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Increased miroestrol, deoxymiroestrol and isoflavonoid accumulation in callus and cell suspension cultures of Pueraria candollei var. mirifica

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Abstract

Miroestrol and deoxymiroestrol are highly active phytoestrogens derived from the tuberous roots of Pueraria candollei var. mirifica. To date, there have been no reports regarding the production of miroestrol and deoxymiroestrol in in vitro cell culture. In this study, callus and cell suspension cultures were established for the purpose of investigating miroestrol and deoxymiroestrol content in P. candollei var. mirifica cells. Stem-derived callus cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg l−1 thidiazuron (TDZ), 0.5 mg l−1 naphthaleneacetic acid (NAA), and 1.0 mg l−1 benzyladenine (BA) provided optimal conditions for the accumulation of deoxymiroestrol and total isoflavonoids. The calli produced 184.83 ± 20.09 μg g−1 dry weight of total chromene and 20.72 ± 2.38 mg g−1 dry weight of total isoflavonoid. This is the first report to suggest that callus culture is a suitable alternative method for producing miroestrol and deoxymiroestrol. Carbon sources were evaluated for the cell suspension cultures of P. candollei var. mirifica. Sucrose provided optimal conditions for biomass production, whereas fructose was the most suitable carbon source for deoxymiroestrol and isoflavonoid production. The information from our study can be employed for enhancing the production of miroestrol, deoxymiroestrol, and total isoflavonoids using in vitro cell culture of P. candollei var. mirifica.

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Abbreviations

BA:

6-Benzyladenine

GPBS:

0.2% Gelatin in phosphate-buffered saline

NAA:

α-Naphthaleneacetic acid

PAbs:

Polyclonal antibodies

TDZ:

Thidiazuron

TPBS:

0.05% Tween 20 in phosphate-buffered saline

Wt:

Weight

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Acknowledgments

This work was supported by a grant from the Office of the Higher Education Commission. Latiporn Udomsuk (CHE 510636) received a CHE-Ph.D. scholarship. The Thailand Research Fund (RSA5280012) is gratefully acknowledged. We also thank Dr. Chaiyo Chaichantipyuth, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand, for providing purified miroestrol and deoxymiroestrol.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Latiporn Udomsuk, Kanokwan Jarukamjorn & Waraporn Putalun

  2. Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, 40002, Thailand

    Latiporn Udomsuk, Kanokwan Jarukamjorn & Waraporn Putalun

  3. Faculty of Pharmaceutical Sciences, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand

    Thaweesak Juengwattanatrakul

Authors
  1. Latiporn Udomsuk
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  2. Thaweesak Juengwattanatrakul
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  3. Kanokwan Jarukamjorn
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  4. Waraporn Putalun
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Corresponding author

Correspondence to Waraporn Putalun.

Additional information

Communicated by J. V. Jorrin-Novo.

Electronic supplementary material

Supplementary Fig. 1. Characteristic of calli of Pueraria candollei var. mirifica by different type and ratio of growth regulators. Calli were harvested at 8 weeks of culture. a Combination of TDZ and NAA, and b combination of TDZ, NAA, and BA.

Supplementary Fig. 2. Characteristic of cell suspensions of Pueraria candollei var. mirifica in MS medium supplemented with 0.1 mg l−1 TDZ, 1.0 mg l−1 NAA, and 0.5 mg l−1 BA by different type sugar (3% W/V). Cell suspensions were harvested at 4 weeks of culture. a Dextrose, b sucrose, and c fructose.

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Udomsuk, L., Juengwattanatrakul, T., Jarukamjorn, K. et al. Increased miroestrol, deoxymiroestrol and isoflavonoid accumulation in callus and cell suspension cultures of Pueraria candollei var. mirifica . Acta Physiol Plant 34, 1093–1100 (2012). https://doi.org/10.1007/s11738-011-0906-6

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  • DOI: https://doi.org/10.1007/s11738-011-0906-6

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