.
  79. Asha, K. and Devi, A. and Dwivedi, N. and Nair, R., In Vitro Regeneration of Brahmi (Bacopa Monnieri (Linn.) Pennell) - an Important Medicinal Herb through Nodal Segment Culture, undefined, 2013. url: https://www.semanticscholar.org/paper/In-vitro-regeneration-of-Brahmi-(Bacopa-monnieri-an-Asha-Devi/621db498d0e4917edf2bf81c47b7ed04cfa93f49.
    An efficient and cost effective in vitro plant regeneration protocol through nodalsegment culture was achieved in the medicinally important herb Bacopa monnieri (L.)Pennell, the Memory Plus plant through axillary shoot proliferation in Murashige and Skooge medium augmented with varying concentrations of BAP. An efficient and cost effective in vitro plant regeneration protocol through nodalsegment culture was achieved in the medicinally important herb Bacopa monnieri (L.)Pennell, the Memory Plus plant through axillary shoot proliferation in Murashige and Skooge medium augmented with varying concentrations of 6-benzylaminopurine (BAP)1 - 5 mg/l. BAP at 2 mg/l was the most effective in multiple shoot induction and mean number of leaves, which gave an average of 17 shoots and 31.11 leaves, compared toother concentrations of the hormone tried in 35 days of culture. Regarding mean shoot length and number of nodes, basal MS giving 2.66 cm long shoots with 7.44 nodes is thebest. MS basal medium, even though not promoting shoot multiplication, gave highershoot length with elongated internodes. Healthy rooting of the in vitro developed shootswas achieved in half and full strength MS basal solid medium without the addition ofany hormones. The healthy and vigorous in vitro regenerated micro shoots wereseparated out and were hardened on transfer to plastic cups with sterile soil and sandand were successfully acclimatized ex vitro in pots with potting mixture under greenhouse conditions for 3 weeks. The survival rate was 100\% and the plants establishedwell in green house resembled the mother plants in habitat without any morphological variations. The very simple and cost effective protocol developed can be used to produceelite stable clones for en masse propagation for the large-scale cultivation of this very important medicinal herb.
  80. Kaur, J. and Nautiyal, K. and Pant, M., In Vitro Propagation of Bacopa Monnieri (L.) Wettst A Medicinally Priced Herb, undefined, 2013. url: https://www.semanticscholar.org/paper/In-vitro-propagation-of-Bacopa-monnieri-(L.)-Wettst-Kaur-Nautiyal/4827eb542089ae2b5168c570d079b15619396a78.
    Medium supplemented with 1.0 mg/l IBA proved to be most efficient for development of healthy root system and hardened plantlets were subsequently hardened, acclimatized and successfully established in field with 100\% survival rate. A B S T R A C T Tissue culture studies were performed on Bacopa monnieri (Brahmi). Nodal explants containing preformed axillary bud were used as the starting material. Surface sterilized explants were aseptically cultured on MS medium supplemented with different plant growth regulators. Best results of axillary bud induction were observed on medium containing 1.0 mg/l BAP wherein 100\% bud break was achieved. Optimal results of further shoot multiplication were also obtained on same media combination and shoots were periodically subcultured alternately on media containing 0.5 mg/l BAP and basal MS medium without any PGR. Half strength MS media containing different concentrations of auxins were used for in vitro rooting of shoots. Medium supplemented with 1.0 mg/l IBA proved to be most efficient for development of healthy root system. Rooted plantlets were subsequently hardened, acclimatized and successfully established in field with 100\% survival rate.
  81. Muthiah, Joe Virgin Largia and Shunmugiah, Karutha Pandian and Manikandan, Ramesh, Genetic Fidelity Assessment of Encapsulated in Vitro Tissues of Bacopa Monnieri after 6 Months of Storage by Using ISSR and RAPD Markers, Turkish Journal of Botany, vol. 37, no. 6, pp. 1008--1017, 2013. url: https://app.trdizin.gov.tr/makale/TVRVNE16RTJOZz09/genetic-fidelity-assessment-of-encapsulated-in-vitro-tissues-of-bacopa-monnieri-after-6-months-of-storage-by-using-issr-and-rapd-markers.
  82. Yusuf, A. and Nikhilesh, S. and Rao, P., Effects of Antioxidants and Gelling Agents on Regeneration , in Vitro Conservation and Genetic Stability of Bacopa Monnieri ( L . ), International Journal of Ayurvedic and Herbal Medicine, vol. 1, no. 3, pp. 51--67, 2013. url: https://www.semanticscholar.org/paper/Effects-of-antioxidants-and-gelling-agents-on-%2C-in-Yusuf-Nikhilesh/418032ab30b5fc630d7cfbe3c22b3f95dc9c675f.
    Shoot regeneration of Bacopa monnieri was achieved, on Murashige and Skoog (MS) medium supplemented with 4.4 μM benzyl aminopurine (BA) from different explants. Nodal explants initiated 75-80 shoots, leaf explants 55-65 shoots and internodal explants 50-55 shoots within 40 days of culture. The initiated shoots were multiplied in MS medium containing 2.2 μM BA. Ascorbic acid (50 mg/l) and citric acid (25 mg/l ) increased the number of shoots and conservation period of the cultures. Isabgol, when used as gelling agent provided consistent multiplication of cultures compared to microbiology grade agar and cultures were conserved in medium containing 3\% (w/v) isabgol for a period of 190 days as against an average of 120 days in 1\% (w/v) agar containing media. The conservation period was increased to a period of 200 days in medium gelled with 3\% (w/v) isabgol containing 50 mg/l ascorbic acid and 25 mg/l citric acid and more than 80\% of the cultures survived in this medium. Rooting was achieved ex vitro by culturing the isolated shoots in autoclaved soilrite moistened with 1/10 strength MS salts. Rooted plantlets were established in soil. Genetic stability assessment with random amplified polymorphic DNA (RAPD) primers showed no variation in the banding pattern of the cultures grown in different concentrations of growth regulators, agar or isabgol and additives containing media. These studies provide a feasible in vitro multiplication and conservation method and genetic stability assessment of B. monnieri cultures.
  83. Ahire, M. L. and Walunj, P. R. and Kishor, P. B. K. and Nikam, T. D., Effect of Sodium Chloride-Induced Stress on Growth, Proline, Glycine Betaine Accumulation, Antioxidative Defence and Bacoside A Content in in Vitro Regenerated Shoots of Bacopa Monnieri (L.) Pennell, Acta Physiologiae Plantarum, vol. 35, no. 6, pp. 1943--1953, June 2013. doi: 10.1007/s11738-013-1233-x.
    Growth, osmotic adjustment, antioxidant enzyme defense and the principle medicinal component bacoside A were studied in the in vitro raised shoot cultures of Bacopa monnieri, a known medicinal plant, under different concentrations of NaCl [0.0 (control), 50, 100, 150 or 200~mM]. A sharp increase in Na+ content was observed at 50~mM NaCl level and it was about 6.4-fold higher when compared with control. While Na+ content increased in the shoots with increasing levels of NaCl in the medium, both K+ and Ca2+ concentrations decreased. Significant reduction was observed in shoot number per culture; shoot length, fresh weight (FW), dry weight (DW) and tissue water content (TWC) when shoots were exposed to increasing NaCl concentrations (50–200~mM) as compared with the control. Decrease in TWC was not significant at higher NaCl level (150 and 200~mM). At 200~mM NaCl, growth of shoots was adversely affected and microshoots died under prolonged stress. Minimum damage to the membrane as assessed by malondialdehyde (MDA) content was noticed in the controls in contrast to sharp increase of it in NaCl-stressed shoots. Higher amounts of free proline, glycinebetaine and total soluble sugars (TSS) accumulated in NaCl-stressed shoots indicating that it is a glycinebetaine accumulator. About 2.11-fold higher H2O2 content was observed at 50~mM NaCl as compared with control and it reached up to 7.1-folds more at 200~mM NaCl. Antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase and guaiacol peroxidase) also increased with a rise in NaCl level. Increase in bacoside A, a triterpene saponin content was observed only up to 100~mM NaCl level. Higher salt concentrations inhibited the accumulation of bacoside A. It appears from the data that accumulation of osmolytes, ions and elevated activities of antioxidant enzymes play an important role in osmotic adjustment in shoot cultures of Bacopa under salt stress.
  84. Mohan, C. and Kareem, V.K. and Rajasekharan, {\relax PE}, Assessment of Genetic Fidelity in Micropropagated Bacopa Monnieri Using ISSR Marker Assay, Annals of Plant Science, vol. 2, pp. 209--214, January 2013.
    Bacopa monnieri (Linn.) Pennel referred as Brahmi is an important medicinal herb, growing in wet damp and marshy places throughout India. In the present study, nodal explants of B. monnieri were inoculated on MS (Murashige and Skoog) media with or without plant growth regulators (PGRs). In vitro cultures developed were conserved in both Sandard Culture Condition (SCC) and Reduced Culture Conditions (RCC). A large number of plants were regenerated from cultures grown on MS basal medium and MS+PGRs. Inter Simple Sequence Repeat (ISSR) marker assay was employed to validate the genetic fidelity of these regenerated plantlets upto ten passages. Eleven ISSR primers generated a total of 57 amplicons and out of which except one polymorphic band all were monomorphic bands. The dendrogram anlysis (Unweighted Pair-Group Method with Arithmetic averages-UPGMA) revealed that there was no genetic inconsistency exists among the sub-cultured samples on MS basal medium incubated at both SCC and RCC from their mother plant. The results confirmed the clonal fidelity of the tissue culture-raised B. monnieri plantlets and corroborated the fact that multiplication through nodal cuttings using MS basal medium is the safest for multiplication to get true-to-type plants.
  85. Sharma, Munish and Raina, Himani and Vijeshwar, Verma and Mallubhotla, Sharada and Ahuja, Ashok, Synthetic Seeds a Viable Approach for Conservation and Propagation of Phytoremediant Herb: Bacopa Monnieri (l.) Wettst, JERAD, vol. 7, December 2012.
    Bacopa monnieri (L.) Wettst. an emergent macrophyte, fast proliferating wetland species has attracted considerable attention in recent years owing to its capability of absorbing toxic heavy metals from the aquatic environment. As such plant species having dual importance need to be conserved and propagated on priority basis. A protocol was developed for regeneration from encapsulated nodal segments collected from in vitro propagated shoots of Bacopa monnieri. The best gel composition was achieved using 2.5\% sodium alginate and 100 mM CaCl 2 .2H 2 O. The maximum percentage response (86.67) for conservation of encapsulated nodal segments into plantlets was achieved on Murashige and Skoog's medium supplemented with 1.0 mg  -1 BAP after 6-8 weeks. Well developed regenerated plantlets were hardened, acclimatized and established under net house conditions, where they grew well without any detectable variation. The regeneration ability of encapsulated nodal segments was affected by the concentration of sodium alginate and calcium chloride.
  86. Jain, Neelam and Sharma, Varsha and Ramawat, Kishan G., Shoot Culture of Bacopa Monnieri: Standardization of Explant, Vessels and Bioreactor for Growth and Antioxidant Capacity, Physiology and Molecular Biology of Plants, vol. 18, no. 2, pp. 185--190, April 2012. doi: 10.1007/s12298-012-0103-0.
    Standardization of biomass production in different vessels and bioreactor using explants and media for growth, total phenolic content and antioxidant capacity of shoot culture of Bacopa monnieri is described. Maximum number of shoots per explant, higher explants response irrespective of the type of explants, and higher shoot length was obtained on MS medium containing BAP (2.5~mg~l−1) and IAA (0.01~mg~l−1) with 3~\% sucrose. This medium was selected by varying BAP concentration and recorded optimal for shoot culture on gelled medium. The condition of 0.5~cm explant size and 20 explant/40~ml (1 explant/2~ml) was optimal for high explant response, number of shoots per explant regenerated and shoots length. Among the different vessels used, maximum growth index was achieved in Growtek bioreactor (10.0) followed by magenta box (9.16), industrial glass jar (7.7) and conical flask (7.2). The cultures grown in conical flask (100~ml) were used as control. The total phenolic content and antioxidant capacity of in vitro grown plants was higher to that recorded for in vivo material. Among in vitro regenerated plants, the activity was maximal in the tissues grown in 250~ml conical flask. The most critical function for vessels is to support the optimum profusion (growing area for maximum growth) of shoots and for B. monnieri, Growtek bioreactor supported 1980 shoots l−1 medium as compared to control (938 shoots l−1). Growtek bioreactor was considered effective system to produce B. monnieri biomass in culture without loss of antioxidant properties.
  87. Mehta, J. and Kumar, Vinod and Syedy, Mohsina and Upadhyay, Deeksha and Ansari, R. and Bisht, Vaishali and Naaz, Huma and Soni, P., In Vitro Shoot Regeneration of Bacopa Monnieri (L.) Using Cyanobacterial Media- A Novel Approach and Effect of Phytoregulators on in Vitro Micropropagation, undefined, 2012. url: https://www.semanticscholar.org/paper/In-vitro-Shoot-Regeneration-of-Bacopa-monnieri-(L.)-Mehta-Kumar/afe4d673ef3fb9cea2abe3d36621cb2fb999fbed.
    This review highlights the recent development and n ovel achievements made for the multiple shoots rege neration of Bacopa monnieri in Cyanobacterial medium as well as in MS medium. This review highlights the recent development and n ovel achievements made for the multiple shoots rege neration of Bacopa monnieri in Cyanobacterial medium as well as in MS medium. Shootlets were regenerated from noda l explants of stem through auxiliary shoot proliferat ion. In Cyanobacterial culture medium Bacopa monnie ri survived for long period of time and multiple shoot initiati on was observed in 2mg/l Kn. When Bacopa monnieri w as inoculated in MS medium supplemented with different concentrations of PGR’s, maximum no. of shoots wer e observed in 0.5 mg/l BAP + 2.0 mg/l Kn and 0.5 mg/ l Kn+1.0 mg/l BAP.
  88. Sharma, Neelam and Satsangi, Richa and Pandey, Ruchira and Singh, Rakesh and Kaushik, Nutan and Tyagi, Rishi Kumar, In Vitro Conservation of Bacopa Monnieri (L.) Using Mineral Oil, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 111, no. 3, pp. 291--301, December 2012. doi: 10.1007/s11240-012-0194-x.
    Bacopa monnieri (L.) Wettst., a traditional Indian medicinal plant with high commercial potential, is used as a potent nervine tonic. A slow growth protocol was developed for medium-term conservation using mineral oil (MO) overlay. Nodal segments of B.monnieri (two genotypes; IC249250, IC468878) were conserved using MO for 24~months. Single node explants were implanted on MS medium supplemented with 0.2~mg~l−1 BA and were covered with MO. Subculture duration could be significantly enhanced from 6 to 24~months, on the above medium. Normal plants regenerated from conserved cultures were successfully established in soil. On the basis of 20 random amplified polymorphic DNA and 5 inter-simple sequence repeat primers analyses and bacoside A content using HPLC, no significant reproducible variation was observed between the controls and in vitro-conserved plants. The results demonstrate the feasibility of using MO for medium-term conservation of B.monnieri germplasm without any adverse genetical and biochemical effects.
  89. Pharma, International Journal Of and Issn, Bio Sciences and Naik, P. M. and Praveen, N. and Manohar, S. H. and Murthy, H. N., Effect of Mutagens on the in Vitro Adventitious Shoot Growth and Bacoside a Accumulation in Bacopa Monnieri (l.), 2012.
    Mutations were induced in tissue cultured raised Bacopa monnieri (L.) plantlets by treating in vitro derived leaves with gamma rays at (γ – rays) dose rate of 0, 10, 20, 40 and 80 gray (Gy) and 0.5 \% of ethyl methanesulphonate (EMS) at 0, 0.5, 1.0, 1.5, 2.0 and 2.5 hours (h). Xantha mutants were observed with γ- rays at 10, 20 and 40 Gy. All the leaf explants treated with 80 Gy were either necrotic or irregenerable. Both γ and EMS treatment exhibited a reduction in survival rate, number of adventitious shoots, fresh weight and dry weight (DW). The γ- rays doses at 10, 20 and 40 Gy induced mutation in Bacopa monnieri leaf explants and five lines were obtained with increased in the bacoside A content. The lines obtained are D15 (40 Gy) which produced maximum amount of bacoside A (22.567 mg g-1 DW) followed by B11 (10 Gy), C18 (20 Gy), D17 (40 Gy) and B6 (10 Gy) lines which accumulated 21.051, 19.320, 17.163 and 14.133 mg g-1 DW respectively compared with control (13.750 mg g-1 DW). The leaf explants treated with EMS did not show any significant increase with respect to the bacoside A content.
  90. Mehta, J. and Ansari, R. and Syedy, Mohsina and Khan, S. and Sharma, Sushma and Gupta, N. and Rathore, R. and Vaishnav, Komal, An Effective Method for High Frequency Multiple Sho Ots Regeneration and Callus Induction of Bacopa Monnieri (L.) Pennel.: An Important Medicinal Plant, undefined, 2012. url: https://www.semanticscholar.org/paper/An-effective-method-for-high-frequency-Multiple-sho-Mehta-Ansari/39a48fd8dde8be62a3712eae8f515e19431fc053.
    This review highlights the recent development and achievements made for the multiple shoots regeneration and callus induction of Bacopa monnieri. This review highlights the recent development and achievements made for the multiple shoots regeneration and callus induction of Bacopa monnieri. Shootlets were regenerated from nodal explants of stem through auxiliary shoot proliferation. The induction of multiple shoots from nodal segments were highest in MS medium supplemented with 0.5 mg/l BAP + 2.0 mg/l Kn and 0.5 mg/l Kn+1.0 mg/l BAP. For rooting different concentration of IBA were used and highest rooting was recorded on MS medium supplemented with 2.0 mg/l IBA. The rooted Plantlets were hardened initially in culture room conditions and then transferred to misthouse. Leaf petiole explants were used for the purpose of callus induction. Best growth was observed in MS medium supplemented with 0.25 mg/l 2, 4-D+ 0.5 mg/l Kn and 0.25 mg/l 2,4-D+ 0.1mg/l BAP.
  91. Rout, J. and Sahoo, S. and Ray, S. S. and Sethi, B. and Das, R., Standardization of an Efficient Protocol for in Vitro Clonal Propagation of Bacopa Monnieri L. - an Important Medicinal Plant., undefined, 2011. url: https://www.semanticscholar.org/paper/Standardization-of-an-efficient-protocol-for-in-of-Rout-Sahoo/5066a6bb35d86c210996ac84d6478dd10a86bbf2.
    The present study evaluated the most suitable concentration of growth regulators for callus induction and subsequent organogenesis in Bacopa monnieri L. (commonly known as Brahmi) to find the bestcallus induction in MS medium supplemented with 2.0 mg l -1 BAP -1 NAA from leaf explant. The present study was undertaken to evaluate the most suitable concentration of growth regulators for callus induction and subsequent organogenesis in Bacopa monnieri L. (commonly known as Brahmi). The best callus induction (71±2.2\%) was found in MS medium supplemented with 2.0 mg l -1 BAP + 0.5 mg l -1 NAA from leaf explant. Combination of 2.0 mg l -1 BAP + 0.5 mg l -1 NAA and 2.0 mg l -1 BAP + 0.5 mg l -1 IAA gave the most effective for shoot regeneration from callus. The elongated shoots rooted in 1⁄2 strength MS medium supplemented with different concentrations of auxins (NAA, IAA and IBA). IAA (6.5±0.57) was more suitable for root induction when compared to NAA and IBA (5.1±0.32 and 4.7±0.44). The success of plant tissue culture for in vitro culture of B. monnieri was encouraged by acclimatization of the plantlets in the field conditions. About 86\% plantlets survived under field conditions.
  92. Ramesh, Manikandan and Vijayakumar, Karuppa Pillai and Karthikeyan, Alagarsamy and Pandian, Shunmugiah Karutha, RAPD Based Genetic Stability Analysis among Micropropagated, Synthetic Seed Derived and Hardened Plants of Bacopa Monnieri (L.): A Threatened Indian Medicinal Herb, Acta Physiologiae Plantarum, vol. 33, no. 1, pp. 163--171, January 2011. doi: 10.1007/s11738-010-0534-6.
    Bacopa monnieri (L.), a highly endangered miracle medicinal herb with global interest, is one of the popular ancient Indian ayurvedic plants. With ever increasing demand for Bacopa based formulations in pharmaceutical industries, there is a need to preserve the stocks of the plant through biotechnological approaches. Randomly amplified polymorphic DNA (RAPD) fingerprinting approach was applied to analyze the genetic stability of 19 different B.monnieri plants randomly selected after micropropagation, regrowth from alginate encapsulated uninodal cuttings (before and after storage at 4°C) and hardening with the mother plant (wild type). 16 arbitrary decamer primers amplified a total of 334 reproducible distinct DNA fragments ranging from 180 to 1,500~bp, of which 262 (78.4\%) were monomorphic and the rest (21.5\%) were polymorphic with an average of 20.8~bands per primer. The extent of polymorphism was low to moderate. Primers OPAK 14, OPM 15 and OPD 13 generated 69, 46 and 42\% polymorphic patterns. Primers OPA 04, OPU 13 and OPD 08 generated 100\% monomorphic pattern. Similarity matrix based on Jaccard’s coefficient revealed that pair wise values between the wild type and its analyzed plants ranged from 0.00 to 0.92 and among the micropropagated, synthetic seed derived and hardened plants, the range of genetic distance is from 0.67 to 0.92. Unweighted pair group method with arithmetic averages cluster analysis resulted in one loose group of the wild type with three subgroups. The present study paves the way for the identification and maintenance of genetically uniform B. monnieri plants micropropagated in the lab, plants regrown from synthetic seeds and hardened in the field.
  93. Soundararajan, T. and Karranuakaran, C. M., Micropropagation of Bacopa Monnieri through Protoplast, Asian Journal of Biotechnology, vol. 3, no. 2, pp. 135--152, 2011. doi: 10.3923/ajbkr.2011.135.152.
    Micropropagation of Bacopa monnieri through Protoplast
  94. Naik, Poornananda Madhava and Manohar, Shirugumbi Hanamanthagouda and Murthy, Hosakatte Niranjana, Effects of Macro Elements and Nitrogen Source on Biomass Accumulation and Bacoside A Production from Adventitious Shoot Cultures of Bacopa Monnieri (L.), Acta Physiologiae Plantarum, vol. 33, no. 4, pp. 1553--1557, July 2011. doi: 10.1007/s11738-010-0675-7.
    The present work deals with optimization of adventitious shoot culture of Bacopa monnieri for the production of biomass and bacoside A and has investigated the effects of macro elements (NH4NO3, KNO3, CaCl2, MgSO4 and KH2PO4) and nitrogen source [NH4+/NO3−] of Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium (MS) on accumulation of biomass and bacoside A content. Optimum number of adventitious shoots (99.33~shoots~explant−1), fresh weight (1.841~g) and dry weight (0.150~g) were obtained in the medium with 2.0× strength of NH4NO3. The highest production of bacoside A content was also recorded in the medium of 2.0× NH4NO3, which produced 17.935~mg~g−1 DW. The number of adventitious shoot biomass and bacoside A content were optimum when the NO3− concentration was higher than that of NH4+. Maximum number of shoots (70.00~shoots~explant−1), biomass (fresh weight 1.137~g and dry weight 0.080~g) and also bacoside A content (27.106~mg~g−1 DW) were obtained at NH4+/NO3− ratio of 14.38/37.60~mM. Overall, MS medium supplemented with 2.0× NH4NO3 is recommended for most efficient bacoside A production.
  95. Rajeshkumar, T., Effects of Antioxidants and Gelling Agents on Regeneration, in Vitro Conservation and Genetic Stability of Bacopa Monnieri (L.) Pennell., undefined, 2011. url: https://www.semanticscholar.org/paper/Effects-of-antioxidants-and-gelling-agents-on-in-of-Rajeshkumar/21ee0b5ef7afefc77b9fd4864921c988100fdad9.
    These studies provide a feasible in vitro multiplication and conservation method and genetic stability assessment of B. monnieri cultures and no variation in the banding pattern of the cultures grown in different concentrations of growth regulators, agar or isabgol and additives containing media. Shoot regeneration of Bacopa monnieri was achieved, on Murashige and Skoog (MS) medium supplemented with 4.4 μM benzyl aminopurine (BA) from different explants. Nodal explants initiated 75-80 shoots, leaf explants 55-65 shoots and internodal explants 50-55 shoots within 40 days of culture. The initiated shoots were multiplied in MS medium containing 2.2 μM BA. Ascorbic acid (50 mg/l) and citric acid (25 mg/l ) increased the number of shoots and conservation period of the cultures. Isabgol, when used as gelling agent provided consistent multiplication of cultures compared to microbiology grade agar and cultures were conserved in medium containing 3\% (w/v) isabgol for a period of 190 days as against an average of 120 days in 1\% (w/v) agar containing media. The conservation period was increased to a period of 200 days in medium gelled with 3\% (w/v) isabgol containing 50 mg/l ascorbic acid and 25 mg/l citric acid and more than 80\% of the cultures survived in this medium. Rooting was achieved ex vitro by culturing the isolated shoots in autoclaved soilrite moistened with 1/10 strength MS salts. Rooted plantlets were established in soil. Genetic stability assessment with random amplified polymorphic DNA (RAPD) primers showed no variation in the banding pattern of the cultures grown in different concentrations of growth regulators, agar or isabgol and additives containing media. These studies provide a feasible in vitro multiplication and conservation method and genetic stability assessment of B. monnieri cultures.
  96. Mendhulkar, V. and Patade, P. and Singh, S., Dmso Induced Product Recovery of Bacoside a in Cell Suspension Culture of Bacopa Monnieri Linn, 2011. url: https://www.semanticscholar.org/paper/DMSO-INDUCED-PRODUCT-RECOVERY-OF-BACOSIDE-A-IN-CELL-Mendhulkar-Patade/2ea42422d193186951c91e370c52e55b632252d4.
    Bacopa monnieri has been used as anti-depressant, antianxiety, memory enhancer, diuretic, cardio-tonic, in ayurvedic system of medicine, and in modern medicinal formulations. Bacopa monnieri Linn. is an important medicinal plant belonging to family scrophulariaceae. Bacopa is a neurotonic, immuno-modulator, tranquilizing, memory and learning enhancing, cerebral activator, anti-ulcer, antispasmodic, anti-asthmatic. It is also used in Epilepsy, anxiety and depression. Major component of Bacopa are saponins Bacoside A and Bacoside B. The cell suspension culture of Bacopa monnieri was treated with 0.2\%, 0.6\% and 1\% concentrations of DMSO (Dimethyl sulphoxide), a cell permeabilizing agent for 3 and 6 hours. The samples were analyzed for the content of bacoside A using HPTLC technique to identify and quantify the amount of released product. Maximum release of bioactive compound, Bacoside A was observed (4.6±0.02μg/mg) in suspension culture treated with 1\% DMSO for 3 and 6 hrs treatment durations compared to control (2.1±0.05 and 3.2±0.01, respectively). INTRODUCTION: Natural products have been used in the treatment of various human ailments. The most important and oldest treaties of Indian medicine system (i.e., Ayurveda) are Charak Samhita (1000 B.C) and Sushruta Samhita that describes the use of more than 500-700 medicinal herbs. Today plants have contributed more than 7000 different compounds which are useful as heart drugs, laxatives, anti-cancer agents, hormones, anti-parasitic compounds etc. Worldwide extensive screening programs are being carried out to identify chemical compounds from plants that may provide new treatments for various human diseases. In recent years a lot of attention has been devoted to novel molecules derived from natural sources that exhibit a good range of clinical and pharmacological activities. A plant system, Bacopa monnieri Linn. is used in ethnomedicinal and modern medicinal formulations. This plant is commonly known as Brahmi. Bacopa monnieri has been used as anti-depressant, antianxiety, memory enhancer, diuretic, cardio-tonic, in ayurvedic system of medicine. Leaf extracts exhibits the analgesic, anti-inflammatory and anti-pyretic activities . The plant contains two saponins, bacoside A and B. In addition to the bacosides, Bacopa contains a wide variety of medically active substances, including stigmasterol, sapogenins, and flavonoids . Other compounds include triterpenoid saponins. Bacopa also contains D-mannitol, betulic acid, beta-sitosterol, octacosane, nicotine, and amino acids such as alphaalanine, aspartic acid, glutamic acid, and serine. The qualitative and quantitative improvement of biologically active compounds by in-vitro techniques has been achieved most successfully and promisingly in many medicinal plants. Large-scale production of active component from the cell culture, enhancement in the production of secondary metabolites by using chemical and biological elicitors and increasing
  97. Sharma, N. and Satsangi, R. and Pandey, R., Cryopreservation of Shoot Tips of Bacopa Monnieri (l.) Wettst. by Vitrification Technique, Acta Horticulturae, no. 908, pp. 283--288, September 2011. doi: 10.17660/ActaHortic.2011.908.37.
    A protocol for cryopreservation of in vitro shoot tips of Bacopa monnieri (L.) Wettst., a medicinal plant of high commercial value was developed. Shoot tips (about 1 mm in length) excised from 4- to 24-week-old proliferating shoot cultures were precultured on MS medium supplemented with various cryoprotectants before dehydration with PVS2 at 0°C. The dehydrated shoot tips were directly immersed in liquid N2. Following cryopreservation, and after rapid rewarming at 45°C, shoot tips were quickly washed with 1.2 M sucrose solution and then plated on solidified shoot culture medium. Shoot tips were successfully cryopreserved by vitrification, when they were precultured on medium supplemented with 5\% DMSO for two days before dehydration. Preculture of shoot tips with sucrose at 25°C significantly improved the survival and shoot regeneration rate of cryopreserved shoot tips. Regenerated plantlets were successfully transferred to pots.
  98. Banerjee, Meenakshi and Modi, Priyanka, Micropropagation of Bacopa Monnieri Using Cyanobacterial Liquid Medium, Plant Tissue Culture and Biotechnology, vol. 20, no. 2, pp. 225--231, 2010. doi: 10.3329/ptcb.v20i2.6917.
    Hot extract of Aulosira fertilissima (cyanobacterium) added in different proportions to MS as a liquid culture media for the in vitro propagation of Bacopa monnieri (L.) Pennell. Maximum numbers of shoots were induced from axillary node in MS media (40 ml) + Aulosira extract (60 ml) and maximum shoot multiplication was observed when Kn (1.0 mg/l) was added in the shoot initiation media (mentioned above). Surprisingly rooting was also found to be best in the same combination of MS + cyanobacterial extract that was used for initiation and multiplication of shoots. On an average within a period of three subcultures (2 - 3 months) the nodal explants generated 400 shoots. Rooted plantlets were successfully transferred to the field, after acclimation in the net house. Key words: Baccopa monnieri, Cyanobacterial extract, Regeneration, Acclimation D.O.I. 10.3329/ptcb.v20i2.6917 Plant Tissue Cult. \& Biotech. 20(2): 225-231, 2010 (December)
  99. Showkat, Patni and Zaidi, Yaseer and Asghar, Suhail and Jamaluddin, Shamsuddin, In Vitro Propagation and Callus Formation of Bacopa Monnieri (L.) Penn., Plant Tissue Culture and Biotechnology, vol. 20, no. 2, pp. 119--125, 2010. doi: 10.3329/ptcb.v20i2.6890.
    An important medicinal herb, namely Bacopa monnieri (L.) Penn. showed best results on MS with 1 mg/l IAA and 1 mg/l IBA for culture initiation, multiple shoot proliferation, bud breakage, shoot length, shoot numbers and cluster formation. For rooting the best result was obtained on MS solidified with 7 g/l of agar along with sugar 20 gm/l for root, shoot ratio and number of roots. The best hardening was achieved in a mixture of soil : soilrite (1 : 1) where the survival rate was 94\%. Further, for callus formation the best result was obtained in the leaf explants on MS supplemented with 0.5 mg/l 2, 4-D. HPLC and HPTLC profile of regenerated shoots revealed a phytochemicals profile similar to that of the market samples and mother plants. The presence of bacoside was detected through HPTLC and HPLC. Key words: Bacopa monnieri, Micropropagation, Bacoside D.O.I. 10.3329/ptcb.v20i2.6890 Plant Tissue Cult. \& Biotech. 20(2): 119-125, 2010 (December)
  100. Antony Ceasar, S. and Lenin Maxwell, S. and Bhargav Prasad, K. and Karthigan, M. and Ignacimuthu, Savarimuthu, Highly Efficient Shoot Regeneration of Bacopa Monnieri (L.) Using a Two-Stage Culture Procedure and Assessment of Genetic Integrity of Micropropagated Plants by RAPD, Acta Physiologiae Plantarum, vol. 32, no. 3, pp. 443--452, May 2010. doi: 10.1007/s11738-009-0419-8.
    A two-stage culture procedure has been developed for highly efficient shoot regeneration from leaf and internode explants of Bacopa monnieri. Adventitious shoot buds were obtained on the shoot induction medium containing Murashige and Skoog’s (MS) basal salt supplemented with 1.5~mg/l thidiazuron and 0.5~mg/l naphthalene acetic acid; these shoot buds were subcultured on the multiplication (second) medium amended with BAP (benzyl amino purine). Multiplication medium containing 0.5~mg/l BAP produced more shoots (135) and longer shoots (7.8~cm) with more nodes (6). Best response of root induction with more number of roots (16.5) and longer roots (8.7~cm) was observed in half strength MS basal medium supplemented with 1.0~mg/l IBA (indole-3-butyric acid) and 0.5~mg/l phloroglucinol. In vitro obtained plants were transferred to the field after hardening with a 100\% survival rate. Random amplified polymorphic DNA analysis was carried out using five random primers. The amplification products were monomorphic in micropropagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic integrity of micropropagated plants.
  101. Naik, Poornananda Madhava and Manohar, Shirugumbi Hanamanthagouda and Praveen, Nagella and Murthy, Hosakatte Niranjana, Effects of Sucrose and pH Levels on in Vitro Shoot Regeneration from Leaf Explants of Bacopa Monnieri and Accumulation of Bacoside A in Regenerated Shoots, Plant Cell, Tissue and Organ Culture (PCTOC), vol. 100, no. 2, pp. 235--239, February 2010. doi: 10.1007/s11240-009-9639-2.
    Brahmi (Bacopa monnieri) is an important medicinal plant mainly used for the treatment of neurological disorders and depression. Recent investigations revealed that bacoside A is major chemical component shown to be responsible for memory facilitating action of brahmi. The current investigation was carried out to assess the potential for increasing biomass and the concentration of bacoside A in the in vitro regenerated shoots by varying sucrose and pH levels of shoot regeneration medium. The leaf explants were cultured on the Murashige and Skoog (MS) medium supplemented with 2~mg~l−1 kinetin (KN) and with varying concentrations of sucrose (0, 1, 2, 3, 4, 5 and 6\% at pH 5.8) and pH (4.5, 5.0, 5.5, 6.0 and 6.5 with 2\% sucrose) with the objective of verifying the effects of sucrose and pH level on shoot regeneration and to verify the accumulation of bacoside A in the regenerated shoots. The shoot biomass increased (150.50~±~2.84 shoots per explant, fresh wt 6.31~±~0.12~g and dry wt 250~±~5.00~mg) on the medium supplemented with 2\% sucrose and pH which was set at 4.5. The results of HPLC analysis indicate that increase in sucrose concentration (0, 1, 2, 3, 4, 5 and 6\% at pH 5.8) lead to decrease in the bacoside A content (39.51, 22.43, 13.05, 12.17, 10.73, 9.56 and 8.93~mg~g−1 dry wt, respectively) in regenerated shoots. These findings provide evidence that stressful condition of inadequate supply of carbon elevated synthesis of bacoside A in brahmi shoots. However, 2\% sucrose is found suitable for biomass accumulation. Therefore, medium supplemented with 2\% sucrose and pH set at 4.5 was found suitable for both biomass (6.31~±~0.12~g fresh wt and 250~±~5.00~mg dry wt) and bacoside A accumulation (13.09~mg~g−1 dry wt).
  102. Soundararajan, T., Micropropagation Studies on Bacopa Monnieri, Nerium Oleander and Bioreactor Studies on Micropropagation of Dendrobium, an Orchid, April 2009. url: http://hdl.handle.net/10603/51.
    Orchids are an excellent item for garden and for indoor decoration. They exhibit a wide range of diversity in form, size, color and texture of flowers beyond the imagination of human mind (Mukherjee, 1983). Orchids have been cultivated over past 200 years for their beautiful flowers. They belong to one of the largest families of plants on earth. Several orchids have been used as food in different parts of the world (Bose and Bhattacharjee,1999). Orchids grow in nature through seeds but in absence of appropriate hosts they don t germinate in adequate numbers, so it remains as a rare species For appropriate germination of orchid seeds, in vitro propagation is imperative for multiplication rather than in vivo (Arditti,1979). Clonal propagation of plants is often difficult, expensive and unsuccessful. Clonal propagation through tissue culture popularly called Micropropagation . A number of hybrids of commercial importance are routinely propagated by tissue culture methods. Use of plant tissue culture for micropropagation was initiated and he found commercially viable approach for orchid propagation. Commercial in vitro propagation of plants is done essentially by agar based media. Though this procedure has advantages over conventional means of muliplicaton in terms of cost and labour. This aspect obviously lead to vigorous search for less labour and cost effective process. Use of bioreactors wherein cultures can be maintained in precisely controlled conditions with higher culture volumes has been the natural choice for adaptation since they have been utilized for culturing plant cells and aggregates for the production of secondary metabolites. Physical and chemical culture conditions are crucial in large scale micropropagation of clonally propagated plant species. ii The use of bioreactors for plant multiplication is a novel and developing technology. Realization of the potential of bioreactors for the plant multiplications have led to demonstrations with an increasing number of plant systems.
  103. Prakash, Parale Anuradha and Dayaram, Nikam Tukaram, Influence of Auxins, Cytokinins and Biotic Elicitors on Accumulation of Memory Enhancer Compound Bacoside-A in Tissue Culture of Bacopa Monniera (L.) Pennell., Medicinal and Aromatic Plant Science and Biotechnology, vol. 3, no. 1, pp. 74--81, 2009.
    The effects of the auxins [indole-3-acetic acid (IAA), 1-napthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D)], cytokinins [6-benzyladenine (BA), Kinetin (Kin) and thidiazuron (TDZ)] and biotic elicitors were evaluated individually and in combinations, on callus proliferation, shoot growth and accumulation of bacoside-A through liquid and semi-solid nutrient media. Maximum shoot proliferation (19.4 ± 0.91 shoots/explant) was observed in liquid MS medium containing 5 μM BA, which was 1.3 times higher than semi-solid MS medium fortified with same concentration of BA. The callus and shoots showed sustained growth and accumulation of bacoside-A even after two years of initiation of subcultures. The nature of nutrient media did not influence the accumulation of bacosideA in shoot biomass. Except for Saccharomyces cerevisiae-derived elicitors that showed about a 20\% increase in shoot biomass production, the growth of callus as well as shoots was inhibited after the addition of other fungal elicitors in MS medium. Among the shoot and callus cultures exposed to Pencillium notatum, Rhizopus stolonifer, Coriolus versicolor, Mucor sp. and S. cerevisiae-derived elicitors, incorporation of 750 mg/l S. cerevisiae or Mucor sp. showed about 3.2- and 1.7-fold higher production of bacoside-A in callus and shoot biomass, respectively. This protocol would be helpful in reducing pressure on natural populations of Bacopa monniera (L.) Pennell. harvested indiscriminately for bacoside-A.
  104. Ramseh, M and Marx, R and Mathan, G and Pandian, S Karutha, Effect of Bavistin on in Vitro Plant Conversion from Encapsulated Uninodal Microcuttings of Micropropagated Bacopa Monnieri (L.) – An Ayurvedic Herb, pp. 4, 2009.
    A reliable and reproducible protocol for contamination free plant recovery system from alginated encapsulated uninodal microcuttings of micropropagated Bacopa monnieri L.have been developed after storage at 18oC for 45 days. Node segments excised from freshly micropropagated plants were encapsulated as single explant beads with 3.0 \% sodium alginate and 80 mM CaCl 2 H O. To find out the 2 2 optimal concentration of fungicide bavistin for efficient plant recovery, different concentrations of bavistin (1.0 - 15 mg l-1) were incorporated in to the encapsulation medium. 3.0 mg l-1 bavistin showed no reduction in plant conversion and generated maximum number of shoots (45.6 ± 1.69) at high frequency with out any contamination after storage up to 45 days at 18oC. At high concentrations (13 and 15 mg l-1), rupturing of calcium alginate coats after 8 - 9 days and gradual decline in the number of shoots indicates the toxic effect of bavistin on plant conversion. Encapsulated node cuttings stored up to 45 days regenerated shoots (5.2) and multiple shoots (45.6) in MS basal and hormone medium respectively. Maximum shoot length (8.2 ± 0.37 cm) was observed from encapsulsted node cuttings incorporated with 3.0 mg l-1 bavistin on MS basal medium. 90 \% of the recovered plantlets were hardened off and successfully established in soil.
  105. Banerjee, Meenakshi and Shrivastava, Sarika, An Improved Protocol for in~Vitro Multiplication of Bacopa Monnieri (L.), World Journal of Microbiology and Biotechnology, vol. 24, no. 8, pp. 1355--1359, August 2008. doi: 10.1007/s11274-007-9612-3.
    A cost-effective and efficient protocol has been described in the present work for large scale and rapid in~vitro propagation of a valuable medicinal herb Bacopa monnieri (L.) by shoot bud proliferation on internodal segments isolated from field grown mature plants. This shooting was achieved on low concentration of cytokinin with BAP 1.0~mg/l and Kn 0.5~mg/l within 3~weeks of inoculation contrary to earlier reports where auxin is also used in combination with cytokinin. Normally internodes gives rise to a clump of shoots (18 shoots). On an average within a period of three subcultures the internode explant generated 324 shoots thereby improving the economics of the cost of the plants produced and time required. For rooting this is a first report of its growth in a liquid culture. The regenerated plants resemble the mother plants in the general habitat without any morphological variation. This paper reports a very simple, reproducible procedure for in~vitro propagation of Bacopa monnieri (L.) that can be used to form a foundation stock of elite plant material for large-scale cultivation.
  106. Sharma, Neelam and Satsangi, Richa and Pandey, Ruchira and Vimala Devi, S., In Vitro Clonal Propagation and Medium Term Conservation of Brahmi [Bacopa Monnieri (L) Wettst], Journal of Plant Biochemistry and Biotechnology, vol. 16, no. 2, pp. 139--143, July 2007. doi: 10.1007/BF03321990.
    A protocol has been developed for in vitro clonal propagation leading to conservation of Bacopa monnieri (L) Wettst, a medicinal plant of high commercial potential with legendary reputation as a memory vitalizer. Single node explants when cultured on Murashige and Skoog’s medium supplemented with BA (0.2 mg l-1), showed active shoot proliferation (22.2 shoots/ explant in 8 weeks) without callus formation. Rooting was achieved on the same medium. The generated shoots could also be conserved for 12 months with high survival rate (up to 100\%). The regenerants upon transfer to soil showed no morphological variation as compared with the donor plants. The medium optimized in the present study was applied for culture establishment and conservation of a total of 15 Brahmi accessions procured from different regions.
  107. Salvio Escandón, Alejandro and Hagiwara, Juan Carlos and Alderete, Liliana Marisol, A New Variety of Bacopa Monnieri Obtained by in Vitro Polyploidization, Electronic Journal of Biotechnology, vol. 9, no. 3, pp. 0--0, June 2006. url: http://www.scielo.cl/scielo.php?script=sci_abstract&pid=S0717-34582006000300002&lng=es&nrm=iso&tlng=en.
  108. Mohapatra, H. P. and Rath, S. P., In Vitro Studies of Bacopa Monnieri--an Important Medicinal Plant with Reference to Its Biochemical Variations, Indian Journal of Experimental Biology, vol. 43, no. 4, pp. 373--376, April 2005.
    Micropropagation of Bacopa monnieri was achieved on MS and B5 medium supplemented with BAP and NAA using leaf explants and nodal segments. Best results were found on MS medium in both the explants with BAP (2.0 mg/l) showing higher percentage of regeneration. Besides that the biochemical parameters, like chlorophyll, carbohydrate, protein, of leaves both in vivo and in vitro have also been carried out in order to establish the sustainability of plants.
  109. Shrivastava, N. and Rajani, M., Multiple Shoot Regeneration and Tissue Culture Studies on Bacopa Monnieri (L.) Pennell, Plant Cell Reports, vol. 18, no. 11, pp. 919--923, August 1999. doi: 10.1007/s002990050684.
    Adventitious shoot buds were induced from leaf and stem explants of Bacopa monnieri on Murashige and Skoog medium supplemented with benzyladenine or kinetin. The source of the explants as well as different gelling agents in the medium were found to influence shoot induction and eventual shoot growth. The best response was obtained in leaf explants taken from shoot cultures grown in medium supplemented with 2\,μM benzyladenine and gelled with 0.2\% gelrite. A transverse section of the leaf explant incubated in this medium showed several shoot primordia emerging from the leaf surface. This system exhibited a potential for repeated harvesting of the shoots from the original leaf explant as the latter continued to expand and regenerate new shoots, upon repeated periodical subculturing onto fresh medium. However, the callusing response of the plant was very low. Qualitative TLC studies of the regenerated shoots revealed a phytochemical profile similar to that of the field grown-plants.
  110. Sharma, Munish and Koul, Anuja and Ahuja, Ashok and Mallubhotla, Sharada, Suitability of Bench Scale Bioreactor System for Shoot Biomass Production and Bacoside Biosynthesis from Bacopa Monnieri (L.), Engineering in Life Sciences, vol. 19, pp. 584--590, 2019. doi: 10.1002/elsc.201800184.
    According to folklore, Bacopa monnieri commonly called as Brahmi is known for its cognitive enhancing properties. The plant is found abundantly in wetlands but the drug content (bacosides) is very low (0.2\%), therefore, alternative biotechnological protocols are highly needed to supplement the constant source of this valuable plant material which produces stable amounts of bacosides. The present study was conducted to explore the application of different culture systems for cultivation of shoot biomass and maximization of biologically active bacoside biosynthesis in this medicinally important plant. Shoot cultures of Bacopa were cultivated in two different modified benchtop bioreactors: glass bottle bioreactor and balloon type bubble bioreactor and compared with those grown in traditional Erlenmeyer agitated flask. The shoots cultivated in the balloon type bubble bioreactor system showed excellent growth (growth index 796.47 ± 17.27 fresh weight and 395.55 ± 7.55 dry weight) as compared to glass bottle bioreactor system (growth index 488.17 ± 14.4 fresh weight and 327.79 ± 6.64 dry weight) and agitated flask (growth index 363.43 ± 11 fresh weight and 304.22 ± 6.76 dry weight). Furthermore, bacosides produced by shoot cultures cultivated in the balloon type bubble bioreactor (321.95 ± 17.14 mg/L) and glass bottle bioreactor (180.18 ± 6.25 mg/L) configurations were ∼2.78 fold and ∼1.55 fold higher than that recorded in agitated flask cultures (115.7 ± 3.84 mg/L). The balloon type bubble bioreactor system was found to be advantageous for enhancing B. monnieri shoot biomass and bacoside biosynthesis along with ensuring a successful protocol for continuous supply.
  111. Kalita, Bipul Ch and Doley, Pradip and Mitra, P K and Tag, Hui and Das, A K, Optimization of Ms Media for Micropropagation of an Medicinal Plant-Bacopa Monnieri (l.), World Journal of Pharmacy and Pharmaceutical Sciences, vol. 7, no. 6, pp. 1014--1022, 2018. url: https://storage.googleapis.com/journal-uploads/wjpps/article_issue/1527761731.pdf.
    Bacopa Monnieri L. popularly called as Brahmi is an important medicinal plant, traditionally used as tonic to enhance memory development, learning, concentration and to provide relief to patients with anxiety or epileptic disorders (Debnath et al, 2006).
  112. Haque, Sk Moquammel and Ghosh, Biswajit, Micropropagation, in Vitro Flowering and Cytological Studies of Bacopa Chamaedryoides, an Ethno-Medicinal Plant, Environmental and Experimental Biology, vol. 11, pp. 59--68, 2013. url: https://www.researchgate.net/publication/265601937_Micropropagation_in_vitro_flowering_and_cytological_studies_of_Bacopa_chamaedryoides_an_ethno-medicinal_plant.
    This is the first report of both establishment of micropropagation protocol as well as cytological studies of Bacopa chamaedryoides (Kunth) Wettst., an important Indian ethno-medicinal herb. Shoot-tips and nodal segments explants were inoculated on Murashige and Skoog basal medium containing 0.7\% agar, 3.0\% sucrose and different concentrations and combinations of cytokinins and auxins. Optimum multiplication was achieved on medium containing 6-benzyl-aminopurine (2.0 mg L–1) and indole-3-acetic acid (0.2 mg L–1). Shoot-tips proved to be a better explant in having a high rate of shoot multiplication (18.7 ± 0.17) in comparison to nodal segments (15.1 ± 0.18) in the same medium. In vitro rooting of multiplied individual shoots was achieved on half strength Murashige and Skoog medium supplemented with 50\% of ‘Aloe vera gel’, with a maximum of 18.3 ± 0.17 roots. Up to 66.7\% of these multiplied shoots induced healthy flowers in vitro on Murashige and Skoog medium containing low concentration of 6-benzyl-aminopurine (0.2 mg L–1). In vitro produced flowers contained 96.54 \% viable pollen, more or less same as the field grown mother plants. Micropropagated plants have shown normal diploid 2n = 22 chromosomes, same as that of the mother plant. These micropropagated plants were successfully established in soil after hardening them in submerged condition, with an 84\% survival rate. In total, 88.9\% of the survived plants flowered and fruited normally after 50 days of field transfer. More than 85\% field-grown regenerated plants developed normal fruits and viable seeds. This work presents an efficient micropropagation method from node and shoot-tip culture for mass propagation and in vitro flowering of B. chamaedryoides.
  113. Varghese, Dalia B and Sathyanarayana, B N, Induction of Variations in Two Cultivars of Bacopa Monnieri by Gamma Irradiation of In Vitro Cultures, Medicinal and Aromatic Plant Science and Biotechnology, vol. 1, no. 1, pp. 86--89, 2007. url: http://www.globalsciencebooks.info/Online/GSBOnline/images/0706/MAPSB_1(1)/MAPSB_1(1)86-89o.pdf.
    In vitro cultures of nodal segments and calli of two cultivars of Bacopa monnieri viz., ‘Pragyashakthi’ and ‘Calcutta Local’ were subjected to gamma irradiation (30 Gy to 100 Gy for nodal segments and 30 Gy to 80 Gy for calli) with a view to induce variations and to assess the sensitivity of the cultivars to the mutagen treatment. The mutagenic treatments generated substantial variability in the morphological characters of the regenerated plantlets. Variation in leaf morphology induced by the mutagen was observed in plantlets of both the cultivars, while distinctly dwarf plantlets were observed only in case of ‘Calcutta Local’ with a high gamma ray dose of 100 Gy. The results indicate that both cultivars respond differently towards mutagenic treatments, visible from the frequency of mutations induced. Nodal segments of ‘Calcutta Local’ showed a higher mutation frequency while, in the case of calli, ‘Pragyashakthi’ was more sensitive to gamma irradiation. Such variation between cultivars depending on the type of explant used has previously never been reported. Variation was also observed with regard to the bacoside (active principle) content of the plantlets derived from the gamma-irradiated explants. Variants with high content of bacoside (to a maximum of 3.03\%) could be isolated from the fourth subculture in the case of both the cultivars. The encouraging results obtained from the present investigation emphasize the efficiency of in vitro mutation induction for the improvement of this important medicinal plant.
  114. Bansal, Y. K. and Kushwaha, A. K., In Vitro Regeneration of ‘Brahmi ’[Bacopa Monnieri (L.) Pennel] and Preliminary Analysis of Its Secondary Metabolites-Indian Journals, Journal of the Indian Botanical Society, vol. 92, no. 1\&2, pp. 15--20, 2013. url: https://www.indianjournals.com/ijor.aspx?target=ijor:jibs&volume=92&issue=1and2&article=003.
    A standard protocol for in vitro regeneration of Bacopa monnieri has been successfully developed. Best shoot initiation and multiple shooting were obtained on BAP 5mg L−1 from leaf explants, whereas liquid basal medium produced maximum number of roots. In vitro regenerated plants were successfully hardened and about 90\% plants survived on field transfer. The saponin content in both the nature grown and in vitro raised plants were analyzed. In vitro raised plants were found to have low saponin content.
  115. Sharma, Sudhir and Kamal, Barkha and Rathi, Neelima and Chauhan, Sudhir and Jadon, Vikas and Gehlot, Ashok and Arya, Sarita, In Vitro Rapid and Mass Multiplication of Highly Valuable Medicinal Plant Bacopa Monnieri (L.) Wettst., African Journal of Biotechnology, vol. 9, no. 49, pp. 8318--8322, December 2010. doi: 10.4314/AJB.V9I49.
    A protocol has been developed for micropropagation of Bacopa monnieri (L) Wettst, a medicinal plant of high commercial potential with legendary reputation as a memory vitalizer. Nodal segments containing axillary buds were surface sterilized with 0.1\% solution of mercuric chloride for 5 min and were inoculated aseptically on culture medium, axillary bud break was achieved in 100\% of cultures in semisolid MS medium supplemented with 0.2 mg/l BAP. These proliferated in vitro axillary shoots were excised and cut into groups of shoot clusters and subcultured on MS medium supplemented with 0.2 mg/l BAP for shoot multiplication. 100\% in vitro rooting was obtained when shoot clusters were cultured on MS medium supplemented with 0.15 mg/l IBA. The rooted plantlets were hardened, acclimatized and successfully established in field.
  116. Nagarajan, T and Alagumanian, S and Jahirhussain, G and Subbaiya, S, In Vitro Mass Propagation of Bacopa Monnieri (Linn.) Wettst from Nodal Explant - a Multipurpose Medicinal Plant, World Journal of Pharmaceutical Research, vol. 4, no. 12, pp. 1970--1982, 2015.
    Bacopa monnieri is a highly valuable medicinal plant from the Scorphularaceae family. It is used to derivate different pharmaceutical products, but it lost from their natural habitats due to indiscriminate collection for pay pharmaceutical demands. Therefore, the present study was carried out to determine the in vitro mass propagation of Bacopa monnieri (Linn.) Wettst. by using nodal explants. Nodal explants of Bacopa monnieri were cultured on MS basal medium supplemented with different concentrations of BAP and KIN ranging from 0.5-2.5 μM/L for multiple shoot induction. Two cytokinins tested, BAP was found to develop in shoot multiplication and higher number of shoots from the nodal explants when compared to KIN. Higher number of shoot was produced from all the concentrations of both BAP and KIN. The highest frequency (100\%) of shoot induction and maximum number of shoot (6.4±1.94) was observed on 2.0 μM BAP with shoot length of 5.02±0.20 c.m. as well as in KIN the highest frequency (100\%) of shoot induction and maximum number of shoot (5.4±1.51) was observed on 1.5 μM KIN with shoot length of 5.14±0.28 c.m. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of IBA and NAA for root induction. The rooted plantlets were successfully transferred in soil through hardening and established in the field.
  117. Umesh, T.G. and Sharma, Ashwani and Rao, N.N., Regeneration Potential and Major Metabolite Analysis in Nootropic Plant-Bacopa Monnieri (L.) Pennell, Asian Journal of Pharmaceutical and Clinical Research, vol. 7, pp. 134--136, January 2014.
    OBJECTIVE: Bacopa monnieri (L) Pennell, commonly known in India as 'Brahmi' is an important nootropic plant. The study was mainly aimed at the mass multiplication of B. monneri and the effect of various growth regulator regimes for their active principle accumulation in regenerated plants. METHOD: In this study, a protocol to produce high frequency of in vitro cultured multiple shoots / plantlets were established both by direct and callus based indirect regeneration using leaf and nodal explants. In direct regeneration, a single leaf grown on MS basal plant medium supplemented with 2mg/L kinetin gave large number of multiple plantlets. On the other hand the callus based regeneration using leaf as explant grown on MS basal plant medium supplemented with 1mg/L Benzyl aminopurine (BAP) and 0.5mg/L IAA produced large number of side shoots / multiple plantlets. The successfully acclimatized plants were screened for major metabolites /active principles using HPLC. RESULT: Influence of Cytokinins and supplementation with auxins for the production of multiple shoots was investigated. 2mg/L kinetin alone showed highest number of shoots whereas the combination of Kin / BAP or Kin / IAA showed less number of shoots. Bacopaside content of regenerated shoots varied with hormonal treatments. CONCLUSION: This study will facilitate the mass propagation and transformation of this nootropic plant. In addition the study can be used for commercial production of secondary metabolites like Bacopaside using different hormone combinations for elicitation.
  118. Kunakhonnuruk, Boworn and Inthima, Phithak and Kongbangkerd, Anupan, Improving Bacoside Yield of Bacopa Monnieri (L.) Wettst. in Temporary Immersion System by Increasing Immersion Time and Lowering the Intervals, Industrial Crops and Products, vol. 191, pp. 115859, January 2023. doi: 10.1016/j.indcrop.2022.115859.
    Bacopa monnieri (L.) Wettst. or Brahmi has been used in Ayurvedic medicine and currently use as an alternative medicinal herb for several therapeutic healing. Therefore, in vitro mass propagation of B. monnieri for high quality plant production using temporary immersion system (TIS) was necessarily examined. Explants of B. monnieri as regenerated shoots from leaf segments were used to determine the effects of culture system, immersion time and inoculum explant density on growth and bacoside production of B. monnieri. Statistically significant differences in growth and biomass accumulation were observed in the TIS compared with continuous immersion and semi-solid systems. Production of bacoside A, bacopaside II, bacopasaponin C, and total bacoside in B. monnieri obtained from microcontainer cultivation was significantly higher than TIS, whereas total yield of bacosides obtained from different culture systems showed no significant differences. Three immersions per day for 10~min duration promoted growth and biomass accumulation of B. monnieri, while cultivation with 20 explants per container offered optimal inoculum density for large-scale production. Various concentrations and yields of bacoside were obtained from different immersion interval conditions. Reducing immersion duration improved bacoside production. Five explants per container provided the highest bacoside content but yield per container was the lowest.
  119. Soni, Sumit K. and Singh, Rakshapal and Ngpoore, Nem K. and Niranjan, Abhishek and Singh, Purnima and Mishra, Aradhana and Tiwari, Sudeep, Isolation and Characterization of Endophytic Fungi Having Plant Growth Promotion Traits That Biosynthesizes Bacosides and Withanolides under in Vitro Conditions, Brazilian Journal of Microbiology, vol. 52, no. 4, pp. 1791--1805, December 2021. doi: 10.1007/s42770-021-00586-0.
    Endophytes are regarded with immense potentials in terms of plant growth promoting (PGP) elicitors and mimicking secondary metabolites of medicinal importance. Here in the present study, we explored Bacopa monnieri plants to isolate, identify fungal endophytes with PGP elicitation potentials, and investigate secretion of secondary metabolites such as bacoside and withanolide content under in vitro conditions. Three fungal endophytes isolated (out of 40 saponin producing isolates) from leaves of B. monnieri were examined for in vitro biosynthesis of bacosides. On morphological, biochemical, and molecular identification (ITS gene sequencing), the isolated strains SUBL33, SUBL51, and SUBL206 were identified as Nigrospora oryzae (MH071153), Alternaria alternata (MH071155), and Aspergillus terreus (MH071154) respectively. Among these strains, SUBL33 produced highest quantity of Bacoside A3 (4093~μg~mL−1), Jujubogenin isomer of Bacopasaponin C (65,339~μg~mL−1), and Bacopasaponin C (1325~μg~mL−1) while Bacopaside II (13,030~μg~mL−1)~was produced by SUBL51 maximally. Moreover, these aforementioned strains also produced detectable concentration of withanolides—Withaferrin A, Withanolide A (480~μg~mL−1), and Withanolide B (1024~μg~mL−1) respectively. However, Withanolide A was not detected in the secondary metabolites of strain SUBL51. To best of our knowledge, the present study is first reports of Nigrospora oryzae as an endophyte in B. monnieri with potentials of biosynthesis of economically important phytomolecules under in vitro conditions.
  120. Zaidi, Tasawwur Husain, Cultivation of Brahmi (Bacopa Monniery Linn.) in Haryana: An Agroecological Approach, pp. 13, 2018.
    Brahmi (Bacopa monniery Linn.) is being cultivated in many states of north India but there is a gap between demand and supply. This gap can be filled by bringing more and more suitable areas under its cultivation by studying its agroecological requirements. This plant is used in many medicines and health products and its market demand is also growing consistently. It provides opportunity for further expansion of the cultivable land under this plant. Wetlands, ponds and damp areas may also be suitably used for the cultivation of this plant. Irrigation facilities in the state are also well developed. These factors in combination with agroecological suitability of cultivating this plant in the state of Haryana have prompted this investigation. In the present research agroecological approach has been adopted for scientific investigation of the areas as well as agricultural condition which support the cultivation of this plant. The basic assumption underlying this approach is that every region with its unique natural and geographic conditions has potential to supports different species and varieties of crops and plants. These potentials should be properly understood and utilised for the benefit of society and natural environment. In the present paper, agroecological conditions (climatic, pedological and agricultural) of cultivating brahmi in Haryana have been studied. For this purpose agroecological requirements or sets of potentials and constraints (land, soil and environmental characteristics) for the cultivations of brahmi have been used to identify zones/regions of varying suitability in GIS environment using IDW interpolation method of spatial analysis kit in ArcGIS 10.3 software. This exercise has yielded four zones of different levels of suitability on the basis of average expected yield of the plants in different locations of the state. Around 15 per cent area of the state has been found to be more than just suitable for the cultivation of this plant. Around 54 per cent area in the western part of the state is not suitable for this plant. Wetlands, ponds, lakes, river banks, canal sides form special areas for the cultivation of this plant.
  121. Varinder, Singh and Naman, Jain and Richa, Shri, Effect of Abiotic Factors on Bacoside A Content, Acetylcholinesterase Inhibitory and Antioxidant Activities of Bacopa Monnieri (L.) Wettst, Journal of Applied Pharmaceutical Science, March 2021. doi: 10.7324/JAPS.2021.11s107.
    Growth of plants and production of phytoconstituents are influenced by abiotic stresses. Understanding these abiotic factors and their subsequent modification during cultivation/growth of medicinal plants may help in increasing the production of valuable secondary metabolites. The present study examined the effect of various abiotic factors and seasons on the growth of Bacopa monnieri, production of the marker compound bacoside A, and antioxidant and acetylcholinestrase inhibitory activities of the plant extract. The plant was cultivated in two different soils, viz A (clay loam soil) and B (sandy loam soil). Different abiotic stresses, i.e., water stress, fertilization, light, and salinity, were applied in two seasons (season 1: July to November and season 2: February to May). Bacoside A content in methanol extracts of the dried aerial parts of plants grown under different stresses was determined using a validated thin layer chromatography (TLC) densitometric method. In vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and Ellman’s method were employed to evaluate the antioxidant and acetylcholinestrase inhibitory activities of different extracts, respectively. Best plant growth was observed in soil A in season 1. Plants grown in season 2 had a significantly higher bacoside A content, better antioxidant, and acetylcholinestrase inhibitory activities than the plants grown in season 1. Among various stresses applied in season 2, plants grown in soil B, especially water stress-effected plants, had the highest bacoside A content, antioxidant, and acetylcholinestrase inhibitory activities. The plant should be cultivated from February to May in sandy loam soil with water stress for enhanced production of the marker compound and significant bioactivities.
  122. Tripathi, Pratibha and Tripathi, Arpita and Singh, Akanksha and Yadav, Vineet and Shanker, Karuna and Khare, Puja and Kalra, Alok, Differential Response of Two Endophytic Bacterial Strains Inoculation on Biochemical and Physiological Parameters of Bacopa Monnieri L. under Arsenic Stress Conditions, Journal of Hazardous Materials Advances, vol. 6, pp. 100055, May 2022. doi: 10.1016/j.hazadv.2022.100055.
    The accumulation of arsenic (As), in medicinal herbs, is detrimental due to its adverse effects on plant metabolism and consumer health. The present study aims to examine the ameliorative effect of two arsenate [As(V)] reducing endophytic bacterial strains (CIMAP-A4 and BAC-7) on As induced photo-toxicity in~Bacopa monnieri. The inoculation of BAC-7 and CIMAP-A4 in As contaminated soil showed an increase in biomass (26 and 84\%) and reduced As accumulation in plant tissue (42\% and 88\%) respectively. Strain CIMAP-A4 improved tolerance index, arsenate reductase activity, plant nutrient uptake (Zn, Mg, Ca), and non-protein thiol synthesis than BAC-7 under As stress.The indole acetic acid production and biofilm formation potential under As(V) stress (10 mg L−1) only in CIMAP-A4 could be the reason for the higher ameliorating effect. The strain CIMAP-A4 was identified as~Paenibacillus~sp. and could be a good option for As-induced stress mitigation in plants and successful bioremediation.
  123. Nichakool, Bunyaporn and Jamphon, A. and {Pootang-on}, Yupa and Techakriengkrai, Weerawan and Techakriengkrai, T., A Study about Brahmi (Bacopa Monnieri) Preparation Steps on Its Saponin Quantity, Trends in Sciences, 2021. doi: 10.48048/tis.2021.1439.
    Brahmi (Bacopa monnieri) is a medicinal herb containing bioactive compounds (Bacosides) in the saponin group that enhances memory and prevents dementia. Brahmi is not favored for cooking because it is very bitter. Nowadays, most people consume Brahmi as supplementary food, which makes it more expensive than consuming in food form, not getting fibers and other nutrients. Currently, there are only few studies that work to alleviate in Brahmi. Therefore, this study aimed to find out the saponin quantity in Brahmi after some pretreatments and the effect of these treatments on Bacosides and bitterness. Thus, suitable preparation steps for Brahmi have the highest remaining saponin quantity and are consumable. There were various techniques of preparation, depending on the concentration of salt used in crumpling and 1 time boiling of Brahmi to reduce its bitterness. The salt concentrations used in this study were 0, 10, and 20 \% (w/w) compared with the fresh herb. The saponin quantity was analyzed by high-performance liquid chromatography (HPLC). The results showed that the total amount of saponin in boiled Brahmi that crumpled with 0, 10, and 20 \% were significantly lower than fresh Brahmi (p {$<$} 0.05). Brahmi was crumpled with 10 \% salt before boiling had the highest total saponin quantity of 2.69±0.02 g/kg of fresh weight and the highest tasting scores. Thus, the preparation of crumpling with 10 \% salt before cooking, was suitable because consumers gained the highest saponin and accepted the taste of food containing Brahmi. HIGHLIGHTS Brahmi have many health benefits not only memory enhancer but also prevention of dementia People used Brahmi as supplementary food because it is very bitter and not favored for cooking Reduce the bitter with non chemical as crumpling with 10 \% salt concentration before cooking was highest saponin and accepted the taste of food containing Brahmi
  124. Silpa, S.G. and Smitha, G.R. and Sadananda, G.K. and Ranjitha, K. and Gowda, A.P. Mallikarjuna and Umesha, K., Effect of Drying and Packaging Methods on Physico-Chemical and Phytochemical Composition of {\emph{Brahmi}} ( {\emph{Bacopa Monnieri}} L.) with Respect to Shelf Life Enhancement, Medicinal Plants - International Journal of Phytomedicines and Related Industries, vol. 11, no. 1, pp. 73, 2019. doi: 10.5958/0975-6892.2019.00010.8.
    Brahmi (Bacopa monnieri L.) is a promising medicinal plant for the pharmaceutical and herbal industries due to its inclusive effect on central nervous system activity as a memory enhancer. Study was undertaken to assess the effect of different drying methods and packaging materials on physico-chemical properties of Brahmi. Fresh biomass of Brahmi was subjected to five different methods of drying viz., sun, solar tunnel, cabinet, low cost polyhouse and shade drying and four methods of packaging in gunny bags, polypropylene bags, cartons and HDPE. Among different drying methods, shade drying registered maximum drying time of around 15 days to reach the desired moisture content ({$<$}10\%) while, drying was rapid in cabinet drying method (12-15 hours) when dried at 50o±1oC. Maximum greenness (2.67), chlorophyll a (0.70 mg/g), chlorophyll b (0.26 mg/g) and total chlorophyll (0.96 mg/g) contents were retained in shade drying. Maximum bacoside A content (2.01\%) and texture (25.62 N) were recorded in tray dehydrated samples. Storage and packaging in polythene lined gunny bags and HDPE (500 guage) was found better in retaining the chlorophyll, color, texture, moisture and bacoside A content to a longer duration. The triterpene glycoside - bacoside A content of 2.02\% was best retained in cabinet dried material packed in HDPE even after one month of storage. Moisture content increased during storage and was found more pronounced in polypropylene bags and cartons. The shade dried sample stored in HDPE retained chlorophyll a content up to 0.54 mg/g after 5 months of storage which indicates the superiority of HDPE packaging over other methods. Among the different methods employed, cabinet drying or solar tunnel drying along with packaging in polythene lined gunny bags or HDPE were reflected as versatile methods of drying and packaging, respectively.
  125. Srivastava, Pratibha and Raut, Hema N. and Puntambekar, Hemalata M. and Desai, Anagha C., Stability Studies of Crude Plant Material of Bacopa Monnieri and Quantitative Determination of Bacopaside I and Bacoside A by HPLC, Phytochemical Analysis, vol. 23, no. 5, pp. 502--507, 2012. doi: 10.1002/pca.2347.
    Introduction Bacopa monnieri (BM) contains several dammarane-type triterpenoid saponins including bacopaside I and bacoside A. These bioactive compounds may be used as chemical markers for the quality control of different BM products used for promoting mental health and intellect. Objective Quantification of bacopaside I and bacoside A in crude plant material of BM stored under the stability study conditions by HPLC. Methodology Crude BM samples were stored at long-term (LS; 30°C and 65\% RH), accelerated (AS; 40°C and 75\% RH) and real-time (RT) study conditions. HPLC of BM extracts was carried out using a LiChroCART Purospher® STAR RP-18 endcapped column along with a guard column, Purospher STAR RP 18e 4.0 4.0 mm 5 µm using a gradient of acetonitrile (A) and water containing 0.05\% (v/v) orthophosphoric acid (B) at a flow rate 1.5 mL/min with UV detection at 205 nm. Results The linear range of bacopaside I and bacoside A was 0.2 to 1 mg/mL. With the help of a regression equation the coefficient of determination (r2) values for bacopaside I and bacoside A were found to be {$>$} 0.999 and {$>$} 0.994 respectively. Relative standard deviation (RSD) values were {$<$} 4.0 for all the concentrations injected (n = 3). The HPLC study indicated that BM samples kept under LS condition are rich in saponin contents as compared with the samples stored under AS and RT study conditions. Conclusion The study indicated that BM plant material should be used fresh to obtain maximum concentration of active saponins or it should be stored under LS conditions up to 3 months. Copyright © 2012 John Wiley \& Sons, Ltd.
  126. Panda, Debabrata and Barik, Jyoti R. and Barik, Jijnasa and Behera, Prafulla K. and Dash, Debasis, Suitability of Brahmi (Bacopa Monnieri L.) Cultivation on Fly Ash-Amended Soil for Better Growth and Oil Content, International Journal of Phytoremediation, vol. 23, no. 1, pp. 72--79, January 2021. doi: 10.1080/15226514.2020.1791052.
    Sustainable application of fly ash and its management in agriculture is a major challenge nowadays. A pot culture experiment was conducted to find out the most suitable level of fly ash application for soil amendments that can improve the plant growth and productivity of Brahmi (Bacopa monnieri L.). After growing seedlings of B. monnieri under different levels of fly ash for 90 days, a significant increase in plant biomass, essential oil content and tolerance index (more than 100\%) was observed under 25\% of fly ash amended soil in comparison to garden soil and higher fly ash treatments. Leaf chlorophyll content and photosynthetic parameters were remained unchanged under 25\% of fly ash as compared to seedlings grown on garden soil. However, these parameters were significantly declined under higher concentrations of fly ash treatments. Higher levels of fly ash caused oxidative damage and the induction of some antioxidative enzymes activities in B. monnieri indicates its capability to endure oxidative stress tolerance. Overall, our study showed that 25\% of fly ash can be used as soil amendment for cultivation of B. monnieri L. leading to enhance plant biomass and essential oil production.
  127. Smitha, G.R. and Kalaivanan, D. and Sujatha, S., Influence of Shade and Organic Nutrition on Growth, Yield and Quality of Memory Enhancing Herb, Brahmi (Bacopa Monnieri L.), Medicinal Plants - International Journal of Phytomedicines and Related Industries, vol. 13, no. 1, pp. 72--90, 2021. doi: 10.5958/0975-6892.2021.00009.5.
    Memory enhancing herb - Brahmi (Bacopa monnieri L.) has growing national and international demand owing to its various medicinal properties. In order to study the influence of shade and nutrient levels, an experiment was laid out in strip plot design with shade levels as the main plots (35\% shade, 50\% shade and 100\% sunlight) and nutrients requirement as sub plots [50, 75 and 100\% N equivalent through FYM, 50, 75 and 100\% N equivalent through vermicompost, RDF (100:60:60 kg NPK/ha), RDF (100:60:60 kg NPK/ha) + FYM (10 t/ha)]. The results revealed that among different shade levels tried, superior plant growth, fresh and dry herbage yield and cumulative dry herbage yield of seven harvests were obtained in 35\% shaded condition compared to 50\% shade and open conditions. Among different nutrient levels tried, conjunctive use of recommended dose of fertilizers (100:60:60 kg NPK/ha) + FYM (10 t /ha) gave the highest dry herbage yield (16.35 t/ ha), which was on par with organic treatment consisting of 100\% N equivalent through FYM + Arka microbial consortium @ 12.5 kg/ha/year (16 t/ha). However, highest content of bacopasaponin C and bacoside A were observed in 100\% sunlight condition (1.88 and 2.93\%, respectively) and in integrated nutrient management treatment. The results were very promising for acceptance of Brahmi for commercial cultivation especially in organic production scenario to ensure steady supply of quality raw material to cater the increasing national and international demand.
  128. Pankaj, Umesh and Singh, Durgesh Narain and Mishra, Pooja and Gaur, Pooja and Babu, C. S. Vivek and Shanker, Karuna and Verma, Rajesh Kumar, Autochthonous Halotolerant Plant Growth-Promoting Rhizobacteria Promote Bacoside A Yield of Bacopa Monnieri (L.) Nash and Phytoextraction of Salt-Affected Soil, Pedosphere, vol. 30, no. 5, pp. 671--683, October 2020. doi: 10.1016/S1002-0160(20)60029-7.
    Phytoremediation is a promising approach for reclamation of salt-affected soil. Phytoextraction is the most commonly used process, which exploits plants to absorb, immobilize, and accumulate salt in their shoots. In this study, halotolerant plant growth-promoting rhizobacteria (PGPR) were isolated from the rhizosphere of wild grasses growing naturally in salt-affected areas of Lucknow, Uttar Pradesh (India) and were tested for their efficacies of salt-tolerance and plant growth-promoting (PGP) abilities. Based on 16S rRNA sequences, the most efficient halotolerant isolates possessing PGP traits were identified as Pseudomonas plecoglossicida (KM233646), Acinetobacter calcoaceticus (KM233647), Bacillus flexus (KM233648), and Bacillus safensis (KM233652). Application of these isolates as bio-inoculants significantly (P {$<$} 0.05) increased the growth and bacoside A yield of a medicinal plant, Bacopa monnieri (L.) Nash, grown on natural salt-affected soil. The phytoremediation of salt-affected soil was evident by the substantial increase in shoot Na+:K+ ratio of bio-inoculant-treated plants. When compared to un-inoculated control plants, the soil physico-chemical properties of bio-inoculant-treated plants were improved. The shoot and root biomass (fresh and dry weights), soil enzymes, and soil nutrient parameters showed significant positive correlations with the shoot Na+:K+ ratio. Consequently, the halotolerant PGPR screened in this study could be useful for the reclamation of saline soils concomitant with improved plant growth and bacoside A yield.
  129. Maneeply, Chutiporn and Sujipuli, K. and Kunpratum, Narisa, Growth of Brahmi Bacopa Monnieri (L.) Wettst. by NFT and DFT Hydroponic Systems and Their Accumulation of Saponin Bacosides, NU. International Journal of Science, vol. 15, no. 2, pp. 114--124, 2018. url: https://www.sci.nu.ac.th/sciencejournal/index.php/journal/article/download/375/pdf.
    Brahmi or Bacopa monnieri (L.) Wettst. has been used as a nootropic herb in Ayurvedic medicine for thousands of years. This plant can boost memory retention and protect against depression and Alzheimer’s disease. The influence of NFT (Nutrient Film Technique) and DFT (Deep Flow Technique) hydroponic systems on growth of Brahmi and production of bacosides was investigated. Brahmi shoots were cultured in NFT and DFT hydroponic soilless systems for six weeks. NFT hydroponics grows plants using a very shallow recirculating nutrient solution, while DFT hydroponics employs a deep recirculating nutrient solution. Results showed that Brahmi biomass increased under hydroponic culture. The DFT technique stimulated vegetative growth of Brahmi more than the NFT technique, with the former generating higher dry weight, shoot number, leaf number and total leaf area. Highest dry weight of Brahmi plants grown under DFT hydroponics was 8.48±0.44 g/plant, 10.1 times higher compared to soil culture (control). Bioactive compounds in bacoside A consisting of bacoside A3, bacopaside II, bacoside X and bacopasaponin C were measured by high performance liquid chromatography (HPLC). High levels of bacoside A3, bacopaside II and bacoside X were observed in Brahmi grown in soil culture rather than by hydroponics, and Bacopasaponin C content (1.45±0.09\% w/w dry wt) was significantly higher than control (0.81±0.05\% w/w dry wt). Interestingly, accumulations of all active compounds of bacoside A per plant for Brahmi grown under hydroponics were significantly higher ( p \<0.01) than by soil culture. Findings indicated hydroponic cultures as an optimal alternative for Brahmi biomass production. Hydroponics gave a high-quality yield of Brahmi, high accumulation of bioactive compounds with fast crop production rate.
  130. Sharma, Munish and Khajuria, R.K. and Mallubhotla, Sharada, Annual Variation in Bacoside Content of Bacopa Monnieri (L.) Wettst Plants, International Journal of Pharma and Bio Sciences, vol. 4, pp. P266-P271, October 2013.
    A study was conducted to evaluate bacoside (expressed as sum of bacoside A3 and A2) production on a monthly basis in net house grown plants of Bacopa monnieri (accession BM001) collected from Jammu region. Bacoside content differed during the course of the whole year. The whole herb is used commercially for extracting the memory enhancing molecule-bacosides from the plant. The study significantly points out that the maximum bacoside yield was obtained during August to October (monsoon period) which also coincides with the favourable period for plant growth and would be suitable for plant harvest. Amongst the various individual plant parts, the highest content was recorded in the leaves (6.06 mg g-1 DW) followed by stem (5.13 mg g-1 DW) and least in the roots (3.19 mg g-1 DW).
  131. Sarnam, Singh and {Pandey C. S.} and Lal, Singh, Effect of Organic Manure and Nitrogen Fertilizer Application on Growth Rate, Nutrient Uptake and Yield of Jal Brahmi (Bacopa Monnieri Lenn.), Applied Biological Research, vol. 9, no. 1\&2, pp. 39--43, 2007. url: https://www.indianjournals.com/ijor.aspx?target=ijor:abr&volume=9&issue=1and2&article=007.
    A field experiment was conducted during 2002 and 2003 at CRC and MRDC, GBPUA\&T, Pantnagar to evaluate the effect of organic manure and nitrogen fertilizer on growth and nutrient uptake of Jal brahmi (Bacopa monnieri). The experiment was conducted on silty clay loam soil rich in organic carbon but medium in available phosphorus and potassium. The use of 75 kg N + 5 t farm yard manure (FYM) ha−1 resulted in maximum NPK uptake which was significantly higher than other treatments. Significantly higher mean crop growth rate was observed in 75 kg N + 5 t FYM ha−1 treatment, which yielded 53.4 and 45.5\% higher than control during 2002 and 2003, respectively. The application of 75 kg N + 5 t FYM ha−1 gave maximum number of leaves, branches spreading and yield which was significantly higher than all other treatments. Control plot recorded lower number of leaves, branches spreading and yield during both the years.
  132. Mathur, S. and Gupta, M. M. and Ram, M. and Sharma, S. and Kumar, S., Herb Yield and Bacoside-A Content of Field-Grown Bacopa Monnieri Accessions, Journal of Herbs, Spices \& Medicinal Plants, vol. 9, no. 1, pp. 11--18, February 2002. doi: 10.1300/J044v09n01_03.
    A field trial of five accessions of Bacopa monnieri (L.) Pennell was conducted during 1997 to 1998, at Lucknow, India, to standardize cultivation procedures for domestication of this medicinal herb. The accessions, monitored for growth and bacoside-A yields over 18 months, could be maintained as perennials, but growth properties were sensitive to the growing season. Loss of shoot biomass occurred in winter (December-February) and the growth rate was higher in the monsoon season (July-September) than in summer (March-June). Bacoside-A content of herb was high from September through March and in June. Suitable harvest times for high yields of bacoside-A were June and September through November. An accession from Guwahati in Assam state of India yielded more bacoside-A than all other accessions.
  133. Mathur, S. and Gupta, M. M. and Kumar, S., Expression of Growth and Bacoside-A in Response to Seasonal Variation in Bacopa Monnieri Accessions., Expression of growth and bacoside-A in response to seasonal variation in Bacopa monnieri accessions., vol. 22, no. 4a, pp. 320--326, 2000. url: https://www.cabdirect.org/cabdirect/abstract/20036792460.
    Wide variation in growth parameters as well as in bacoside-A content and yield was observed among 15 accessions of Bacopa monnieri collected from different geographical locations in India. In the plains of northern India, the accessions responded differently to the environmental conditions of winter (Sep.- Dec.), summer (Feb.- May) and monsoon (June - Sep.) seasons. Expression of vegetative...
  134. Bansal, Mahima and Reddy, M. Sudhakara and Kumar, Anil, Seasonal Variations in Harvest Index and Bacoside A Contents amongst Accessions of Bacopa Monnieri (L.) Wettst. Collected from Wild Populations, Physiology and Molecular Biology of Plants, vol. 22, no. 3, pp. 407--413, July 2016. doi: 10.1007/s12298-016-0366-y.
    Bacoside A, a major active principle of Bacopa monnieri known for its cognitive effects is a mixture of saponins like bacoside A3, bacopaside II, isomer of bacopasaponin C and bacopasaponin C. Seasonal changes in biomass and bacoside A levels in fourteen accessions of B. monnieri were evaluated after maintaining these at a common site at Thapar University campus, Patiala (30°19′36.12″N and 76°24′1.08″E) for 1~year. Harvestable biomass and total bacoside A contents varied significantly between the accessions and also in a particular accession during different seasons of the year. The maximum dry weight of plant (biomass 1.64~g) and bacoside A levels (6.82~mg/plant) were recorded in accession BM1. Harvestable biomass was highest during summer in accessions BM1 and BM7 (FW 4.2~g/plant), whereas bacoside A levels were also highest during summer and in accession BM1 (6.82~mg/plant). The lowest bacoside A level (0.06~mg/plant) was recorded in accession BM14 during winter. Principal component analysis showed that samples of summer were positively correlated with both the components suggesting an appropriate time for the harvest.
  135. Prasad, Ram and Kamal, Shwet and Sharma, Pradeep K. and Oelmüller, Ralf and Varma, Ajit, Root Endophyte Piriformospora Indica DSM 11827 Alters Plant Morphology, Enhances Biomass and Antioxidant Activity of Medicinal Plant Bacopa Monniera, Journal of Basic Microbiology, vol. 53, no. 12, pp. 1016--1024, 2013. doi: 10.1002/jobm.201200367.
    Unorganized collections and over exploitation of naturally occurring medicinal plant Bacopa monniera is leading to rapid depletion of germplasm and is posing a great threat to its survival in natural habitats. The species has already been listed in the list of highly threatened plants of India. This calls for micropropagation based multiplication of potential accessions and understanding of their mycorrhizal associations for obtaining plants with enhanced secondary metabolite contents. The co-cultivation of B. monniera with axenically cultivated root endophyte Piriformospora indica resulted in growth promotion, increase in bacoside content, antioxidant activity and nuclear hypertrophy of this medicinal plant.
  136. Kumar, Ardeep and Pandey, Sunita T. and Kumar, Ajit and Negi, M. S. and Gautam, Poonam, Innovation in Traditional Organic Nutrient Management Practices for Better Soil Health and Higher Productivity of Brahmi ( Bacopa Monnieri L.), Indian Journal of Traditional Knowledge (IJTK), vol. 21, no. 3, pp. 670--676, July 2022. doi: 10.56042/ijtk.v21i3.32007.
    This study was conducted at Medicinal Plants Research and Development Centre (MRDC) of Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, India, to examine the effect of Jeevamrit (Bioenhancer) on soil health and herbage yield of brahmi crop (var. CIM- Jagriti) and to optimise its rate of application. The experiment was laid out in randomised block design (RBD), replicated trice. The experimental soil was sandy clay loam in texture, neutral in reaction, having pH 6.9, medium in organic carbon (0.64\%), low in available nitrogen (180.78 kg ha-1) and medium in both available phosphorus (20.14 kg ha-1) and potassium (200.64 kg ha-1). Jeevamrit enhances soil nutrient content, improve bulk density and biodiversity by increasing beneficial soil microbes which mineralise the nutrients present in soil and increase their availability. Jeevamrit 4000 l ha-1 can be used as a nutrient source in place of chemical fertilisers and expensive bulky organic manures under organic nutrient management.
  137. Kumar, Udit, Evaluation of Brahmi (Bacopa Monnieri L.) Genotypes for Growth and Herbage Yield under North Bihar Agro-Ecological Conditions, Journal of Pharmacognosy and Phytochemistry, vol. 6, no. 6S, pp. 427--429, 2017. url: https://www.phytojournal.com/special-issue/2017.v6.i6S.2702/evaluation-of-brahmi-bacopa-monnieri-l-genotypes-for-growth-and-herbage-yield-under-north-bihar-agro-ecological-conditions.
    Brahmi, a member of Scrophulariacea family is a creeping, branched succulent perennial herb distributed in wet and marshy lands throughout Bihar. It is propagated by stem cuttings. The whole herb is the source of the ayurvedic drug ‘Brahmi’. It is used in improving memory and intelligence and also in the treatment of dermatosis, anaemia, diabetics and insanity. Bacoposide is considered as the major active ingredient in this plant. In the present study, 14(fourteen) collections (genotypes) of Brahmi, obtained from various places of Bihar were evaluated for their growth and herbage yield traits under north Bihar condition in the herbal garden under AICRP on Medicinal and Aromatic Plants \& Betelvine, Faculty of Agriculture, Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur, Bihar, during Kharif season of year 2015-16 to 2017-18. The pooled data of three years result revealed that there were significant variations among the genotypes with respect to vegetative growth and herbage yield. The variation among accessions for all seven characters was statistically significant. Maximum plant height, number of nodes per plant, number of primary branch per plant, leaf length and leaf width was recorded in accession no. IC-0622291. Maximum fresh herbage yield was recorded in the accession no. IC – 0622291 (222.53 q/ha) which was significantly superior to rest of the accession. Minimum was recorded in the accession no. IC – 0622293 (99.51q/ha). On the basis of growth and yield parameter it can be concluded that accession no. IC-0622291was the best genotype among the fourteen genotypes.
  138. Sujatha, S. and Bhat, Ravi and Kannan, C. and Balasimha, D., Impact of Intercropping of Medicinal and Aromatic Plants with Organic Farming Approach on Resource Use Efficiency in Arecanut (Areca Catechu L.) Plantation in India, Industrial Crops and Products, vol. 33, no. 1, pp. 78--83, January 2011. doi: 10.1016/j.indcrop.2010.09.001.
    The present investigation was conducted at Vittal, Karnataka, India during 2004–2007 to study the feasibility of intercropping of medicinal and aromatic plants (MAPs) in arecanut plantation. The results revealed that MAPs can be successfully grown as intercrops in arecanut plantation with increased productivity and net income per unit area. Kernel equivalent yield of MAPs varied between 272kgha−1 in case of Piper longum to 1218kgha−1 in Cymbopogon flexuosus. Pooled data indicated that Asparagus racemosus produced fresh root yield of 10,666kgha−1 of arecanut plantation and contributed to maximum kernel equivalent yield of 1524kgha−1 among all medicinal and aromatic plants. Intercropping of MAPs in arecanut was found economical. The net return per rupee investment was highest in C. flexuosus (4.25) followed by Bacopa monnieri (3.64), Ocimum basilicum (3.46) and Artemisia pallens (3.12). The total system productivity of arecanut+MAPs intercropping system varied from 2990 to 4144kgha−1. Arecanut+O. basilicum intercropping system registered significantly higher production efficiency 8.2kgha−1day−1 than other systems. Intercropping of MAPs had more positive effect on soil pH in arecanut based cropping system. The soil pH was 5.6 in 2004 and it was 0.3–0.9 units higher in 2007. Soil organic carbon (SOC) content varied significantly due to intercropping of MAPs at the end of experiment. The SOC content increased in Aloe vera, A. pallens, P. longum and B. monnieri, while it depleted in grasses and rhizomatic MAPs. Based on demand and marketing opportunities for MAPs, farmers are advised to grow aromatic plants in large areas on a community basis to meet huge industrial demand and variety of medicinal crops in small areas to meet the requirement of traditional systems of medicine.
  139. Pande, Preeti and Chand, Sukhmal and Yadav, Vinod K. and Anwar, M. and Patra, D. D., Influence of Chromium with Vermicompost on Growth and Accumulation by Brahmi, Communications in Soil Science and Plant Analysis, vol. 38, no. 19-20, pp. 2815--2829, November 2007. doi: 10.1080/00103620701663057.
    A pot experiment was conducted under glasshouse conditions during 2004 on brahmi (Bacopa monnieri) at the Central Institute of Medicinal and Aromatic Plants (CIMAP) in Lucknow. The study was conducted to evaluate the influence of different levels of chromium (Cr), with and without vermicompost, on growth and yield, as well as on accumulation of Cr by Bacopa. Rooted cuttings of Bacopa were grown under three levels of Cr (10, 20, and 40 ppm), two levels of vermicompost (2.5 and 5 g kg−1 soil), and a combination of both. The results indicated that herb yield increased with the joint application of vermicompost and chromium, as compared to compost and Cr alone. Application of Cr only decreased nitrogen (N) and increased phosphorus (P) concentration in plants with increase in its supply, whereas application with vermicompost raised N concentration. Chromium concentration in plant tissue was found to be highest at supply of Cr at 40 ppm+vermicompost 5 g kg−1 soil. Increase in the supply of Cr only (10, 20, and 40 ppm) decreased iron (Fe), copper (Cu), and zinc (Zn) concentration in plants. The experiment suggests that brahmi, because of its high accumulation ability, could be used as a scavenger to clean Cr‐contaminated soil.
  140. Krishantha, G and Gunasekera, H.K.L.K. and Sugathadasa, K and Prasangika, K, Feasibility Study of Growing Lunuwila (Bacopa Monnieri L.) Under Hydroponics, vol. 3, August 2022.
  141. Kaur, Puneet and Chhina, Gurbax and Kaur, Agumjot, Response of Brahmi (Bacopa Monnieri L.) Cultivars to Different Growing Media in Hydroponic System, Ecology, Environment and Conservation, pp. S215-S219, October 2022. doi: 10.53550/EEC.2022.v28i06s.036.
    The field experiment was conducted at Students’ Research Farm, Khalsa College, Amritsar during kharif2019-20. There were total of 10 treatments comprising two cultivars i.e. C1 – CIM-Jagriti and C2 – Subodhakand five growing media i.e. G1 – Cocopeat; G2 – Sawdust; G3 – Vermiculite; G4 – Mixture (cocopeat + sawdust+ vermiculite in 1:1:1) and G5 – Soil (control). The results revealed that higher growth and yield attributeswere found in cv. Subodhak than cv. CIM-Jagriti but they had non-significant difference whereas bacoside-A content was found to be higher in CIM-Jagriti than Subodhak. Growing media had influence on theperformance of brahmi plants. Brahmi grown in G1 responded better than other media. It was observedthat the growth and yield parameters of brahmi were significantly higher in G1 than G2, G3, G-4 and G5, butwas statistically at par with G-4. Plants grown in G5 showed significantly lower results than other media.
  142. Jain, Shilpi and Singh, Arjun and Mishra, Disha and Chanda, Debabrata and Khare, Puja, Reduction in Bioavailability of Pb in Bacopa Monnieri by Biochar Amendments, March 2015. doi: 10.13140/RG.2.1.4486.8647.
    Objective  The aim of the present study is to examine the effectiveness of biochar in reduction of phytoavailability of Pb in Bacopa monnieri. Acknowledgment The authors are thankful to the Director of CSIR-CIMAP for constant encouragement and help for this work. The financial support from project (BSC-203) and CSIR India is duly acknowledged. Conclusion  The present study reveals that Bacopa monnieri grown on biochar amended lead contaminated soil is safe for medicinal purpose. Graphical Abstract Introduction: Medicinal plants are currently used by large section of population. Evidence suggests that content of toxic heavy metals above permissible limits in these plant is a major concern for their consumption as they are freely available to everyone. Thus, it is an essential requirement to reduce the uptake of heavy metal during the cultivation. Biochar, a solid product of pyrolysis of biomass is gaining attention, because of its potential application to improve soil quality, sequester carbon and immobilize contaminants in soil. Objective of the study: The main goal of present study is to assess the bioavailability of Pb in Bacopa monnieri by application of different bio-chars. Material and Methods: The bioavailability of metal in B. monnieri grown in Pb contaminated soil was assessed using different biochars. The acute oral toxicity of plant extract was also performed for safety measure. Result and discussion: The results indicates that biochar not only reduce the bio-availability of Pb, but also improved the soil quality. The reduction of metal depends upon the biochar type and its properties. Safety evaluation showed that the plant extract had no adverse effect on body weight and metal accumulation in the major body organs and was found to be safe for use in traditional medicine. Conclusion: The present study reveals that Bacopa monnieri grown on biochar amended lead contaminated soil is safe for medicinal purpose. Abstract Higher yield of bacoside Reduced Phytotoxicity Biochar Pyrolysis
  143. Ak, Abdussalam and {chandra}, Ratheesh and Salim, Nabeesa, Heavy Metal Accumulation Potential and Medicinal Property of Bacopa Monnieri-a Paradox, journal of stress physiology and biochemistry, vol. 7, pp. 39--50, December 2011.
    Bioaccumulation of Mercury and Cadmium in Bacopa monnieri (L.) Pennell, cultivated in Hoagland medium artificially contaminated with micro quantities of HgCl 2 and CdCl 2 is investigated. Bioaccumulation potential of B. monnieri is more towards Cd than Hg. Absorption and translocation of Hg and Cd are proportional to the availability of the metal in the growth media and period of growth. Effect of acidic pH showed enhanced accumulation while basic pH resulted in significant reduction in the accumulation of Hg and exorbitant reduction of Cd. As a result of combined treatment of HgCl 2 and CdCl 2 , accumulation was very low in both acidic and basic pH. In addition to pH, antagonistic effect of Ca 2+ present in lime water which was added to raise the pH of the growth medium also controls the accumulation and absorption of Hg and Cd ions. Bioaccumulation of Hg and Cd in B. monnieri reveals the phytoremediation potential while the bioaccumulation is hazards to health since the plant is highly medicinal and one important ingredient of many Ayurvedic preparations.
  144. {Hussain-Koorimannil} and Abdussalam, A. K. and P, Ratheesh-Chandra and {Nabeesa-Salim}, Bio-Accumulation of Heavy Metals in Bacopa Monnieri (l.) Pennell Growing Under Different Habitat, International Journal of Ecology \& Development™, vol. 15, no. W10, pp. 66--73, January 2010. url: http://ceser.in/ceserp/index.php/ijed/article/view/366.
    Bio-accumulation of heavy metals was analysed in the samples of naturally growing Bacopa monnieri plants collected from eight different places/habitats of Kerala State. The heavy metals / elements such as Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn were present in considerable quantities which increased depending on the gravity of soil / water pollution due to geogenic and anthropogenic (industrial and pesticidal) effects. This bioaccumulation/phytoremediation property of Bacopa monnieri may lead to human health hazards because it is a widely used medicinal plant which grows profusely in wet lands flooded with industrial and sewage effluents.
  145. Joshi, Neeshu and Pandey, Sunita T. and Singh, V. Pratap and Jinger, Dinesh and Joshi, Shourabh and Paramesh, Venkatesh and Parihar, Manoj and Singhal, Rajesh and Javed, Talha and Saud, Shah and Hassan, Shah and Wang, Depeng and Wu, Chao and Fahad, Shah, Direct-Seeded Rice\,+\,Brahmi (Bacopa Monnieri) Intercropping and Weed Management Practices Affects Weed Control Efficiency and Competitive Indices, International Journal of Plant Production, November 2022. doi: 10.1007/s42106-022-00222-3.
    Weed menace is the major biotic constraint in direct-seeded rice (DSR) cultivation as they cause higher yield loss. Hence, exploring different weed management options along with intercropping could be promising practice for improving crop productivity. Keeping this in view, an experiment was conducted at Pantnagar, Uttarakhand, India in 2015 and 2016 to evaluate the effect of different weed management practices in the DSR\,+\,Brahmi intercropping system for providing farmers with a practical and economically viable weed control solution. Two intercropping ratios along with four weed management practices in sole rice and sole Brahmi as control were evaluated. The results revealed the significant effect of weed management and intercropping on weed control efficiency (WCE) and different competition indices. The highest WCE (73.3 and 83.3\%) was recorded with pendimethalin (pre-emergence)\,+\,cyhalofop-butyl (post-emergence)\,+\,1 HW (hand weeding) at 45 DAS (days after sowing) in 2:1 ratio treatment. Similarly, significantly minimum density and weed biomass were also recorded under the same treatment. However, significantly higher values of relative yield total, aggressivity, relative crowding coefficient, and competition ratio were recorded in a 1:1 ratio of DSR\,+\,Brahmi. In conclusion, using Brahmi as an intercrop with DSR can help with weed management, reduces the use of pesticides, and increases profitability.
  146. {Lastiri-Hernández}, Marcos A. and {Álvarez-Bernal}, Dioselina and {Ochoa-Estrada}, Salvador and {Contreras-Ramos}, Silvia Maribel, Potential of Bacopa Monnieri (L.) Wettst and Sesuvium Verrucosum Raf. as an Agronomic Management Alternative to Recover the Productivity of Saline Soils, International Journal of Phytoremediation, vol. 22, no. 4, pp. 343--352, March 2020. doi: 10.1080/15226514.2019.1663484.
    Water scarcity and increasing salinity are the main limiting environmental factors directly affecting the establishment and development of agroecosystems. The objective of this study was to assess the capacity of the halophytes Bacopa monnieri (L.) Wettst and Sesuvium verrucosum Raf., to improve the chemical properties of a saline soil during a 240-day period in the field so as to subsequently examine their potential to associate with Zea mays in the previously desalinated surface. The treatments proposed were [T1 (reference soil sample), T2 (soil + B. monnieri), T3 (soil + S. verrucosum), and T4 (soil + B. monnieri + S. verrucosum)]. The results showed that the association of the species B monnieri and S. verrucosum has the potential to enhance the chemical characteristics of the severely saline and clayey soil, showing ECe reductions of 11.13–7.97 dS/m and pH of 7.84–7.42, as well as increase in soil porosity from 54.71\% to 57.23\%. It was also found that the association of these plants have a phytodesalination capacity of 1.21 t Na+ ha−1, this served to prepare the conditions for the growth of the Z. mays and generate yields of 8.5 t ha−1.
  147. Patel, Anju and Jaiswal, Neha and Srivastava, Pankaj K. and Patra, D. D., Enhancing Secondary Metabolite Production and Antioxidants in Bacopa Monnieri Grown on Tannery Sludge Contaminated Soil, Industrial Crops and Products, vol. 187, pp. 115365, November 2022. doi: 10.1016/j.indcrop.2022.115365.
    The current study focuses on the stimulation of Bacoside A content of the Bacopa plant that played a potential role in the treatment of Alzheimer’s disease and epilepsy. The objective of the study was to explore the potential of the B. monnieri plant for metal tolerance when cultivated on tannery sludge-contaminated soil without compromising its phytochemical quality. Bacoside content of the plant gets 2 times higher than the control without any chemical additives. Bacosides are the alleged bioactive component and attribute to the neuroprotective function. Export-import bank of India acknowledged Bacopa monnieri as the second most crucial medicine as it has metabolites that can cure mental disorders and amnesia. The effect of tannery sludge on enzymatic and non-enzymatic antioxidant potential along with the effect on metabolite i.e. bacoside A content of the plant. The maximum transfer (known as translocation factor) from root to shoot for Cr, Cd, Co, Ni and Pb was 0.33, 0.28, 0.75, 0.69 and 0.43 respectively. This depicts that here in tannery sludge mixed soil, B. monnieri was not behaving like a hyperaccumulator plant. Although it sequestered a large portion of metal in the root region and showed high vegetative growth in a combination of 75:25 tannery sludge and soil. Superoxide dismutase, Catalase, peroxidase, malondialdehyde, proline \& ascorbic acid scaled up with increasing tannery sludge in the treatment whereas in the case of chlorophyll a, b, and carotenoid content, a sharp decline trend was observed at 100\% tannery sludge treatment. A similar trend was recorded in bacoside A content of the plant, and the highest was 2\% that of control in a 75:25 ratio of tannery sludge and soil. Enzymatic antioxidant along with non-enzymatic antioxidants protects the plant from oxidative stress generated by tannery sludge. Principal components confirm our finding that the translocation factor plays a direct role in the activation of the photosynthetic machinery. Principal component-I contributes 59\% of the total variance and Principal component-II contributes 33\% of the total variance.
  148. Adishesha, Veena and Murthy, K.M. Srinivasa, Phytochemical and Antioxidant Studies of Bacopa Monnieri Treated with Arbuscular Mycorrhizae Fungi Glomus Fasciculatum and Glomus Mosseae., Indian Journal of Applied Research, vol. 11, no. 7, pp. 47--52, July 2021. doi: 10.36106/ijar/9416618.
    Vesicular arbuscular mycorrhizae (VAM) play a crucial role in enhancing the growth of plant by increasing the uptake of nutrients from soil and also is known to enhance the secondary metabolites of plants. The response of VAM Glomus fasciculatumand Glomus mosseaeon the growth, phytochemical constituents and antioxidant properties ofBacopa monniericommonly known as Brahmi was carried out.B.monnieriwas treated with both pure cultures individually and mixed culture in (1:1) ratio along with uninoculated sample as control and gown in polyhouse condition for varying intervals. The physiological parameters, phytochemical analysis and antioxidant scavenging activities were analysed after 30, 60 and 90 days of VAM inoculation, respectively. Colonization of VAM was seen in the plant samples signifying the symbiotic association. Physiological analysis showed an increase in shoot, root and wet weight of B. monnieriwith increase in time. The preliminary phytochemical analysis inferred the methanolic leaf extracts of B.monnieritreated with Gf, Gm and Gf+Gmcontained alkaloids, avonoids, saponins, tannins, glycosides, terpenoids and carbohydrates which increased quantitatively with time.DPPH free radical scavenging activity showed escalation in result with increase in time of treatment with G. fasciculatumshowinghigher scavenging activity compared to Glomus mosseae and the mixed cultures.
  149. Mishra, Amrita and Mishra, Arun K. and Tiwari, Om Prakash and Jha, Shivesh, Studies on Metals and Pesticide Content in Some Ayurvedic Formulations Containing Bacopa Monnieri L., Journal of Integrative Medicine, vol. 14, no. 1, pp. 44--50, January 2016. doi: 10.1016/S2095-4964(16)60241-8.
    Objective Ayurvedic formulations are preferred over other formulations as well as commercialized on broad level to treat various ailments. The World Health Organization has established certain guidelines for quality control of heavy metals and pesticide residues. Bacopa monnieri, a popular herb with immunomodulator and memory-enhancing properties is the chief constituent of several Ayurvedic formulations, which include Brahmi Vati (BV), Brahmi Ghrita (BG) and Saraswat Churna (SC), etc. In view of the World Health Organization guidelines, two products of each formulation from six different manufacturers were purchased from Ayurvedic Pharmacy, Bulanala-Varanasi, India for testing heavy metal and pesticide residue. Methods In the present study, all the formulations—BV, BG and SC—were selected for estimation of four heavy metals namely lead (Pb), cadmium (Cd), chromium (Cr) and nickel (Ni) by a plasma emission spectrophotometer. Organochlorine pesticidal residues were estimated for dichlorodiphenyl trichloroethane, isomers of hexachlorocyclohexane (HCH) and α-endosulfan, etc. in total 12 samples of test formulations containing Bacopa monnieri L. using gas chromatography technique. Results Out of 12 samples, Pb, Cd, Cr and Ni were present in all samples but below the permissible limit. Although atrazine, aldrin, dialdrin were in below detection limit, but other pesticides were detected in some samples as oxamyl, hexachlorocyclohexanes (α-HCH, β-HCH and γ-HCH), dichlorodiphenyl trichloroethane and dichlorodiphenyl dichloroethylene. Conclusion The presence of heavy metals in the formulations was low to cause toxicity. However evaluation of heavy metals and pesticide residue in every batch is necessary.
  150. Srikanth Lavu, Rama Venkata and Prasad, Majeti Narasimha Vara and Pratti, Varalakshmi Lalithya and Meißner, Ralph and Rinklebe, Jörg and Van De Wiele, Tom and Tack, Filip and Du Laing, Gijs, Trace Metals Accumulation in Bacopa Monnieri and Their Bioaccessibility, Planta medica, vol. 79, no. 12, pp. 1081--1083, August 2013. doi: 10.1055/s-0032-1328713.
    Bacopa monnieri is commonly known as "Brahmi" or "Water hyssop" and is a source of nootropic drugs. Aboveground parts of plant samples collected from peri-urban Indian areas were analysed for total trace metal concentrations. Subsequently, three samples with high concentrations of Cd and Pb were subjected to in vitro gastrointestinal digestion to assess the bioaccessibility of the trace metals in these plants. The total concentrations of trace metals on a dry weight basis were 1.3 to 6.7 mg·kg⁻¹ Cd, 1.5 to 22 mg·kg⁻¹ Pb, 36 to 237 mg·kg⁻¹ Cu, and 78 to 186 mg·kg⁻¹ Zn. The majority of Bacopa monnieri samples exceeded threshold limits of Cd, Pb, Cu, and Zn for use as raw medicinal plant material or direct consumption. Therefore, it is necessary to evaluate Bacopa monnieri collected in nature for their trace metal content prior to human consumption and preparation of herbal formulations.
  151. Behera, Shradhanjali and Mallick, B. and Tiwari, T.N and Mishra, P., Investigation of Self-Neutralization of Acid Rain-Induced Acidifying Materials in Bacopa Monnieri, Adv. Sci. Lett., vol. 8, pp. 867--875, January 2014.
    ABSTRACT ALL living things are affected directly or indirectly by acid rain (AR). Biological effects of acid deposition due to AR on plants are numerous and complex. Usually, AR induced acidity is observed in plants and aquatic environment. It is a serious environmental problem. The original quality of the medium can be restoring by the introduction of alkaline substances or acid (H2SO4) neutralizing chemicals such as limestone and CaCO3 or other salts of calcium and phosphorous. In our earlier report it is observed that, instead of increasing acidity Bacopa monnieri herb shows a decrease in acidity due to sulfuric-simulated acid rain (S-SiAR) induction. Energy dispersive spectrometry and X-ray diffraction results confirm the presence of material phases and elements present in the plant materials. Analysis by both the X-ray techniques confirmed the presence of Ca and P elements and their phases such as (K2Ca(SO4)2:H2O) and (FeAl2(PO4)2(OH)2:8H2O) in the B. monnieri herb. So, it is expected that the chemical reactions of AR with the above material phases caused neutralization of H2SO4.
  152. Hussain, K, Bacopa Monnieri (l.) Pennell –a Good Biomarker of Water Pollution/Contamination, Journal of Stress Physiology and Biochemistry, vol. 6, no. 3, pp. 91--101, 2010. url: http://www.jspb.ru/issues/2010/N3/JSPB_2010_3_91-101.html.
    Effect of water pollution on Bacopa monnieri was studied by culturing their rooted propagules in various polluted water samples and Hoagland nutrient medium artificially contaminated with different micro-level concentrations of HgCl2. Anatomical observations of those plants showed safranin-stained masses deposited in the xylem vessels of stem. The plants treated in chemical solutions which are free from metallic ions, under threshold level of HgCl2, and control plants were devoid of such deposits. Similar deposits were observed in plants cultured in various local water samples. Atomic Absorption Spectrophotometric analyses of these water samples and the bioaccumulation property of the plant detected the presence of Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn at various levels. The occurrence of the localized stained deposits in the xylem vessels of the stem of the plants cultured in polluted/contaminated aqueous medium, eventhough the growth medium contamination is micro-levels, is indicative of high sensitivity of Bacopa monnieri plants towards water pollution irrespective of the chemical nature of the pollutants. Although these stained deposits are not specific to any individual element that causes pollution, detection of water contamination is possible by observing the safranin-stained masses in the xylem vessels of this medicinal plant.
  153. {Tabassum-Abbasi} and Patnaik, Pratiksha and Abbasi, Shahid Abbas, Ability of Indian Pennywort Bacopa Monnieri (L.) Pennell in the Phytoremediation of Sewage (Greywater), Environmental Science and Pollution Research, vol. 27, no. 6, pp. 6078--6087, February 2020. doi: 10.1007/s11356-019-07259-4.
    The freely and abundantly available amphibious plant Indian pennywort Bacopa monnieri (L.) Pennell was able to phytoremediate sewage (greywater) quickly and substantially in SHEFROL® (“sheet flow root level”) bioreactors, achieving reductions in the levels of several indicator parameters: suspended solids, chemical oxygen demand, biological oxygen demand, nitrogen, phosphorus, zinc, copper, nickel, and manganese to the extents of about 90\%, 76–77\%, 80\%, 65\%, 55\%, 56\%, 42\%, and 41\%, respectively at hydraulic retention times of just 6~h. As these indicators of primary, secondary, and tertiary treatments were achieved simultaneously in a single reactor compartment, the system presented in this paper promises to be simple, rapid, and economical, in achieving significant treatment of greywater.
  154. Jain, Shilpi and Singh, Arjun and Khare, Puja and Chanda, D. and Mishra, Disha and Shanker, Karuna and Karak, Tanmoy, Toxicity Assessment of Bacopa Monnieri L. Grown in Biochar Amended Extremely Acidic Coal Mine Spoils, Ecological Engineering, vol. 108, pp. 211--219, November 2017. doi: 10.1016/j.ecoleng.2017.08.039.
    The addition of biochar in acidic mine spoils contaminated areas has been suggested for buffering and improving the fertility as well as the reduction in the metal bioavailability. This study evaluated the effect of biochar on reducing the heavy metal content in the Bacopa monnieri L. grown in the biochar amended acidic mine spoil and associated toxicological risks. The mine spoil (overburden) was treated with a biochar made from the fast pyrolysis of waste of Cymbopogon flexuosus at 450°C at application rates of 15\% and 20\%. Plant analysis (biomass, chlorophyll, protein and lipid peroxidises), metabolite content and metal levels (Al, Cd, Cr, Cu, Fe and Pb) in the Bacopa monnieri L. herb. Hazardous quotient and acute oral toxicity of plant and its extract cultivated was carried out. The results showed that biochar amendment significantly improved biomass of the plant as well as reduced the metal content in plant tissues. The concentrations of Al, Cd, Cr, Cu, Fe and Pb in the plant tissues were within the international permissible limits. The risk assessment for bacopa consumption showed that the hazard quotients values were lower than the threshold level for herb ({$<$}1). Application of biochar further reduced hazard quotients values the indicating safe to the consumers. In vivo toxicity also suggests that no toxic effect of bacopa extract on the albino mice. It is concluded that the application of biochar in acidic mine spoils can be not only useful for the plant growth, but also for reduction in the metal toxicity of Bacopa monnieri L., if consumed.
  155. Nigam, N. and Yadav, V. and Mishra, D. and Karak, T. and Khare, P., Biochar Amendment Alters the Relation between the Pb Distribution and Biological Activities in Soil, International Journal of Environmental Science and Technology, vol. 16, no. 12, pp. 8595--8606, December 2019. doi: 10.1007/s13762-019-02257-y.
    In the present investigation, the impact of biochar amendments on the Pb fractionation pattern and soil enzymatic activities in soil was evaluated. Two spiking concentrations of exogenous Pb, two application rates (2\% and 4\%) of biochar and three plants such as Withania somnifera, Andrographis paniculata and Bacopa monnieri were taken for the study. Three experiment sets were designed for each plant. Distribution of Pb (in acid soluble, reducible, oxidizable and residual fractions), physicochemical and biological properties (β-glucosidase, acidic and alkaline phosphatases, urease and microbial biomass) of soil were examined. Several chemometric techniques were applied to this huge data to evaluate the association of soil microbial activities with Pb fractionation. The results revealed that biochar amendments altered the fractionation of Pb by shifting from mobile fraction (acid soluble) to immobile fraction (oxidizable and residual). The correlation analysis suggested that this could be attributed to the increase in cation exchange capacity, organic carbon and available phosphorus content in the soil after biochar amendments. The results clearly demonstrated that biochar amendments in Pb-contaminated soil improved the enzymatic activities in the soil. The multidimensional scaling, correlation and sequential equation modeling imply that biochar has a shielding effect on soil microbial activity against Pb stress from each fraction of soil.
  156. Gupta, Rupali and Singh, Akanksha and Srivastava, Madhumita and Shanker, Karuna and Pandey, Rakesh, Plant-Microbe Interactions Endorse Growth by Uplifting Microbial Community Structure of Bacopa Monnieri Rhizosphere under Nematode Stress, Microbiological Research, vol. 218, pp. 87--96, January 2019. doi: 10.1016/j.micres.2018.10.006.
    The modification of rhizosphere microbial diversity and ecological processes are of rising interest as shifting in microbial community structure impacts the mutual role of host–microbe interactions. Nevertheless, the connection between host-microbial community diversity, their function under biotic stress in addition to their impact on plant performances is poorly understood. The study was designed with the aim to analyze the tripartite interactions among Chitiniphilus sp., Streptomyces sp. and their combination with indigenous rhizospheric microbial population of Bacopa monnieri for enhancing the plant growth and bacoside A content under Meloidogyne incognita stress. Overall, plants treated with the microbial combination recorded enhanced growth as illustrated by significantly higher biomass (2.0 fold), nitrogen uptake (1.8 fold) and bacoside A content (1.3 fold) along with biocontrol efficacy (58.5\%) under nematode infected field. The denaturing gradient gel electrophoresis (DGGE) fingerprints of 16S-rDNA revealed that microbial inoculations are major initiators of bacterial community structure in the plant rhizosphere. Additionally, the plants treated with microbial combination showed maximum diversity viz., Shannon’s (3.29), Margalef’s (4.21), and Simpson’s (0.96) indices. Likewise the metabolic profiling data also showed a significant variation among the diversity and evenness indices upon microbial application on the native microflora. We surmise that the application of beneficial microbes in combinational mode not only helped in improving the microbial community structure but also successfully enhanced plant and soil health under biotic stress.
  157. Gupta, Rupali and Singh, Akanksha and Srivastava, Madhumita and Singh, Vivek and Gupta, M. M. and Pandey, Rakesh, Microbial Modulation of Bacoside A Biosynthetic Pathway and Systemic Defense Mechanism in Bacopa Monnieri under Meloidogyne Incognita Stress, Scientific Reports, vol. 7, no. 1, pp. 41867, February 2017. doi: 10.1038/srep41867.
    Plant-associated beneficial microbes have been explored to fulfill the imperative function for plant health. However, their impact on the host secondary metabolite production and nematode disease management remains elusive. Our present work has shown that chitinolytic microbes viz., Chitiniphilus sp. MTN22 and Streptomyces sp. MTN14 singly as well as in combination modulated the biosynthetic pathway of bacoside A and systemic defense mechanism against Meloidogyne incognita in Bacopa monnieri. Interestingly, expression of bacoside biosynthetic pathway genes (3-Hydroxy-3-methylglutaryl coenzyme A reductase, mevalonate diphosphate decarboxylase, and squalene synthase) were upregulated in plants treated with the microbial combination in the presence as well as in absence of M. incognita stress. These microbes not only augmented bacoside A production (1.5 fold) but also strengthened host resistance via enhancement in chlorophyll a, defense enzymes and phenolic compounds like gallic acid, syringic acid, ferulic acid and cinnamic acid. Furthermore, elevated lignification and callose deposition in the microbial combination treated plants corroborate well with the above findings. Overall, the results provide novel insights into the underlying mechanisms of priming by beneficial microbes and underscore their capacity to trigger bacoside A production in B. monnieri under biotic stress.
  158. Dr, Misal and Av, Kharde and Vd, Deshmukh and Bk, Talekar, Elicitation of Bacoside Content Using Aspergillus Niger Filtrate in Bacopa Monnieri (Brahmi), undefined, 2020. url: https://www.semanticscholar.org/paper/Elicitation-of-bacoside-content-using-Aspergillus-Dr-Av/9f602c667c074a8fd7894a475aaab11711bdc44e.
    Investigation clearly indicated that the Aspergillus niger filtrate significantly influenced the Bacoside production in Brahmi, and this affected the bacoside content in Bacopa monnieri. Micropropagation technique is used for the elicitation of bacoside content in Bacopa monnieri. In vitro propagation was carried out on Murashige and Skoogs medium supplemented with 6-benzylaminopurine (1.1 µM) and Indole-3- butyric acid (0.30 µM) with 3\% sucrose for four weeks. For elicitation variable concentrations of Aspergillus niger filtrate (0, 0.5, 1.0, 1.5, 2.0 ml/L) were incorporated in MS basal medium for 7 days. Present investigation clearly indicated that the Aspergillus niger filtrate significantly influenced the Bacoside production in Brahmi. After seven days elicitation, maximum enhancement in Bacoside (1.62\% DW) was reported at lower concentrations of Aspergillus niger filtrate (0.5 ml/L) over control.
  159. Shanmugam, Laxmi and Ahire, Mahendra and Nikam, Tukaram, Bacopa Monnieri (L.) Pennell, a Potential Plant Species for Degradation of Textile Azo Dyes, Environmental Science and Pollution Research, vol. 27, no. 9, pp. 9349--9363, March 2020. doi: 10.1007/s11356-019-07430-x.
    The current study highlighted the phytoremediation potential of Bacopa monnieri (L.) Pennell for most commonly used azo dyes which are resistant to degradation. Fourteen azo dyes (reactive: 09; direct: 05) upon treatments up to 40~mg/L were decolorized in the range of 90 to 100\% after incubation of 2~weeks in in vitro and hydroponic cultures. No significant alteration in growth of B. monnieri was observed in the presence of dyes R. Magenta MB, R. Navy Blue M2R, Dt. Orange RS, Dt. T Blue GLL, Dt. Sky Blue FF alone, and together in the medium. However, at increasing concentrations (60–100~mg/L), the percent dye decolorization was declined and showed a toxic effect on plant growth. The chlorophyll content declined while membrane damage and osmolyte accumulation were increased in dye treated samples. The biological conversion of produced metabolites was analyzed using FTIR and GC-MS. Our results suggest that the intermediates of Dt. Blue GLL degradation consist L-Proline, N-valeryldecyl ester, 3,5 Di-tert-butyl-4-trimethylsiloxytoulene, and 1,2-benzenedicarboxylic acid, diisooctyl ester. The antioxidative and oxidative enzyme activities in roots and leaves were significantly higher in the presence of dyes over control indicate that these enzymes are involved in degradation of dyes. Percentage seed germination, shoot and root length of seedlings of legume, cereal, and oilseed crop was not affected, suggesting the compatible nature of the produced metabolites. Our results revealed the remarkable ability of Bacopa monnieri for long-term operations that lead to the practical application of phytoremediation in textile industries.
  160. Jasim, B. and Daya, P. S. and Sreelakshmi, K. S. and Sachidanandan, P. and Aswani, R. and Jyothis, Mathew and Radhakrishnan, E. K., Bacopaside N1 Biosynthetic Potential of Endophytic Aspergillus Sp. BmF 16 Isolated from Bacopa Monnieri, 3 Biotech, vol. 7, no. 3, pp. 210, June 2017. doi: 10.1007/s13205-017-0788-4.
    Chemically unique environment of endophytes makes them to have various adaptive mechanisms for survival. One of such mechanisms involves the production of pharmacologically significant plant-specific metabolites. In the present study, 26 endophytic fungi were isolated from stem of Bacopa monnieri (L.) Wettst. plants. All the isolates were screened for bacopaside production property by HPLC. Among these, the fungal isolate BmF 16 which was identified as Aspergillus sp. was confirmed for bacopaside N1 production (m/z 796) by LC–MS/MS analysis. As the extract of BMF16 used in the study was prepared from the fifth generation of culture, the obtained result can be confirmed as due to fungal production of bacopaside. In addition, this property was identified only for one among the 26 fungi screened. As bacopaside N1 production in fungi has not yet been reported, the results of the study are novel.
  161. Vajpayee, Poornima and Dhawan, Alok and Shanker, Rishi, Evaluation of the Alkaline Comet Assay Conducted with the Wetlands Plant Bacopa Monnieri L. as a Model for Ecogenotoxicity Assessment, Environmental and Molecular Mutagenesis, vol. 47, no. 7, pp. 483--489, 2006. doi: 10.1002/em.20217.
    Wetlands play a key role in maintaining environmental quality, and wetlands plants could serve as model organisms for determining the genotoxic effects of pollutants contaminating these areas. In the present study, DNA damage was evaluated in a wetlands plant, Bacopa monnieri L., as a potential tool for the assessment of ecogenotoxicity. The Comet assay was used for detecting DNA damage in B. monnieri exposed to two model mutagens, ethyl methanesulfonate (EMS) and methyl methanesulfonate (MMS). Significant (P {$<$} 0.05) dose-dependent increases in DNA damage were observed following treatments conducted by exposing both isolated nuclei (acellular or in vitro exposure) and whole plants (in vivo exposure) to 0.01–5 mM EMS and 0.05–100 μM MMS for 2 hr at (26 ± 2)°C. The assay was then used to evaluate the genotoxic potential of cadmium (Cd), a wetlands contaminant. In vitro exposure of nuclei from untreated leaves to 0.001–200 μM Cd for 2 hr resulted in significant (P {$<$} 0.05) levels of DNA damage. Cd concentrations ≥0.01 μM induced DNA damage as evidenced by increases in the Olive tail moment. In vivo exposure of plants to 0.01–500 μM Cd for 2, 4, and 18 hr resulted in dose- and time-dependent increases in DNA damage in the nuclei isolated from roots and leaves. Cd-induced DNA damage was greater in roots than leaves. To our knowledge, this is the first report describing the use of a wetlands plant for genotoxicity assessment, using the Comet assay. Environ. Mol. Mutagen., 2006. © 2006 Wiley-Liss, Inc.
  162. Pierce, S. C. and Pezeshki, S. R. and Larsen, D. and Moore, M. T., Hydrology and Species-Specific Effects of Bacopa Monnieri and Leersia Oryzoides on Soil and Water Chemistry, Ecohydrology, vol. 2, no. 3, pp. 279--286, 2009. doi: 10.1002/eco.54.
    In an 8-week greenhouse experiment, Bacopa monnieri (water hyssop) and Leersia oryzoides (rice cutgrass) were compared for nutrient assimilation as well as soil and water chemistry under variable flooding regimes using a nutrient solution rich in nitrogen (N) and phosphorus (P). Soil redox potential decreased in flooded treatments; however, mesocosms containing B. monnieri remained aerobic for much of the study, while flooded mesocosms containing L. oryzoides became moderately reduced. Soils containing L. oryzoides were higher in nitrogen. Generally, effluent concentrations of PO43− were higher in B. monnieri mesocosms. B. monnieri immobilization of N and P was significantly less in below-ground tissues than L. oryzoides. P immobilization in L. oryzoides generally increased in response to flooding, while B. monnieri showed no detectable response. Results indicated that species-specific flood responses in plant nutrient status are due to differing interactions of B. monnieri and L. oryzoides with the soil environment. Additionally, L. oryzoides demonstrated greater P uptake than B. monnieri across treatments, resulting in decreased concentrations of PO4−3 in effluent. Although N was also affected by flooding and species, generalizations on N allocation within the system are difficult to describe due to the changes in species of N in response to oxidation–reduction gradients and biotic assimilation. Copyright © 2009 John Wiley \& Sons, Ltd.
  163. Suman, Shikha, Morphological Variations in Bacopa Monnieri, International Journal of Genetic Engineering and Recombination, vol. 1, no. 1, pp. 1--14, August 2015. doi: 10.37628/ijger.v1i1.25.
    Morphological Variations in Bacopa monnieri
  164. Roshni, L.s and Appukuttannair, Gangaprasad and Elenjikkal, Siril, Evaluation of Variability in Bacopa Monnieri (L.) Pennell Using Morphological and Biochemical Markers, International Journal of Applied Research in Natural Products, vol. 7, pp. 25--31, May 2014.
    Bacopa monnieri (L.) Pennell, is a well recognized medicinal herb having memory boosting property. Eleven accessions were collected from various parts of Kerala State, India and variation in thirty characters including twelve qualitative and eighteen quantitative traits were recorded. One way ANOVA on quantitative traits showed significant variations among accessions. The dendrogram generated on the basis of metric traits revealed accession BM 9 from Kozhikkode as the outlier. Principal component analysis (PCA) was performed to identify the significant characters to delimit the taxa and it revealed that quantitative characters like plant length, number of nodes per plant, inter nodal length, leaf length, pedicel length and flower length were the most significant ones. Biochemical characters including total saponin and bacoside A content were also found as the highly loaded characters. The variability records in B. monnieri can be implied to screen superior germplasm on the basis of desirable quality traits such as biomass, saponin and bacoside content. Industrial relevance. In the traditional, Indian system of medicine, several herbs have been used in the formulation of brain or nerve tonics. Bacopa monnieri is the prime species for this purpose. The plant is having a wide range of distribution pattern ranging from lower mild salt affected coastal belt to higher elevation in Kerala state. The plant is now cultivated as valuable raw drug resource in the state. However, the price of the raw drug is determined on the basis of quality, especially on the basis of its active principle. In the present report, morphological and biochemical traits (total saponin and bacoside A) including quality parameters of the raw drug excisable from B. monnieri were analyzed for accessions from different agro climatic regions in the state which can be used as a model for the screening of superior accessions, thus make organised cultivation of B. monnieri a profitable venture.
  165. Gubbannavar, Jyoti S, A Comparative Study of Roots of Bacopa Monnieri (l.) Pennel and Bacopa Floribunda (R.Br.)Wettst., no. 1, pp. 4, 2012.
    Bacopa monnieri a well known herbal drug , extensively using as memory enhancer all over the world. In Ayurveda Bhavaprakash nighantu described Brahmi(Bacopa monnieri) as Medhya, Ayushya, Rasayana. In Rajanighantu as Vatastra pittajit, buddhi, prajna, Medhya , Ayushyavardhini .Still utilizing from thousands of years due to lack of knowledge \& availability, improper identification we are using Bacopa floribunda and other species in the name of Bacopa monnieri. Present study differentiated Bacopa monnieri \& Bacopa floribunda in morphology, root TS, powder microscopy. Morphological character shows colour difference in flower, stem, root \& leaf, also shows difference in microscopic study and also in powder studies.
  166. Saesong, Tongchai and Temkitthawon, Prapapan and Nangngam, Pranee and Ingkaninan, Kornkanok, Pharmacognostic and Physico-Chemical Investigations of the Aerial Part of Bacopa Monnieri (L.) Wettst., pp. 8, 2019.
    A macroscopic, microscopic, physico-chemical, and phytochemical analysis of the aerial part of Bacopa monnieri (L.) Wettst. or Brahmi was undertaken for the purpose of establishing a monograph. The macroscopic and microscopic studies presented the typical characteristics of the Brahmi plant and its powder. The standard values of physico-chemical parameters were established at not more than 10.0 \%w/w for loss on drying and 13.0 \%w/w for total ash as well as not less than 17.0 \%w/w for water soluble extractives and 10. 0 \%w/w for ethanol soluble extractives. Phytochemical analysis showed the presence of steroidal saponins, with confirmation by the TLC fingerprint. Total saponin content was not less than 1.0 \%w/w measured by HPLC-UV method. The overall study provides pharmacognostical, physico-chemical, phytochemical details, typical TLC and HPLC fingerprints of the Brahmi, which can be used to establish a Pharmacopoeia monograph for the identification and standardization of Brahmi material.
  167. Bansal, Mahima and Kumar, Anil and Sudhakara Reddy, M., Diversity among Wild Accessions of Bacopa Monnieri (L.) Wettst. and Their Morphogenetic Potential, Acta Physiologiae Plantarum, vol. 36, no. 5, pp. 1177--1186, May 2014. doi: 10.1007/s11738-014-1493-0.
    Biochemical and molecular diversity among 14 accessions of Bacopa monnieri (L.) Wettst. collected from various locations of India was investigated. A significant variation was recorded in bacoside A contents of these accessions. A scatter plot of principle component analysis based on bacoside A contents clubbed these populations into two major groups and accession BM14 was placed separately. Similarly, about 35~\% variations were detected in these populations based on combined data of random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR). Individually, ISSR markers detected higher variation (44.9~\%) as compared to RAPD markers (23~\%). Clustering based on molecular marker data grouped these accessions into two major groups and also placed accession BM14 as an out group. The shoot organogenic potential of leaf explants taken from microshoots and rooting of microshoots also varied among accessions. Maximum shoot organogenic potential was observed in accession BM5 and maximum rooting potential was observed in accessions BM1, BM2, BM7, BM10 and BM14. Present study is an important step for developing long-term strategy for conservation of this important medicinal herb.
  168. Gubbannavar, Jyoti S. and Chandola, H. M. and Harisha, C. R. and Khanpara, Komal and Shukla, V. J., A Comparative Pharmacognostical and Preliminary Physico-Chemical Analysis of Stem and Leaf of Bacopa Monnieri (L.) Pennel and Bacopa Floribunda (R.BR.) Wettst, Ayu, vol. 34, no. 1, pp. 95--102, 2013. doi: 10.4103/0974-8520.115441.
    Brahmi is a well-known herbal drug having an effect on brain as a memory enhancer. Bacopa monnieri (L.) Pennel and Bacopa floribunda (R.Br.) Wettst are both marketed in the name of Brahmi. The present study differentiates Bacopa monnieri and Bacopa floribunda in morphology, transverse section (T.S.) of root and leaf, powder microscopy, and chemical constituents. Morphological characters show color difference in flower, stem and leaf and differences in microscopic study, organoleptic study, and powder characteristics. Morphologically, Bacopa monnieri leaf is fleshy and more succulent than Bacopa floribunda leaf. There is also a difference in the interval of the stem internodes of the two. Physico-chemical analysis revealed presence of 26\% bacoside A in Bacopa floribunda leaf and 27\% in Bacopa floribunda stem, which is higher than the bacoside A content in leaf (22\%) and stem (18\%) of Bacopa monnieri. However due to the hemolytic action of bacoside A, which is the toxic effect of the chemical constituent, it seems likely that Bacopa monnieri is more popular in regular use than Bacopa floribunda.
  169. Naik, P. M. and Manohar, S. H. and Praveen, N. and Upadhya, Vinayak and Murthy, H. N., Evaluation of Bacoside A Content in Different Accessions and Various Organs of Bacopa Monnieri (L.) Wettst, Journal of Herbs, Spices \& Medicinal Plants, vol. 18, no. 4, pp. 387--395, November 2012. doi: 10.1080/10496475.2012.725456.
    Bacoside A, a secondary metabolite in Bacopa monnieri, is used in the treatment of neuropharmacological disorders. Three phenotypic characters and bacoside A maker were assessed in 22 accessions of B. monnieri collected from Karnataka, India. Bacoside was highest (18.36 mg.g-1 DW) in accession collected from Belgaum (accession number Bm2) and lowest (3.53 mg.g-1 DW) in accession collected from Dharwad (accession number Bm7). Analysis of bacoside A in various parts of B. monnieri revealed variability and was highest in stolons (9.54 mg.g-1 DW) and lowest in leaves (1.91 mg.g-1 DW).
  170. Lal, Shalini and Baraik, Binit, Phytochemical and Pharmacological Profile of Bacopa Monnieri-an Ethnomedicinal Plant, 2019. url: https://www.semanticscholar.org/paper/PHYTOCHEMICAL-AND-PHARMACOLOGICAL-PROFILE-OF-BACOPA-Lal-Baraik/81cb1a9f546409ac7eee97433df47c7bb22307a0.
    The phytoconstituents, traditional uses, and pharmacological activities of Bacopa monnieri are described, which are an important medicinal plant of the family Scrophulariaceae used in traditional medicine to treat various nervous disorders and for promoting memory and intellect. In recent times, the use of herbal products has increased tremendously in the developing as well as in developed countries. Bacopa monnieri is one of the most important medicinal plant that has been consumed for thousands of year as a memory enhancing, anti-inflammatory, analgesic, antipyretic, sedative and anti-epileptic agent. For many years, a lot of commercial and medicinal researches have been focusing their resources on this plant. The plant has been known to possess various activities such as, antibacterial, anti-fungal, anti-cancer, anti-oxidant, anti-inflammatory, antihyperglycemic, anti-depressant, anti-epileptic, memory enhancer, anti-ulcer, hepatoprotective, analgesic, anti-diarrheal, anti-hypertensive, anti-toxicity etc. Therefore step should be taken for its sustainable use and conservation. This review describes the phytoconstituents, traditional uses, and pharmacological activities of Bacopa monnieri. In brief, the biotechnological applications such as tissue culture, elicitation, genetic transformation for the advancement of this plant and its active components production have also been described. INTRODUCTION: In human society from time immemorial medicinal plants have played an important role in the prevention and control of diseases. It has been confirmed by WHO that herbal medicines serve the health needs of about 80 percent of the world\&\#39;s population; especially for millions of people in the vast rural areas of developing countries. Development of science and technology and the side effect of modern medicine has resulted in increased and effective usage of plant-based medicines. Awareness of medicinal plants usage is a result of the many years of struggles against illnesses due to which man learned to pursue drugs in barks, seeds, fruit bodies, and other parts of the plants. QUICK RESPONSE CODE DOI: 10.13040/IJPSR.0975-8232.10(3).1001-13 The article can be accessed online on www.ijpsr.com DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.10(3).1001-13 In almost all the traditional medical systems, the medicinal plants play a major role and constitute their backbone. India has a rich heritage of traditional medicine and traditional health care system. Bacopa monnieri L.: Bacopa monnieri is an important medicinal plant of the family Scrophulariaceae used in traditional medicine to treat various nervous disorders and for promoting memory and intellect. This medicinal plant is locally known as Brahmi. It is known as a memory enhancer, and many preparations of Brahmi are now commercially available in the market. Some important medicinal uses of the plant B. monnieri for treatment of different diseases and the traditional formulation are given in Table 1. Habitat, Geographical Distribution, and Ethnomedical Description: Bacopa monnieri is a perennial, creeping herb whose habitat includes wetlands and muddy shores. The leaves are succulent, relatively thick, oblanceolate and are
  171. Malini, Shanmugarajan and Eganathan, Palanisami, GC-MS Analysis of Chemical Composition of in Vivo Plant, in Vitro and Elicited Roots of Bacopa Monnieri (L.) Pennell, Analytical Chemistry Letters, vol. 3, no. 5-6, pp. 380--388, November 2013. doi: 10.1080/22297928.2013.797636.
    In vitro plant was developed by tissue culture utilizing nodal explants and adventitious roots were obtained from nodal explants of Bacopa monnieri in Murashige and Skoog (MS) medium supplemented with 0.2 mg/l-1 NAA. In vitro roots of Bacopa monnieri were subjected to chemical elicitation using 69 mg/l-1 of salicylic acid by hairy root culture. In vivo whole plant, in vitro and elicited roots were subjected to solvent extraction using ethanol and their chemical compounds analyzed using gas chromatography and mass spectroscopy (GC-MS). The major composition of in vivo plant were 9,12-octadecadienonic acid (36.96 \%), 9,17-octadecadienal (26.65 \%,), 9-octadecenoic acid (7.79 \%) and in vitro roots yielded 9,12-octadecadienonic acid (25.62 \%), 9-octadecenoic acid (23 \%), 9,17-octadecadienal (16.08 \%). In vitro roots subjected to salicylic elicitation comprised of 1,3-dihydroxyacetone dimmer (15.69 \%), 1-hexadecena (7.74 \%), 1-tetradecene (6.78 \%), eicosane (6.57\%), 1-octadecene (5.29 \%), 1-decene (4.60 \%), E-15-heptadecenal (4.45 \%) and heptacosane (3.45 \%). In vitro elicited roots showed thirty six compounds and increased higher percentage of sesquiterpenoids and higher alkens.
  172. Kawai, Ken-Ichi and Shibata, Shoji, Pseudojujubogenin, a New Sapogenin from Bacopa Monniera, Phytochemistry, vol. 17, no. 2, pp. 287--289, January 1978. doi: 10.1016/S0031-9422(00)94166-9.
    Acid hydrolysis of bacoside A, a saponin of Bacopa monniera, afforded ebelin lactone and bacogenin-A1, while Smith-de Mayo degradation of bacoside A yielded jujubogenin and pseudojujubogenin as sapogenins. The structure of pseudojujubogenin except the absolute configurations at C-20 and C-22 was established by chemical and spectroscopic investigations. Bacoside A was separated by droplet counter-current chromatography into two fractions, A1 and A2, which, however, are mixtures, since they both afforded two sapogenins, in different ratios by Smith-de Mayo degradation.
  173. Shaba Khan, Naureen and Chaurasia, Bhaskar and Kumar Dixit, Ashwini, Pharmacognostic Characterization for Taxonomic Identification of Bacopa Monnieri (L.) Wettst. for Quality Control, International Journal of pharma and Bio Sciences, vol. 11, no. 1, pp. 54--62, January 2021. doi: 10.22376/ijpbs/lpr.2021.11.1.L54-62.
  174. Srivastava, Anshu and Garg, Gaurav and Sharma, Poojadevi and Shah, Nancy and Sharma, Sonal and Shrivastava, Neeta, Genetic Diversity in Chemically Diverse Accessions of Bacopa Monnieri, JPC – Journal of Planar Chromatography – Modern TLC, vol. 29, no. 3, pp. 203--208, June 2016. doi: 10.1556/1006.2016.29.3.6.
    Bacopa monnieri (L.) Pennell, commonly known as Brahmi, is an important medicinal plant of the Ayurvedic System of Medicine and is categorized as Medhya Rasayana, used for boosting memory. Bacoside A and B (saponins) are major bioactive compounds of the plant responsible for memory enhancement. Consistent quality and quantity of the bioactive compounds are a must to produce highly efficacious herbal drugs. The phytochemical diversity due to ecogeographical variations affects the persistent therapeutic effect. In the present study, quantification of the major bioactive principle of B. monnieri, bacoside A, was performed in order to evaluate chemodiversity in the plants collected from different ecogeographical locations. The analysis of 75 accessions showed a wide range of significant variation in bacoside A content. Further, genetic diversity study was carried out between high- and low-yielding accessions to correlate the effect of the genetic composition on chemical diversity. The cluster analyses of amplified fragment length polymorphism (AFLP) data of high and low bacoside A producing plants using dendogram and principal coordinate analysis (PCoA) revealed no specific clustering. The genetic diversity parameters calculated also support less genetic differentiation between low- and high-yielding accessions. The results clearly indicate that there is a prevalence of agroclimatic conditions on differential bacoside A accumulation in B. monnieri. The study will help to produce quality plant material to be used as phytomedicine.
  175. kumar Maruthai, mohan and Ragupathi, Gopi and Lakshmanan, Ganapathy Murugan and Panneerselvam, Rajaram and Edwinraj, Esack, Studies on Genetic Diversity of Bacopa Monnieri (L.) Pennell Ecotypes Variants from South India by the RAPD Markers, Romanian Journal of Biology - Plant Biology, vol. 58, pp. 9--17, January 2013.
    Brahmi is a small prostrate herb that grows wild in marshy and damp places near water logs throughout India. The plant has been used since ancient time in folklore and traditional medicine as nervine, cardio-tonic, and diuretic. The plants were collected from18 different geographical regions of south India. RAPD profiles in order to assess diversity and the similarity matrices were generated from the RAPD data on the basis of Jaccard similarity coefficient estimates of similarity indices and dendrogram were constructed based on UPGMA clustering. OPL-05 having the maximum range of polymorphism was noticed (83.33\%) and high level of monomorphism was noticed in the primer OPT-18. The amplification range is between 100 bp to 2900 bp and the total percentage of polymorphism is (50\%) was observed. The observed narrow genetic base in B. monnieri population in Tamil Nadu may be attributed to the vegetative propagation of the species.
  176. Dey, Abhijit and Hazra, Alok Kumar and Nandy, Samapika and Kaur, Prabhjot and Pandey, Devendra Kumar, Selection of Elite Germplasms for Industrially Viable Medicinal Crop Bacopa Monnieri for Bacoside A Production: An HPTLC-Coupled Chemotaxonomic Study, Industrial Crops and Products, vol. 158, pp. 112975, December 2020. doi: 10.1016/j.indcrop.2020.112975.
    Background Bacopa monnieri (L.) Wettst., commonly known as Brahmi or water hyssop is an age old Ayurvedic nootropic and cognitive booster used as a medicinal food and also in an array of pharmaceutical and healthcare products as a memory enhancing tonic. It is a high value medicinal crop in modern healthcare industry owing to a number of bioactive pharmaceuticals of which bacoside A is of prime neuropharmacological importance. The bacosides-enriched standardized extract of Bacopa is being used and investigated for clinical purpose. Objectives The objectives of this study are to analyze the intrinsic intra-specific variation in bacoside A in B. monnieri populations, to identify the elite (high bacoside A yielding) chemotypes of B. monnieri, to investigate the in vitro acetylcholinesterase (AChE)-inhibitory activity of the B. monnieri elite chemotypes and to find out the cytotoxicity and genotoxicity of B. monnieri employing Allium cepa root tips. Material and methods: This present work describes a quick, validated and reproducible high-performance thin layer chromatography (HPTLC) method to estimate bacoside A content from 25 chemotypes of B. monnieri procured from 25 districts of 8 states of India. Further, an evidence-based approach was followed to validate the reported nootropic activity of the aqueous extracts of the plants by assessing their AChE inhibitory activity. Finally, the genotoxicity of an elite (high bacoside A producing) chemotype was assessed using Allium cepa root tip meristematic cells. Results B. monnieri chemotype BM1 (collected from North 24 Pgs, West Bengal) (7.55\,μg) was found to possess the highest amount of bacoside A followed by BM3 (6.43\,μg) and BM6 (6.02\,μg) chemotypes collected from Jalpaiguri, West Bengal and Ranchi, Jharkhand respectively. Chemotype BM 25 (2.17\,μg) exhibited least amount of bacoside A. Bacoside A content ranged between 2.17–7.67\,μg across 25 chemotypes of B. monnieri. The strongest AChE inhibitory activities (IC50 values of 259.45 and 267.89\,mg/mL respectively) were showed by the highest bacoside A containing chemotype BM1 (259.45\,mg/mL). Allium cepa root tip meristematic cell assay also revealed the lack of mitotoxic and cytotoxic property of the plant extract. Conclusion The present rapid, reproducible and validated HPTLC provides an efficient tool to determine the quantitative variation in the bacoside A content in naturalB. monnieri chemotypes. This method may be applied to select high bacoside A producing elite genotypes for further propagation and commercialization of this significant neuro protectant for pharmacological industry. Besides, it’s in vitro AChE inhibitory property directly supports the popular use of the plant as a nootropic agent. Lack of genotoxicity of the plant extracts evidences its age old use as a medicinal food and commercial memory booster.
  177. Christopher, Cinthya and Johnson, Anil John and Mathew, Paravanparampil Jacob and Baby, Sabulal, Elite Genotypes of Bacopa Monnieri, with High Contents of Bacoside A and Bacopaside I, from Southern Western Ghats in India, Industrial Crops and Products, vol. 98, pp. 76--81, April 2017. doi: 10.1016/j.indcrop.2017.01.018.
    Bacopa monnieri (Brahmi) is a high value herbaceous medicinal species used for improving intellect and memory. In India, high demand of B. monnieri for the manufacture of herbal extracts and medicines prompts its collection in huge quantities from wild habitats, leading to the extinction of its genotypes. Wild accessions of B. monnieri show high variability in the levels of bioactive principles, and only few of them yield ‘elite’ contents. The objective of the present study is search for elite genotypes of B. monnieri in its wild germplasm across the south Indian State of Kerala. A precise HPTLC-densitometric method was adopted for simultaneous quantification of the bioactive saponins, Bacoside A and Bacopaside I, in sixty accessions of B. monnieri originally collected from various locations in Kerala and grown under identical ecological conditions in a Field Gene Bank/Experimental Plot for four generations. These sixty B. monnieri accessions showed high variability in the contents of bioactive compounds, and three of them showed elite contents ({$>$}5.5\%) of (Bacoside A+Bacopaside I), viz., 6.58, 6.55 and 5.56\%. Total content of both bioactive constituents (Bacoside A+Bacopaside I) ranged from 2.06 to 6.58\%. Bacoside A and Bacopaside I contents in these sixty accessions varied from 1.44 to 5.40\% and 0.29 to 1.36\%, respectively. The average content of (Bacoside A+Bacopaside I) in these sixty accessions of B. monnieri is 3.70±0.72\%. Elite genotypes of B. monnieri discovered in this study are potential entities for cultivation and genetic improvement, which could ultimately meet its commercial requirements.
  178. Jeena, Gajendra Singh and Fatima, Shahnoor and Tripathi, Pragya and Upadhyay, Swati and Shukla, Rakesh Kumar, Comparative Transcriptome Analysis of Shoot and Root Tissue of Bacopa Monnieri Identifies Potential Genes Related to Triterpenoid Saponin Biosynthesis, BMC Genomics, vol. 18, no. 1, pp. 490, June 2017. doi: 10.1186/s12864-017-3865-5.
    Bacopa monnieri commonly known as Brahmi is utilized in Ayurveda to improve memory and many other human health benefits. Bacosides enriched standardized extract of Bacopa monnieri is being marketed as a memory enhancing agent. In spite of its well known pharmacological properties it is not much studied in terms of transcripts involved in biosynthetic pathway and its regulation that controls the secondary metabolic pathway in this plant. The aim of this study was to identify the potential transcripts and provide a framework of identified transcripts involved in bacosides production through transcriptome assembly.
  179. Sidharthan, V. Kavi and Baranwal, V. K., Mining of the Water Hyssop (Bacopa Monnieri) Transcriptome Reveals Genome Sequences of Two Putative Novel Rhabdoviruses and a Solendovirus, Archives of Virology, vol. 166, no. 7, pp. 1985--1990, July 2021. doi: 10.1007/s00705-021-05061-7.
    The genomes of three putative novel viruses, tentatively named "Bacopa monnieri virus 1" (BmV1), "Bacopa monnieri virus 2" (BmV2), and "Bacopa monnieri virus 3" (BmV3) were identified in the transcriptome dataset of a medicinally important herb – water hyssop (Bacopa monnieri (L.) Wettst.). The BmV1 and BmV2 genomes resemble those of plant rhabdoviruses. The 13.3-kb-long BmV1 genome contains eight antisense ORFs in the order 3′ l–N–P2’–P–P3–M–G–P6–L–t 5’, with P2’ ORF overlapping with P, while the 13.2-kb BmV2 genome contains six interspersed ORFs in the antisense orientation (3′ l–N–P–P3–M–G–L–t 5’). The 8-kb BmV3 genome possesses five overlapping ORFs, with ORFs 2 to 5 being similar to those of solendoviruses. Based on genome organization, sequence similarity, and phylogeny, BmV1, BmV2, and BmV3 can be regarded as new members of the genera Cytorhabdovirus, Betanucleorhabdovirus, and Solendovirus, respectively.
  180. Anu, Sanjoris and Remmies, Rinnu and Jose, Catherine, Phylogenetic Polymorphism in Neera-Brahmi, Bacopa Monnieri (L.) Using Molecular Markers, International Journal of Advances in Science, Engineering and Technology, vol. 5, no. 3, pp. 68--70, 2017. url: http://ijaseat.iraj.in/paper_detail.php?paper_id=9116&name=Phylogenetic_Polymorphism_in_Neera-Brahmi,_Bacopa_Monnieri_(L.)_using_Molecular_Markers.
    Bacopa monnieri (L.) Penn. commonly known as the Neera-Brahmi belongs to the family Scrophulariaceae is a small prostrate herb that grows wild in marshy and damp places near water logs spreading throughout India. Over exploitation of this herbal plant species and the necessity of biodiversity conservation of this endangered plant species have attracted global attention. Accordingly, among the seven highly endangered identified plant species that needed immediate attention, B.monnieri plants have a unique position as has been highlighted by NMPB, India and Technology Information Forecasting and Assessment Council (TIFAC), DST, Government of India (http://www.nmpb.nic.in/prioritisemedicinalplants.htm). In this way Biotechnologists also took much interest in analyzing and monitoring the systemic status of this valuable plant species.Identification of plant varieties/genotypes using modern molecular markers as PCR/RFLP profile have always been sought as reliable, inexpensive, easily obtainable and versatile set of molecular tools as repeatable DNA amplification sequence being relied up on in describing the accurate systematic status of any plant or animal species. The present study envisages to trace the genetic variations using their conserved genome like mitogenome and other different long and short primer polymorphic sequences thereby intend to explore the population structure of Bacopa monnieri (L.) in India. Keywords - Bacopa monnieri, Genetic variation, Inter simple sequence repeats (ISSR), Polymorphism, Random amplified polymorphic DNA (RAPD)
  181. Prabhudas, Sudheesh K. and Natarajan, Purushothaman, De Novo Assembly of Transcriptome and Draft Chloroplast Genome from RNAseq Data of Bacopa Monnieri L. (Bramhi), Canadian Journal of Biotechnology, vol. 1, no. Special Issue, pp. 193--193, 2017. doi: 10.24870/cjb.2017-a179.
    Medicinal plant naming services have recorded at least 28,187 plant species as being of medicinal use. Only 16\% (4,478) of these plants have been cited in medicinal regulatory publications and even a lesser proportion of it has ready-to-use transcript sequence information available in public data bases. Bacopa monnieri L. or Bramhi is a widely used medicinal herb mentioned in ancient ayurvedic scripts as a part of medhya rasayanas (brain rejuvenating neutraceuticals). In spite of being an extensively studied plant, it has very little genetic resources in pubic databases, thereby limiting extensive molecular studies based on genetics. In this study we sequenced the whole transcriptome of B. monnieri L. using Illumina Hiseq 2500 producing \textasciitilde 78 million high quality reads, followed by the de novo assembly generating a transcriptome size of \textasciitilde 88Mb and 111,290 clustered unigene transcripts. Plant non-redundant database, pfam and uniprot database were used as reference databases and a total of 59,260 (53.25\%) transcripts were annotated based on similarity searches. Pathway mapping of the unigenes using Kyoto encyclopedia of genes and genomes revealed 14,816 transcripts involved in 143 pathways. The triterpenoid and sesquiterpenoid biosynthesis pathway was selected to validate the assembled transcripts as the bioactive compounds in B. monnieri L. are reported to be triterpenoids. Simple sequence repeat (SSR) analysis excluding mono-nucleotide repeats showed the presence of 10,556 SSR's in a total of 8892 transcripts. An attempt was made to assemble the draft chloroplast genome from the assembled transcriptome data and \textasciitilde 65\% of chloroplast genome has been assembled and in progress. The draft chloroplast genome will also help in shedding light towards the evolution of B. monnieri L. The current study will provide information about key enzymes involved in various biosynthetic pathways and also a resource for comparative genomic and transcriptomic studies in future.
  182. Yadav, Aradhana and Ahmad, Javed and Chaudhary, Anis and Ahmad, Altaf, Development of Sequence Characterized Amplified Region (SCAR) Marker for the Authentication of Bacopa Monnieri (L.) Wettst, European Journal of Medicinal Plants, vol. 2, pp. 186-198., January 2012. doi: 10.9734/EJMP/2012/1192.
    Aims: To develop Sequence Characterized Amplified Region (SCAR) marker for identification of Bacopa monnieri (L.) Wettst. Study design: Molecular biology tools for authentic identification of Bacopa monnieri. Methodology: RAPD-based SCAR marker was developed to identify Bacopa monnieri from its adulterant candidates namely Centella asiatica, Eclipta alba and Malva rotundifolia. 50 random primers were used for initial screening of different accessions of Bacopa monnieri, Eclipta alba and Malva rotundifolia. A putative 589 bp marker specific to Bacopa monnieri was identified using RAPD technique. This RAPD-amplicon was then sequenced and cloned. Based on the information of cloned sequences a pair of SCAR primers was designed. SCAR primers were then used for authentication of DNA samples of Bacopa monnieri and its adulterants. Market samples of Bacopa monnieri and Centella asiatica collected under the name of Brahmi was put to test with these primers. Results: Out of 50 random primers, only 14 primers were able to amplify the above plants. A 589 bp polymorphic band obtained with OPAA-3 primer which was specific to Bacopa monnieri accessions and not found in other adulterant candidates was selected. This band was eluted, cloned and further sequenced. A pair of SCAR primers (Bac F \& Bac R) between 406 bp of 589 bp sequence of RAPD amplicon was designed. A single, bright, distinct band was obtained in Bacopa monnieri and not in the adulterants. Further validation was also done in the market samples. Conclusion: In essence, the study was to develop a RAPD-based SCAR marker for authentication of Bacopa monnieri. The SCAR marker was found to be useful for Research Article
  183. Tripathi, Niraj and Chouhan, Devendra Singh and Saini, Navinder and Tiwari, Sharad, Assessment of Genetic Variations among Highly Endangered Medicinal Plant Bacopa Monnieri (L.) from Central India Using RAPD and ISSR Analysis, 3 Biotech, vol. 2, no. 4, pp. 327--336, December 2012. doi: 10.1007/s13205-012-0059-3.
    Genetic variations of 15 Brahmi (Bacopa monnieri L.) accessions were evaluated using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers. During RAPD analysis, amplification of genomic DNA of the 15 accessions by 22 primers generated 197 fragments, of which 187 were polymorphic with an average of 8.95 bands per primer. The amplified products varied in size from 2,200 to 250~bp. Twenty-five selected ISSR primers produced 284 bands across 15 accessions, of which 270 were polymorphic with an average of 10.80 bands per primer. The PIC value ranges from 0.363 to 0.908 for RAPD primers, while 0.419 to 0.836 in case of ISSR. The size of amplified bands ranged from 2,800 to 240~bp. Similarity index values ranged from 0.16 to 0.95 (RAPD), 0.18 to 0.98 (ISSR) and 0.179 to 0.945 for pooled ISSR and RAPD markers data. Mantel test revealed the similar distribution pattern of the polymorphism between RAPD and ISSR markers and the correlation co-efficient (r) was 0.71384. The results indicated that both of the marker systems RAPD and ISSR, individually or combined can be effectively used in determination of genetic relationship among B. Monnieri accessions collected from different parts of Central India. It could be concluded that the information of genetic similarities and diversity among Brahmi accessions is necessary for their conservation and breeding programs.
  184. Darokar, Mahendra P. and Khanuja, Suman P. S. and Kumar Shasany, Ajit and Kumar, Sushil, Low Levels of Genetic Diversity Detected by RAPD Analysis in Geographically Distinct Accessions of Bacopa Monnieri, Genetic Resources and Crop Evolution, vol. 48, no. 6, pp. 555--558, December 2001. doi: 10.1023/A:1013800101604.
    Brahmi (Bacopa monnieri) is atraditional Indian medicinal plant known for its natural nootropicaction of saponins present in large amount in its shoots. Acollection of 24 B.monnieri accessions from differentagro-climatic zones of India and an introduction from Malaysiamaintained in the field genebank at CIMAP was analysed for RAPDvariation. Among the 40 random primers tested, 29 primers generatedone or more polymorphic bands. The number of polymorphic bandsgenerated was primer dependent, ranging from 2 to maximum of 8.Similarity matrices were generated from the RAPD data on the basis ofNei's estimates of similarity indices and dendrograms wereconstructed based on UPGMA clustering. All the accessions were foundto be in the range of 0.8–1.0 of similarity, which isindicative of a narrow genetic base among the various accessions witha medium level of polymorphism. It was possible to differentiateindividual accessions, showing differences in morphological andgrowth properties at DNA level. The observed low levels of geneticvariation were attributed to interplay of sexual and vegetative modesof reproduction and similarity of local environments in habitats ofB. monnieri.
  185. Kumar, M M and Gopi, R and Lakshmanan, G M A and Panneerselvam, R and Raj, E E, Study on Genetic Diversity of Bacopa Monnieri (l.) Pennell Ecotype Variants from Tamil Nadu by the Rapd Markers, Romanian Journal of Biology - Plant Biology, vol. 58, no. 1, pp. 9--17, 2013. url: https://www.researchgate.net/publication/315886465_Studies_on_genetic_diversity_of_Bacopa_monnieri_L_Pennell_ecotypes_variants_from_South_India_by_the_RAPD_markers.
    Brahmi is a small prostrate herb that grows wild in marshy and damp places near water logs throughout India. The plant has been used since ancient time in folklore and traditional medicine as nervine, cardio-tonic, and diuretic. The plants were collected from18 different geographical regions of south India. RAPD profiles in order to assess diversity and the similarity matrices were generated from the RAPD data on the basis of Jaccard similarity coefficient estimates of similarity indices and dendrogram were constructed based on UPGMA clustering. OPL-05 having the maximum range of polymorphism was noticed (83.33\%) and high level of monomorphism was noticed in the primer OPT-18. The amplification range is between 100 bp to 2900 bp and the total percentage of polymorphism is (50\%) was observed. The observed narrow genetic base in B. monnieri population in Tamil Nadu may be attributed to the vegetative propagation of the species.
  186. Karthikeyan, Alagarsamy and Madhanraj, Anandaraj and Pandian, Shunmugiah Karutha and Ramesh, Manikandan, Genetic Variation among Highly Endangered Bacopa Monnieri (L.) Pennell from Southern India as Detected Using RAPD Analysis, Genetic Resources and Crop Evolution, vol. 58, no. 5, pp. 769--782, June 2011. doi: 10.1007/s10722-011-9695-4.
    Randomly amplified polymorphic DNA (RAPD) fingerprinting approach was applied to assess genetic diversity in different accessions of rejuvenating and intellect-promoting ancient ayurvedic medicinal herb Bacopa monnieri (L.) Pennell collected from four Southern Indian states, along with in vitro micropropagated samples maintained in the laboratory for 5~years. With the 10 analyzed primers, 110 distinct bands, in the size range of 250–870~bp were observed, and among which 14 (12.72\%) were polymorphic. Among the random primers used, only OPD 02 generated highest percentage of polymorphism with 30.77. Whereas OPD 08 generated a maximum of 18 amplified bands, but with only 1\% polymorphism. Cluster analysis done on the basis of similarity co-efficient generated from RAPD profiles indicated all the accessions were divided into two sub-groups based on genetic distances under one major cluster. Major cluster is only the lone loose group of the Bm.2 collected from Kerala (KER) and in vitro micropropagated plant (IVMP) maintained in the lab. The other sub-group consisted of Bm.1 and Bm.3 collected from Tamil Nadu (TN) and Andhra Pradesh (AP) respectively, which are genetically similar and showed similarity with accessions from Karnataka Bm.4 (KK). These two sub-groups were joined together at 0.66 genetic distance level. Overall, the levels of genetic similarity within the accessions varied from 0.24 to 0.80, the matrix ranged from 0.36 to 0.80, with a mean value of 0.68 indicating genetic similarity at low level.
  187. Sharma, Abhishek and Verma, Neha and Verma, Priyanka and Verma, Ram Kishor and Mathur, Archana and Mathur, Ajay Kumar, Optimization of a Bacopa Monnieri-Based Genetic Transformation Model for Testing the Expression Efficiency of Pathway Gene Constructs of Medicinal Crops, In Vitro Cellular \& Developmental Biology - Plant, vol. 53, no. 1, pp. 22--32, February 2017. doi: 10.1007/s11627-017-9804-y.
    A fast regenerating Agrobacterium tumefaciens-mediated transformation protocol for Bacopa monnieri (L.) Wettst. was developed as a model system for heterologous expression of terpenoid indole alkaloid pathway genes from Catharanthus roseus (L.) G. Don. The direct regeneration of shoots from leaf explants co-cultured with A. tumefaciens resulted in the integration of a tryptophan decarboxylase (tdc) and strictosidine synthase (str) cassette (
  188. Croom, LaMont Alexander and Jackson, Carissa L. and Vaidya, Brajesh Nanda and Parajuli, Prahlad and Joshee, Nirmal, Thin Cell Layer (TCL) Culture System for Herbal Biomass Production and Genetic Transformation of Bacopa Monnieri L. Wettst., American Journal of Plant Sciences, vol. 7, no. 8, pp. 1232--1245, June 2016. doi: 10.4236/ajps.2016.78119.
    Bacopa monnieri (L.) Wettst. (Scrophulariaceae) is a highly sought after medicinal plant with therapeutic properties as cognition enhancer as well as for other brain and body functions. Research was conducted to optimize a thin cell layer explant based micropropagation system to assist mass propagation. Thin cell layers (TCL) derived from leaf and internode segments were used as explants. Murashige and Skoog medium was used to formulate shoot induction, elongation, and rooting media. Shoot induction media were prepared by supplementing three concentrations (0.1, 1.0, and 10.0 μM) of four cytokinins 6-benzylaminopurine, 2-isopentenyl-adenine, 6-3-Hydroxybenzylaminopurine, and thidiazuron to study adventitious shoot bud induction response. An optimum shoot bud induction response was observed on MS medium supplemented with 10.0 μM 6-benzylaminopurine for both leaf and stem transverse thin cell layer (tTCL) explants. The average number of shoot buds from leaf tTCL explants was 59, whereas, on an average, 33 shoot buds were regenerated from internode tTCL explants. Elongation of adventitious shoot buds was achieved best in a liquid medium using Liquid Lab Rocker® system. Elongated shoots recorded 100\% rooting in MS medium supplemented with 5 μM indole butyric acid. Bacopa micropropagation employing tTCL explants for initial shoot bud induction and using LLR® boxes in subsequent elongation step can achieve cost effective way to regenerate high volume of plantlets and biomass required for herbal industry. Leaf and stem tTCL explants both were suitable for Agrobacterium tumefaciens (EHA105) mediated genetic transformation. Successful transformation was scored within three days of co-cultivation with Agrobacterium suspension on the basis of Enhanced Green Fluorescent Protein (EGFP) expression as an early and non-destructible screening device. Transformation frequencies of 83\% and 76\% were accomplished for leaf and stem tTCL explants, respectively. Greenhouse grown Bacopa plants were analyzed as fresh and dry methanolic extracts for total polyphenol content (811.93 ± 7.98 and 814 ± 17.64 GAE mg g-1) and the Trolox Equivalent Antioxidant Capacity values were 1918.25 ± 173.12 and 3163.14 ± 403.25 μmol/g, respectively.
  189. Shalini, Vijayanna Tirumalapura and Neelakanta, Sajjanar Jambappa and Sriranjini, Jaideep Sitaram, Neuroprotection with Bacopa Monnieri–A Review of Experimental Evidence, Molecular Biology Reports, vol. 48, no. 3, pp. 2653--2668, March 2021. doi: 10.1007/s11033-021-06236-w.
    Brahmi or aindri is a popular herb in the vast and rich compendium of herbs of Ayurveda and is botanically identified as Bacopa monnieri Linn. (BM). It is extensively used in Ayurveda and other traditional systems of medicine in the management of neurological psychiatric disorders. BM possess active principles belonging to alkaloids, glycosides, flavonoids, saponins categories. Numerous research have been undertaken across the globe to evaluate the neuroprotective potential of this herb. This review collates and summarises current (as on May 2020) published literature on Brahmi as a neuroprotective in neurological and psychiatric disorders. English language articles from databases PubMed, Scopus and Google scholar were searched using appropriate free keywords and MeSH terms related to the topic. The review demonstrates the neuroprotective potential of the Ayurveda herb Brahmi in several disorders including Alzheimer’s disease, epilepsy, Parkinson’s disease, Huntington’s disease, cerebral ischemia and infarct and neoplasms.
  190. Jain, Pushpendra Kumar and Das, Debajyoti and Jain, Puneet and Jain, Prachi, Pharmacognostic and Pharmacological Aspect of Bacopa Monnieri: A Review, vol. 4, no. 3, pp. 5, 2016.
    It is said that the use of Bacopa monnieri (BM) for memory enhancement goes back 3000 years or more in India, when it was cited for its medicinal properties, especially the memory enhancing capacity, in the vedic texts “Athar-Ved Samhila” (3:1) of 800 BC and in Ayurveda. In the folklore of Indian medicine, several herbs have been used traditionally as brain or nerve tonics. One of the most popular of these neurotonics is BM, a well-known memory booster. Brahmi has been administered at religious institutions to help students to enhance their memory for learning ancient, religious hymns. It is also used as cardio-tonic, tranquilizer and sedative, improves the process of learning, restores memory, and enhances power of speech and imagination, diuretic and nervine tonic, antistress, for nervous and mental strain, use in insanity, epilepsy, hysteria, esthenia, nervous breakdown. It is a small, creeping succulent herb. The leaf and flower bearing stems are 10-30 cm long and arise from creeping stems that form roots at the nodes with pale blue or pinkish white flowers belonging to family Scrophulariaceae grown nearly banks of freshwater streams and ponds, paddy fields, and other damp places. The chief phytoconstituents present are brahmine, herpestine, alkaloids, and saponins.