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Article

Allium pallasii and A. caricifolium—Surprisingly Diverse Old Steppe Species, Showing a Clear Geographical Barrier in the Area of Lake Zaysan

by
Nikolai Friesen
1,2,*,
Lisa Grützmacher
1,
Mikhail Skaptsov
3,
Polina Vesselova
4,
Vladimir Dorofeyev
5,
Alexander N. Luferov
2,
Nazgul Turdumatova
6,
Georgii Lazkov
6,
Sergei V. Smirnov
3,
Alexander I. Shmakov
3 and
Herbert Hurka
1
1
Botanical Garden, University of Osnabrück, Albrechtstrasse 29, 49076 Osnabrück, Germany
2
Department of Pharmacy and Natural Sciences, A.P. Nelyubin Institute of Pharmacy, I.M. Sechenov First Moscow State Medical University, Ministry of Health of the Russian Federation, Izmailovsky Boulevard 8, 105043 Moscow, Russia
3
South Siberian Botanical Garden, Altai State University, Lenina Str. 61, 656049 Barnaul, Russia
4
Institute of Botany and Phytointroduction of the Committee of Forestry and Wildlife, Ministry of Ecology, Geology and Natural Resources of the Republic of Kazakhstan, 480070 Almaty, Kazakhstan
5
Komarov Botanical Institute of RAS, Professor Popov Str. 2, 197376 Saint Petersburg, Russia
6
Institute of Biology, National Academy of Sciences, Bishkek 720071, Kyrgyzstan
*
Author to whom correspondence should be addressed.
Plants 2022, 11(11), 1465; https://doi.org/10.3390/plants11111465
Submission received: 18 March 2022 / Revised: 19 May 2022 / Accepted: 19 May 2022 / Published: 30 May 2022
(This article belongs to the Section Plant Systematics, Taxonomy, Nomenclature and Classification)
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Versions Notes

Abstract

:
Polymorph Allium pallasii s.l. from monotypic A. sect. Pallasia was studied using a wide spectrum of methods and divided into two clearly morphologically, geographically, cytologically and genetically isolated species: A. pallasii s. str.—North-East Kazakhstan, Western Siberia, and the Altai Mountains; A. caricifolium—Kyrgyzstan, Northwest China, South-East Kazakhstan until Zaysan Lake in the east. Despite serious genetic differences, both species are sisters and are related to species of the A. sect. Codonoprasum (Subg. Allium). Allium caricifolium differs from A. pallasii s. str. by taller stems, dense inflorescence, and with filaments longer than perianth. The possible phylogenetic reasons for the separation of these species are discussed. A nomenclature analysis of synonyms was carried out.

1. Introduction

Allium L. (Amaryllidaceae J.St.-Hil.: Allioideae Herb.) is one of the largest monocot genera with more than 1000 species [1] naturally distributed throughout the northern hemisphere [2,3,4,5,6,7]. The main centres of biodiversity are located in arid and sub-arid regions of Southwestern and Central Asia, and in the Mediterranean region. The significantly smaller centre is in western North America [5,7,8,9]). The genus is characterized by bulbs (often formed on rhizomes) enclosed in membranous, fibrous, or reticulate tunics, free or basally connate sepals, and usually a subgynobasic style [7]. The overwhelming morphological diversity in the genus is mirrored by a complicated taxonomic structure consisting of 15 subgenera and 72 sections of three evolutionary lineages [4,7].
All subsequent phylogenetic studies [7,9,10,11,12,13,14,15] confirmed the division of Allium into three major evolutionary lineages with the monophyletic origin of all subgenera included in the first and second evolutionary lineages. The phylogenetic relationships in the youngest third lineage are less clear. According to the latest studies, many subgenera are not monophyletic in the third evolution line. This mainly affects the subgenera Cepa (Mill.) Radić, Reticulatobulbosa (Kamelin) N.Friesen, Rhizirideum G.Don ex Koch) Wendelbo, Polyprason Radić and possible Allium [5,9,12,13,16].
Allium pallasii Murrey [17], the single representative of the monotypic A. sect. Pallasia (Tsag.) F.O. Khas., R.M. Fritsch & N. Friesen (subgen. Allium), is widespread in the steppe from central Kazakhstan in the west and to the Kulunda Steppe in West Siberia, Russia in the east and from 54° N in the north in Kazakhstan to Kyrgyzstan, south-east Uzbekistan, and north-west China in the south. There are two isolated occurrences of A. pallasii in the Kurai Steppe in the Altai Mountains and the Dzungarian Gobi in East Mongolia [18,19,20]. From the beginning, we were interested in the relationship of plants from Altai with the A. pallasii plants from South Kazakhstan and Kyrgyzstan. However, the plants from Altai and northern Kazakhstan differ morphologically from plants from southern Kazakhstan and Kyrgyzstan, with smaller flowers and longer pedicels, giving the impression of fewer flowers per inflorescence [21,22,23]. The filaments are also no longer than tepals, in contrast to plants from southern Kazakhstan and Kyrgyzstan, which have long filaments. Despite such obvious morphological differences, many botanists regard A. pallasii as a highly variable species [21,22], and some [23], having discovered significant karyological and morphological differences between plants from northern Kazakhstan and Kyrgyzstan, have not dared to recognize them as distinct species. Allium pallasii is throughout the distribution area diploid, with 2 n = 16 chromosomes [23,24,25].
Preliminary ITS sequences of plants from the Kurai steppe (Altai, Russia) and Kyrgyzstan displayed surprisingly different sequences. First investigations into taxonomy showed the great complexity of the taxonomy and nomenclature of A. pallasii as shown by the seven synonyms (A. tenue G.Don, A. lepidum Ledeb., A. caricifolium Kar.et Kir.,, A. nitidulum Fisch. ex Ledeb., A. alberti Regel, A semiretschenskianum Regel, and A. saxatile Hohen ex Boiss) in Plants of the World Online [26].
To shed light on these phylogenetic and taxonomic problems, plants from the entire distribution area of A. pallasii s. l. were systematically collected from 2010 and included in the geographical, morphological, cytological, and molecular analysis.

2. Results

2.1. Morphology and Distribution

Morphological analysis of herbarium sheets and our collections of the A. pallasii accession revealed a clear dividing line between the two morphotypes in the A. pallasii s.l.: plain steppe morphotypes and mountain steppe morphotypes. To the north of Lake Balkhash and to the east of Lake Zaysan Allium pallasii s. str. is widespread in plain steppe and south of this line only the mountain morphotype (A. caricifolium) with taller stems, dense inflorescence, and filaments 1.25 times longer than tepals, is found (Figure 1). Both species have apparent morphological differences; a comparison with some morphological features is presented in Table 1 and can be seen in Figure 2, Figure 3 and Figure 4.

2.2. Phylogenetic Analyses

2.2.1. Position of A. pallasii s.l. in the Third Evolution Line

The alignments of nrITS sequences (including the 5.8S gene) with 253 accessions of Allium species, a selection of representatives from each subgenus, and sections of the third evolutionary line, including five accessions of A. pallasii s. str. and five accessions from South Kazakhstan and Kyrgyzstan (A. caricifolium), consist of 732 characters of which 517 variable characters are parsimony informative. Unweighted parsimony analysis of the 253 sequences resulted in about 17 million most parsimonious trees of 4361 steps (CI = 0.2740). The substitution model TVM + G was chosen by AIC in JModeltest-2.1.7 for the Bayesian analysis. The Parsimony and Bayesian analyses produced identical topology (See Figure S2). All A. pallasii s. l. accessions are divided into two sister groups and stand surprisingly as a sister group to the A. sect. Codonoprasum of subgenus Allium. Both species (A. pallasii and A. caricifolium) are sister groups also in the plastid tree (rpl32-trnL) with 115 accessions from most sections of the third evolutionary line and five accessions from A. pallasii s.l. Both clades of A. pallasii and A. caricifolium together are entitled as a sister clade to A. sect. Codonoprasum (See Figure S3). The generalized ITS tree with sections and subgenera names is shown in Figure 5. Some subgenera in the third evolution line after classification [7] are not monophyletic; this applies to subgenera Cepa, Reticulatobulbosa, Polyprason, Rhizirideum, and possible Allium. These results agree with previously published phylogenetic analyses [5,9,11,12,13,27,28]. The phylogenetic consequences for the non-monophyletic subgenera should be made in the future with detailed analysis, but here is the most important finding for us, that the A. sect. Pallasii is a sister group to A. sect. Codonoprasum with strong support. The matching of the two sister clades A. pallasii and A. caricifolium is only moderately supported: Bayesian posterior probabilities (PP) = 0.86 and bootstrap support (BS) = 70.

2.2.2. Phylogeny of Allium pallasii s.l.

Furthermore, we made a phylogenetic screening with 51 accessions A. pallasii s.l., carried out from the entire distribution area (31 accessions of A. pallasii and 20 accessions of A. caricifolium) with three representatives of the A. sect. Codonoprasum (A. flavum L., A. paniculatum L. [29,30] and A. praescissum Rchb. [31]) as outgroup with nuclear (ITS) fragments and two plastids (trnL-rpl32 and trnQ-rps16). ITS sequences within A. pallasii and A. caricifolium are monomorphic, with rare single-nucleotide swaps. Especially the accessions of A. pallasii s. str. have identical sequences. Only in the mountainous morphotype (A. caricifolium), are the accessions from the Alai Mountains in Kyrgyzstan grouped into a clade with relatively good support. Both species are divided into two sister groups with very high support because the sequences are very different (See Figure S3a). In the BLAST analysis, the nrITS sequences from A. pallasii s. str. were only 84.04% similar to A. caricifolium sequences from Northwest China (as A. pallasii in NCBI GenBank: GQ181077 China; KF693249 China: Xinjiang, Urumchi; KF693250 China, Xinjiang, Zhaosu), which correlates well with the group mean distance between A. pallasii and A. caricifolium ITS sequences (P = 0.188).
We obtained similar results with plastid sequences, where the polymorphism within morphotypes is significantly higher than with nrITS sequences. See the plastid tree in Figure S3b. There are only two discrepancies regarding the position of accession Am579 and Am606. In the ITS tree accession Am579 stays within A. pallasii s. str. clade and in the plastid tree clearly below the A. caricifolium clade. The situation at accession Am606 is reversed (Figure S3). This is a clear indication of the hybrid origin of these accessions. In addition, their location in the border regions between both species supports the hybridogenic origin (Figure 1). Except for these two cases, the topology of the trees is very similar, so we aligned and analysed all the sequences together (Figure 6). Both hybrid accessions are expected to stand apart in the tree, but both sister clades are clearly monophyletic with very strong support. There are a few small groupings in A. pallasii clade with weak support; only one subclade with four accessions (Am189, Am482, Am574, Am575) has strong support (PP = 0.97; BS = 95). All these accessions are from the easternmost distribution. In the BLAST analysis, the trnL-rpl32 spacer sequences from A. pallasii s. str. were only 94.19% similar to A. caricifolium sequence from Northwest China (as A. pallasii in NCBI GenBank: MN648632 complete chloroplast genome).
Within A. caricifolium clade are two subclades with good support: accessions Am580, Am1275, Am1280 and 1281 form one, and two accessions from Alai valley in Kyrgyzstan are the second well-supported clade. In the first subclade, two accessions are from Chu valley in Kyrgyzstan, one from Alay valley (Am1275), and one (Am580) is from the western part of the Zaysan lowland.

2.3. Cytology, Flowcytometry

From A. pallasii, we examined the karyotypes of four accessions (Am457, Am588, Am776, Am781). All four accessions have similar chromosome morphology. Two middle pairs of chromosomes have very small dot satellites in the shorter arm. There are only metacentric chromosomes in the karyotype of A. pallasii. Therefore, we calculated a combined idiogram of 36 metaphases (Figure 7a, Table 2. For A. caricifolium, we could study the chromosome morphology of the accession Am1246. The sixth pair of chromosomes are metacentric, and two satellite chromosomes are submetacentric. Compared to A. pallasii, the satellites are massive in A. caricifolium, between one and two µm (Figure 7b, Table 3). Overall, the chromosomes in A. caricifolium are also slightly larger. Total karyotype diploid length (TKL) in A. pallasii = 87.14 µm and in A. caricifolium = 103.19 µm. This correlates well with the estimated genome size by flow cytometry in both species: A. pallasii 2C = 14.03 pg (Am607); A. caricifolium 2C = 20.37 pg (Am708). See the histograms of relative DNA content in Figure S5.

2.4. Nomenclatural Remarks

Murray [17] described A. pallasii on the plants grown in the botanical garden of the University of Göttingen from seeds, sent by P.S. Pallas, without geographical origin. According to the description and analysis of the picture [17] (Table 1) and type material, the name A. pallasii belongs to the plain steppe morphotypes of plants. The species named A. tenue G.Don [34] is also based on the samples from the Herbarium of Pallas. The description staminibus perianthio aequalibus” clearly refers to it as a synonym of A. pallasii s. str. The species named A. lepidum for the plain steppe morphotypes was donated by Ledebour [35] that included in the protologue a short description and an illustration (Table CCCLV) strangely named A. pallasii. The other synonym name for plain steppe morphotypes in POWO [26] is A. nitidulum Ledeb.
This name is a nomen nudum cited by Ledebour [36] as “A. nitidulum Fisch. in herb. reg. berol.” as synonym of A. pallasii. That means Ledebour, in both cases (A. lepidum and A. nitidulum), did not recognize the species status for Altai plants, but ultimately included them in synonyms to A. pallasii. Only Regel [37] first validated the name “A. nitidulum” as a variety of A. pallasii. The name of A. saxatile Hohen. ex Boiss. (nom.illeg.) was also incorrectly cited as synonym to A. pallasii [26]. It is a synonym of A. kunthianum Vved. [21].
All other names regarded as synonyms of A. pallasii s. l. (i.e., A. caricifolium, A. alberti, and A. semiretschenskianum) belong to the southern mountain morphotype with priority name A. caricifolium. Allium caricifolium Kar. & Kir. is described on the plants collected in the Mountains near the Ajagus settlement. The morphological character in the description “… staminibus simplicibus, basi subuîatis, perigonium subdupio excedentibus“ unequivocally refers to the mountainous morphotype from south-east Kazakhstan and Kyrgyzstan [38], and is typified by [39] (lectotype MW0591659). Regel had correctly identified the differences between A. pallasii plants and the plants from southern Kazakhstan and described the plants from the Almaty region (formerly Vernoe) as A. semiretschenskianum [40]. The name A. caricifolium Regel had been unfortunately placed as a synonym for A. pallasii [41]. The situation with the name A. alberti Regel [41] is a bit complicated. This species was described from plants grown in the garden from bulbs collected by Albert Regel in the Chinese part of the Ili River in 1876 [42]. The lectotype of A. alberti (LE01010227, designated by [43]) shows a bulbless plant, and the morphological characters show extreme similarity with A. caricifolium and A. semiretschenskianum (Lectotype of A. semiretschenskoanum LE000518202 designated here, LE00052546). But Regel’s original description of A. alberti is slightly confusing. He gives a detailed description of the slender reticulate-fibrous outer tunics of the bulbs “Bulbi ovati tunicis exterioribus tenuibus totis reticulate-fibrosis, …” and gives A. moschatum L. and A. sindjarense Boiss. & Hausskn. ex Regel (A. sect. Scorodon) as related species. All other characteristics in the description match A. caricifolium very well. We can only guess whether, or not, this is a mix-up with another bulb that his son Albert Regel [42] collected during his trip to China. It was Vvedensky [21] who put both species A. semiretschenskianum and A. alberti as synonyms to A. pallasii, and we put these two names as synonyms to A. caricifolium.
Section Pallasia (Tzag.) F.O.Khass., R.M.Fritsch & N.Friesen—2017: 87 [44]. Khasanov F. in Sennikov (ed.) Flora of Uzbekistan. Vol 1. ≡ Section Pallasia (Tzag.) F.O.Khass., R.M.Fritsch & N.Friesen in [7], nom. invalid. ≡ Allium ser. Pallasia Tzag. Bot. Mater. Gerb. Bot. Inst. Bot. Acad. Nauk Kazakh. S.S.R. 11: 44 (1979 [45]).
Type: Allium pallasii Murray
Allium pallasii Murrey, Novi Comment. Soc. Regiae Sci. Gott. vi. (1775) 32. t. 3.—Lectotype (designated by 45: 87): Herb. Murrey. Described from plants grown in the botanical garden of the University of Göttingen from seeds, sent by P.S. Pallas. without geographical origin. (MW barcode MW0591688; image of the lectotype available at https://plant.depo.msu.ru/open/public/item/MW0591688, accessed on 1 April 2022)
= A. lepidum Ledeb. 1833. Icon. Pl. 4: 17, Table CCCLV.
Type ? Ilab. in collibus apricis et campestribus siccis ad fl. Irtysch a fortalitio Ustkamenogorsk usque ad lacum, qui Noor—Saisan vacatur. Fl. 3 Iajo. 7 J.
=A. tenue G.Don, Mem. Wern. Nat. Hist. Soc. vi. (1827): 34.
Type ? The description of this plant, and the preceding one, were taken from specimens in the Herbarium of Prof. Pallas, now in the possession of Mr. Lambert [34].
=A. pallasii var. nitidulum (Fisch.) Regel, Trudy Imp. S.-Petersburgs. Bot. Sada 10: 317 (1887).
Type: Herb. Fischer. No. 127. Collected near the Chuya River in meadows. July of the 10th day (LE!)
Description—Bulb ovoid, 12 mm diam., with outer gray, almost leathery shells. Shells with clear parallel veins. Stems 120–200 (290) mm high, covered by leaf sheaths up to 1/3–1/2 of its length. Leaves 2–3, filiform, semi-cylindrical, smooth, shorter than stem. Spathe 2 (3) times shorter than umbel, shortly pointed. Inflorescence is hemispherical or more often spherical, many-flowered, loose. Pedicels are almost equal between themselves, 2–3 (4) times longer than perianth. The tepals are pink, with a purple vein, shiny, 3 mm long, equal in length, lanceolate, and acute. The filaments of the stamens do not exceed the length of the perianth, subulate, and are slightly widened internally at the base. The style of the pistil is equal to or slightly longer than the perianth.
Distribution—Central and Northeast Kazakhstan, West Siberia (Kulunda Steppe), and West Altai. Two isolated distributions are in the Kurai Steppe (southeast Altai Mountains) and in Dzungarian Gobi (West Mongolia).
Habitat—In sandy and fescue steppes, on salt licks
Allium caricifolium Kar. et Kir., Bull. Soc. Imp. Naturalistes Moscou 14: 854 (1841).
Lectotype (designated by 39: 39). In montosis sterilibus prope Ajagus, nec non in apricis montium Aktschauly et Tarbagatei ad torrentes Dschanybek et Terekty. Leg. Karelin et Kirilov a. 1840. (MW! Barcode MW0591659!, the image of the lectotype available at https://plant.depo.msu.ru/open/public/item/MW0591659, accessed on 1 April 2022)
=A. semiretschenkianum Regel, Trudy Imp. S.-Peterburgsk. Bot. Sada v. (1877) 630.
Lectotype: Balchasch s.d., s. coll. (LE! Barcode LE000518202).
=A. albertii Regel 1878 in Acta Horti Petrop. 5: 632.
Type: Bulbe leg. A. Regel prope Suidun ad fluvium Ili. Ex horto bot. Petropolitani. 78.5. (LE! LE01010227)
Description—Bulb ovoid, 10–20 mm thick, outer shells gray, papery, without veins. Stem 20–65 cm high, 1/3 or almost up to 1/2 covered with smooth leaf sheaths. Leaves 3–4, filiform or narrowly linear, 1.5 (2.5) mm wide, shorter than the stem. Spathe 2–3 times shorter than an inflorescence, shortly pointed. Inflorescence spherical, many-flowered, dense. Pedicels are almost equal, 2–3 times longer than perianth. Tepals are pink with a purple vein, shiny, 3–4 mm long, equal, lanceolate or oblong-lanceolate, and acuminate. Filaments of stamens up to 1.5 times as long as tepals, subulate from a triangular base, inner base wider than the outer ones. The style of the pistil is slightly longer than the perianth.
Distribution–To the west of the Zaisas basin, Tarbagatai, Dzungarian Alatau and Central and Eastern Tian Shan
Habitat–On fine earth, gravelly and rocky slopes, outcrops of variegated rocks in the mountainous and subalpine belt

3. Discussion

All of our results (morphological, geographical, cytological, and molecular) quite clearly confirm the presence of two very well separated species in the formerly monotypic A. sect. Pallasia: A. pallasii s. str., and A. caricifolium. Li et al. [9] erroneously introduced several other Allium species into the section, mostly belonging to A. sect. Caerulea [46]. Complete chloroplast genome analysis of seven Chinese species (A. delicatulum, A. schoenoprasoides, A. songpanicum, A. tanguticum, A. caeruleum and A. teretifolium, including A. pallasii (A. caricifolium) from northwest China [27]), supports the isolated position of section Pallasia. The plastid genome of Chinese A. pallasii (MN648632) and nrITS sequences (GQ181077, KF693249, KF693250) belong to the A. caricifolium. It is possible that A. pallasii s. str. also occurs in the border region east of the Black Irtysh River (see Figure 1). So far, we have seen no evidence of this.
We confirmed 2 n = 16 for both morphotypes as expected from earlier studies [23] for both species and for A. caricifolium [23,24]. Vakhtina & Kudryashova [23] studied the morphology of the chromosomes of both morphotypes (A. pallasii from North East Kazakhstan and A. caricifolium from Transili Alatau) and found that both karyotypes differ in the position and the size of the satellites in the satellite chromosomes. Our data confirm these differences. Differences in plant morphology were also well recognized, but unfortunately, no consequent conclusions were made [23]. Simply A. pallasii was declared as very polymorphic.
It is also very surprising that the sequences of both species are so different (only 84% similarities in ITS sequences) and still grouped as a sister subclade. The closest relationship to A. sect. Codonoprasum cannot be explained morphologically either. Morphologically A. pallasii and A. caricifolium are more like species from the A. sect. Caerulea (e.g., with A. delicatulum, A. caesium, and others), which explains the inclusion of some species by Li et al. [9] in A. sect. Pallasia. In the nrITS and plastid trees, the species from A. sect. Caerulea are relatively distant from A. sect. Pallasia (Figure 1, Figures S1 and S2). When comparing the genetic differences between A. pallasii and A. caricifolium with other Allium species where times of evolutionary splits were estimated [16,28], we hypothesize an Oligocene split between A. sect. Codonoprasum and A. sect. Pallasii, and between A. pallasii and A. caricifolium Myocene split. These splits can be explained by the vegetation/landscape history of the Zaysan Depression.
The Oligocene in extratropical Eurasia is marked by the expansion of the Boreal vegetation zone (warm and humid) and the formation of temperate deciduous mesophyllous coniferous-broadleaved forests (Turgai Flora) [47,48]. In East Kazakhstan, the Turgai Flora became dominant during the Oligocene and the first half of the middle Miocene [49,50]. During the Miocene, large depressions in the hilly zone of the present-day Altai and northern Tien Shan were formed, and an inland lake has been proved for the Zaysan Depression [49]. It is suggested that a paleolake existed here since the Cretaceous period and that the Zaysan Basin was never dried [51].
Present-day Altai and northern Tien Shan mountains are believed to be of relatively recent origin (Neogene) and started to develop from the Miocene onwards as a direct result of the far-field effects of the Himalayan collision [52]. With the rising mountains, the relief energy increased and had consequences for the drainage pattern. It is hypothesized that the Altai-draining rivers flew southwards into the Zaysan and adjacent Junggar Basin, and the Tien Shan-draining rivers northwards also into the Junggar Basin [53] filling the Zaysan paleolake and creating paleolakes in the Junggar Basin. The filling of the paleolakes culminated in a united Zaysan-Junggar Basin Paleolake, which in the Late Pliocene-Pleistocene cut through the northern end of the Zaysan Basin triggering the birth and the formation of the course of the Irtysh River [53].
This scenario has consequences for the vegetation history in the Zaysan Depression. Forest vegetation (Turgai Flora) and paleolakes prevented the establishment of modern steppes for a long time, and it would appear that the steppe occurred only recently. Unfortunately, there are no Pliocene and younger paleo records from the Zaysan Basin itself but several studies from neighboring regions such as the area near Semei on the Irtysh River and the Kulunda Steppe point to a late Pleistocene/early Holocene steppe vegetation [54,55].
Based on the climate/landscape history outlined above, we suggest the following scenario of the evolutionary history of our vicarious species Allium pallasii and A. caricifolium: The original distribution area of the ancestral species was separated with the emergence of the Altai orogeny into two disjunct areas, leading to allopatric speciation. Allium pallasii s.str. survived in the Altai mountains (Kurai Steppe) and A. caricifolium in the Tian Shan and Tarbagatai mountains. With floods after the breaching of the dams of Chuya and Kurai lakes in Altai after the Ice Age [56,57], the seeds of A. pallasii were spread to the Kulunda Steppe and from there dispersed very quickly in the steppe of northern Kazakhstan. This could explain why the ITS sequences of all accessions of A. pallasii are identical. Allium caricifolium may have persisted in several places in the Tian Shan Mountains and spread north and east after the Ice Age, where it met with A. pallasii at Lake Zaysan. Similar splits between northern Kazakhstan, western Siberia, including the right bank of the Irtysh River up to the Altai mountains in one site, and mountainous regions in south-eastern Kazakhstan, west of Zaysan Lake in the second, have recently been discovered and molecularly confirmed in other taxa of the genus Allium: sect. Oreiprason [58]; Allium tulipifolium Ledeb. and A. robustum Kar. et Kir. [28]; A. obliquum L. [59], and also in other plant groups: genera Krascheninnikovia (family Amaranthaceae) [60] and Goniolimon (family Plantaginaceae) [61]. All this confirms the complex phylogenetic history of the steppe flora [62,63].

4. Materials and Methods

4.1. Morphological and Distribution Analyses

We compiled distribution maps from literature and online databases and analysed herbarium collections, including field collections. A total of 20 individuals of A. caricifolium from 4 Herbarium sheets were analyzed for the morphological analysis [Am705, Am715, Am1192 (see the origin in Appendix A), Am1281 OSBU-24372 (47°33′27″ N, 80°37′21″ E)]. A total of 22 individuals of A. pallasii were analyzed from 5 Herbarium sheets [Am482, Am780, Am1285 (see the origin in Appendix A), and OSBU 24849 (48°12′58″ N, 69°13′16″ E), OSBU 25938, (51°27′50″ N, 74°19′7″ E) The average with the associated standard deviation was calculated from the measurements. The measurements on the plants were made using a ruler and a magnifying glass. The data were analysed in the SPSS program (Version 28 https://www.ibm.com/products/spss-statistics accessed on 22 April 2022). A boxplot and PCA analysis were executed with this program. The PCA is based on a correlation matrix of characters (Table 1) using the Pearson correlation coefficient. In addition, a Kolmogorov–Smirnov test for normal distribution was carried out beforehand.
Published data were critically evaluated by reference to herbarium material deposited in ALTB, AA, BRNO, FRU, HAL, GAT, LE, M, MHA, TK, MW, NS, NSK, OSBU, TASH, XJA, and W [64,65]. Herbarium acronyms are according to the Index Herbariorum [66].

4.2. Taxon Sampling

Bulbs and leaf samples of more than 50 accessions of A. pallasii s. l. for DNA isolation were collected in the course of several collecting trips in Russia (Altai), Mongolia, and Kazakhstan from 2010 and growing in the Botanical Gardens in Osnabrück (Germany) and Barnaul (Russia). Some accessions of DNA were isolated from Herbarium sheets.
Newly sequenced accessions are marked with Am number in the trees, and their origin is shown in Appendix A. To determine the position of the A. pallasii in the genus, we took the available nuclear ITS sequences and rpl32-trnL (UAG) plastid fragment of accessions of the species with representatives from all sections of the third evolution line while some accessions from the first and second evolution lines were selected as the outgroup [7]. Sequences from NCBI GenBank (https://www.ncbi.nlm.nih.gov/nucleotide/ accessed on 3 December 2021) are marked with GenBank accession numbers on the trees.

4.3. DNA Extraction, Amplification and Sequencing

Total genomic DNA was isolated from leaves in silica gel using the InnuPREPP Plant DNA Kit (Analytic Jena AG) according to the manufacturer’s instructions and used directly in PCR amplification. The complete nuclear ribosomal ITS region (ITS1, 5.8S and ITS2) was amplified using the primers ITS-A [67] and ITS-4 [68]. The PCR conditions for ITS followed ref. [7]. PCR conditions and primers for the chloroplast regions trnL-rpl32 and trnQ-rps16 were described in [69]. PCR products were sent to Microsynth SeqLab (Balgach, Switzerland for sequencing. The sequences from all the individuals were manually edited in Chromas Lite 2.1 (Technelysium Pty Ltd.South Brisbane, Australia) and aligned with ClustalX [70], the alignment was manually corrected using MEGA 7 [71].

4.4. Phylogenetic Analyses

Both data sets (nrITS and the cpDNA trnL-rpl32 markers) for identifying the position of A. sect Pallasia in the third evolution line and to find the closest relatives of A. pallasii were analysed separately through Fitch parsimony with the heuristic search option in PAUP version 4.0 b10 [72]) with MULTREES, TBR branch swapping and 100 replicates of random addition sequence. Gaps were treated as missing data. The consistency index (CI) [73] was calculated to estimate the amount of homoplasy in the character set. The most parsimonious trees returned by the analysis were summarized in one consensus tree using the strict consensus method. Bootstrap analyses (BS) using 1000 pseudoreplicates were performed to assess the support of the clades [74]. Bayesian phylogenetic analyses were also performed using MrBayes 3.1.23 [75]. The sequence evolution model was chosen following the Akaike Information Criterion (AIC) obtained from jModelTest2 [76]. Two independent analyses with four Markov chains were run for 10 million generations, sampling trees every 100 generations. The first 25% of trees were discarded as burn-in. The remaining 150,000 trees were combined into a single data set, and a majority-rule consensus tree was obtained along with posterior probabilities (PP). To determine molecular variability throughout the range, more than 50 accessions of A. pallasii s.l. and three species from the A. sect. Codonoprasum as outgroup, nrITS, and two noncoding regions plastid DNA (trnL-rpl32, trnQ-rps16) were sequenced and analysed as above. The group mean distance (P) was estimated with MEGA7.

4.5. Cytology, Flowcytometry

Bulbs were planted in pots, and growing roots were used for the karyotype analysis. Root tips were excised from the bulbs and kept overnight in distilled water on ice. They were then transferred to room temperature for 20 min and pre-treated for 3 h at room temperature in an aqueous solution of 0.1% colchicine. Roots were then fixed in a freshly prepared mixture of 96% ethanol and glacial acetic acid (3:1 v/v). Root tips were stained using hematoxylin according to the protocol reported by Smirnov [77]. Well-spread metaphase plates were electronically documented (digitally photographed), and finally, the chromosomes of the best plates were measured and pairwise arranged using the KaryoType software [78]. For A. caricifolium, 5 metaphase plates from one individual were evaluated (Am1246, Appendix A), for A. pallasii, 4 individuals were used (Am776, Am781, Am588, Am457, Appendix A), which provided 3–12 usable metaphase plates. The measurements from all metaphase plates were combined here, a total of 36 metaphase plates. Because the idiograms automatically assembled by the software were not satisfactory, we manually ordered the chromosome pairs according to their length and shape. The idiograms were designed using the bar graph function implemented in MS Excel®. The terminology of [32,33] was applied.
Flow cytometry was used for the determination of DNA amount. Fresh leaf material was harvested, and ca. 0.5 cm2 leaf material was chopped with a sharp razor blade in a Tris MgCl2 buffer supplemented with propidium iodide (50 μg/mL), RNase (10 μg/mL), and 2-mercaptoethanol (0.2%) [79]. The samples were filtered through a 50-μm nylon membrane into a sample tube. Subsequent flow cytometry was performed on a Partec CyFlow PA (Partec, Münster, Germany). As an internal standard Pisum sativum ‘Ctirad’, 2 C = 9.09 pg was used [80]. Histograms were analysed using the Flowing Software 2.5.1. (Turku Bioscience Centre, Turku, Finland).

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/plants11111465/s1, Figure S1. Phylogenetic tree of third evolutionary lineage of the genus Allium, based on ITS sequences from NCBI GenBank. Figure S2. Phylogenetic tree of third evolutionary lineage of the genus Allium, based on CP DNA sequences (trnL-rpl32) from NCBI GenBank. Figure S3. A—Phylogenetic tree of A. sect. Pallasia accessions, based on ITS sequences; B—Phylogenetic tree of A. sect. Pallasia accessions, based on two combined fragments of plastid DNA (trnL-rpl32, trnQ-rps16). Figure S4. Histograms of relative DNA content were obtained after analysis of nuclei isolated from young leaf tissues of A. pallasii, accession Am607 (A) and A. caricifolium, accession Am708 (B).

Author Contributions

Conceptualization, N.F., A.I.S., S.V.S., P.V., G.L., H.H. and V.D.; methodology, N.F., L.G., V.D., A.I.S., M.S. and A.I.S.; formal analysis, N.F., L.G., V.D. and N.T.; investigation, N.F., L.G., G.L., N.T., V.D., S.V.S. and A.I.S.; data curation, N.F., G.L., A.N.L. and N.F.; writing—original draft preparation, N.F., L.G., M.S. and H.H.; writing—review and editing all authors. All authors have read and agreed to the published version of the manuscript.

Funding

We greatly appreciate the financial support by the German Research Foundation (DFG, FR 1431/8-1 to N.F.), Russian Science Foundation (RSF 19-14-00071), Agreement with the Ministry of Education and Science of the Russian Federation No. 075-15-2021-1056; National Center of Science and Technology Evaluation JSC Ministry of Education and Science of the Republic of Kazakhstan, Kazakhstan state target scientific and technical program BR10264557 “Cadastral assessment of the current ecological state of flora and plant resources of the Almaty region as a scientific basis for effective management of resource potential”.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Relevant data applicable to this research are within the paper.

Acknowledgments

We thank the curators and staff members of the following herbaria: AA, ALTB, BRNO, HAL, GAT, FRU, LE, M, MHA, TK, MW, NS, NSK, OSBU, XJA, and W for their valuable help. Special thanks go to Alexander Naumenko (Nova Zahrada s.r.o., Czech Republic) for bulbs of A. caricifolium from Kyrgyzstan (Am1246). We would like to thank the editor and three anonymous reviewers whose input improved the manuscript.

Conflicts of Interest

The authors declare no conflict of interest.

Appendix A

Table
Table A1. Origin, source, and GenBank accession numbers of Allium sequences used for phylogenetic analyses.
Table A1. Origin, source, and GenBank accession numbers of Allium sequences used for phylogenetic analyses.
AccessionArt NameCoordinatesCountryVoucherrITStrnQ-rps16rpl32trnL
Am189Allium pallasii50°14′15″ N 87°57′34″ ERUOSBU: 18243OM891893
Am457Allium pallasii50°14′14″ N 87°48′40″ ERUOSBU: 22275OM891894OM983335OM983382
Am482Allium pallasii50°14′38″ N 87°53′48″ ERUOSBU: 23174OM891895
Am574Allium pallasii51°2′49″ N 81°2′16″ EKZNS: 0012843OM891896
Am575Allium pallasii53°25′0″ N 77°57′0″ ERUNS: 0012841OM891897OM983336OM983383
Am576Allium pallasii45°58′30″ N 92°14′59″ EMNALTB: Smirnov, 19 June 1999OM891898OM983337OM983384
Am577Allium pallasii47°55′60″ N 85°7′60″ EKZALTB: Smirnov, Antonjuk, 29 May 2000OM891899OM983338OM983385
Am579Allium pallasii x A. caricifolium49°45′59″ N 82°38′9″ EKZALTB: Smirnov, Antonyuk, 20 May 2000OM891900OM983339OM983386
Am588Allium pallasii52°4′48″ N 79°42′35″ ERUALTB: Smirnov, 22 July 13OM891901OM983340OM983387
Am607Allium pallasii50°33′40″ N 79°35′14″ EKZOSBU: 23346OM891902OM983341OM983388
Am609Allium pallasii49°54′58″ N 82°40′21″ EKZTK: Prokopjev, 13 June 1982OM891903OM983342OM983389
Am610Allium pallasii47°28′57″ N 85°14′25″ EKZTK: Schischkin, 3 June 1914OM891904
Am611Allium pallasii50°38′47″ N 79°16′5″ EKZTK: Prokopjev et al., 1 June 1975OM891905
Am633Allium pallasii53°19′0″ N 75°34′60″ EKZNS: 0015875OM891906OM983343OM983390
Am634Allium pallasii51°12′0″ N 78°10′60″ EKZNS: 0015873OM891907OM983344OM983391
Am635Allium pallasii52°29′55″ N 73°5′38″ EKZNS: 0015872OM891908OM983345OM983392
Am713Allium pallasii48°9′52″ N 82°54′23″ EKZOSBU: 24384OM891909OM983346OM983393
Am775Allium pallasii48°40′45″ N 71°29′20″ EKZOSBU: 24800OM891910OM983348OM983394
Am776Allium pallasii48°5′47″ N 69°57′58″ EKZOSBU: 24841OM891911OM983349OM983395
Am777Allium pallasii48°6′4″ N 67°34′3″ EKZOSBU: 24890OM891912OM983347OM983396
Am778Allium pallasii49°6′41″ N 66°59′48″ EKZOSBU: 24875OM891913OM983350OM983397
Am779Allium pallasii48°6′4″ N 67°34′3″ EKZOSBU: 24898OM891914OM983351OM983398
Am780Allium pallasii51°53′29″ N 77°3′51″ EKZOSBU: 25000OM891915OM983352OM983399
Am781Allium pallasii48°33′43″ N 70°54′16″ EKZOSBU: 24835OM891916OM983353OM983400
Am782Allium pallasii49°14′1″ N 73°3′58″ EKZOSBU: 24770OM891917OM983354OM983401
Am786Allium pallasii50°11′54″ N 87°56′31″ ERUOSBU: 23556OM891918OM983355OM983402
Am823Allium pallasii47°48′46″ N 85°27′40″ EKZALTB: Starikov. 21 May 1990OM891919OM983356OM983403
Am1282Allium pallasii51°20′11″ N 70°5′25″ EKZOSBU: 24956OM891920OM983357OM983404
Am1283Allium pallasii49°6′41″ N 66°59′48″ EKZOSBU: 24765OM891921OM983358OM983405
Am1284Allium pallasii50°15′49″ N 66°59′32″ EKZOSBU:24931OM891922OM983359OM983406
Am1285Allium pallasii51°41′58″ N 73°25′27″ EKZOSBU: 28157OM891923OM983360OM983407
Am456Allium caricifolium47°52′6″ N 81°28′34″ EKZALTB: Kechaikin, 2 June 12OM891924OM983361OM983408
Am491Allium caricifolium47°55′60″ N 85°7′60″ EKZOSBU: 22298OM891925OM983362OM983409
Am492Allium caricifolium47°25′53″ N 84°39′11″ EKZOSBU: 22290OM891926 OM983410
Am493Allium caricifolium40°31′51″ N 72°47′11″ EKGGAT: 0017895OM891927
Am494Allium caricifolium40°16′23″ N 72°37′30″ EKGGAT: 0017900OM891928
Am553Allium caricifolium45°50′5″ N 81°19′56″ EKZTK: Goloskokov, 18 June 1959OM891929OM983363OM983411
Am578Allium caricifolium49°45′59″ N 82°32′9″ E ALTB: Kechaikin, Tjutjunic, 1 June 2012 OM983364
Am580Allium caricifolium49°15′0″ N 81°45′0″ EKZALTB: Smirnov, Antonjuk, 18 May 2000OM891930OM983365OM983412
Am606Allium caricifolium x A.pallasii46°59′40″ N 73°26′60″ EKZTK: Pavlov, 28 May 1951OM891931OM983366OM983413
Am608Allium caricifolium43°51′55″ N 74°8′19″ EKZTK: Pavlov, 11 May 1951OM891932OM983367OM983414
Am686Allium caricifolium46°55′29″ N 80°36′52″ EKZOSBU: 24335OM891933OM983368OM983415
Am692Allium caricifolium45°43′55″ N 80°31′33″ EKZOSBU: 24340OM891934OM983369OM983416
Am705Allium caricifolium45°55′58″ N 80°36′31″ EKZOSBU: 24368OM891935OM983370OM983417
Am708Allium caricifolium47°14′20″ N 80°43′6″ EKZOSBU: 24370OM891936OM983371OM983418
Am718Allium caricifolium49°51′16″ N 82°25′26″ EKZOSBU: 24390OM891937OM983372OM983419
Am1192Allium caricifolium45°39′37″ N 80°34′56″ EKZOSBU: 24354OM891938OM983373OM983420
Am1246Allium caricifolium43°0′56″ N 74°42′19″ EKGOSN:2021-0746-WOM891939OM983374OM983421
Am1274Allium caricifolium42°36′6″ N 75°53′39″ EKGGAT: 3060090OM891940OM983375OM983422
Am1275Allium caricifolium40°8′29″ N 72°21′47″ EKGGAT: 2543277OM891941OM983376OM983423
Am1280Allium caricifolium42°47′0″ N 74°32′53″ EKGFRU: Usulbaev A.K., 20 May 2019OM891942OM983377OM983424
Am1281Allium caricifolium47°34′42″ N 80°37′47″ EKZOSBU: 24328OM891943OM983378OM983425
Am826Allium caesium48°3′22″ N 68°28′33″ EKZOSBU: 24961OM891944 OM983426
Am683Allium caeruleum47°24′53″ N 80°34′56″ EKZOSBU: 24332OM891945
Am473Allium delicatulum47°52′59″ N 81°28′34″ EKZALTB: Kechaikin, 2 June 2012OM891946
Am573Allium delicatulum51°44′13″ N 94°28′26″ ERUNS: 0014638OM891947
Am712Allium delicatulum47°55′39″ N 82°4′10″ EKZOSBU: 24380OM891948 OM983427
Am749Allium zaissanicum48°49′11″ N 83°46′12″ EKZOSBU: 23940OM891949
Am1243Allium moschatum46°47′22″ N 17°18′24″ EHUNOSN: 2019-0872-WOM891950 OM983429
Am1245Allium flavum48°54′14″ N 21°57′56″ ESVKOSN:2004-0826-WOM891951OM983379OM983428
Am1239Allium vineale48°17′16″ N 16°50′36″ EAOSBU: 17147 OM891952
Am1244Allium paniculatum49°55′26″ N 42°25′22″ ERUOSN: 2018-1203-WOM891953OM983380OM983430
Am881Allium praescisum52°29′48″ N 61°58′0″ EKZOSBU: 25829OM891954OM983381OM983431
Am630Allium amphibolum49°17′14″ N 87°43′4″ ERUOSBU: 23610 OM983432
Am875Allium lineare49°6′56″ N 72°40′59″ EKZOSBU: 24823 OM983433
Am848Allium strictum49°55′58″ N 14°8′31″ ECZFRT 2015/62 OM983434
Am1286Allium scabriscapum36°2′12″ N 51°12′10″ EIRGAT 19116 OM983435
Am1240Allium kunthianum42°39′52″ N 44°37′0″ EGEOOSN: 2020-0819-W OM983436
Am1241Allium oleraceum52°21′22″ N 14°10′12″ EDEOSN: 2016-0568-W OM983437

References

  1. Govaerts, R.; Kington, S.; Friesen, N.; Fritsch, R.; Snijman, D.A.; Marcucci, R.; Silverstone-Sopkin, P.A.; Brullo, S. 2005–2020. World Checklist of Amaryllidaceae. Available online: http://apps.kew.org/wcsp/ (accessed on 20 June 2020).
  2. Stearn, W.T. European species of Allium and allied genera of Alliaceae: A synonymy enumeration. Ann. Mus. Goulandris 1978, 4, 83–198. [Google Scholar]
  3. Stearn, W.T. How many species of Allium are known? Kew Mag. 1992, 9, 180–182. [Google Scholar] [CrossRef]
  4. Fritsch, R.M. and Friesen, N. Evolution, domestication, and taxonomy. In Allium Crop Science: Recent Advances; Rabinowitch, H.D., Currah, L., Eds.; CABI Publishing: Wallingford, UK, 2002; pp. 5–30. [Google Scholar]
  5. Chase, M.W.; Christenhusz, M.J.M.; Fay, M.F.; Byng, J.W.; Judd, W.S.; Soltis, D.E.; Mabberley, D.J.; Sennikov, A.N.; Soltis, P.S.; Stevens, P.F. An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG IV. Bot. J. Linn. Soc. 2016, 181, 1–20. [Google Scholar] [CrossRef] [Green Version]
  6. Friesen, N. Chapter 1. Genus Allium: Evolution, Classification, and Domestication. In Edible Alliums: Modern Biology, Production and Uses; Rabinowitch, H.D., Brian, T., Eds.; CABI Publishing: Wallingford, UK, 2022; in press. [Google Scholar]
  7. Friesen, N.; Fritsch, R.M.; Blattner, F.R. Phylogeny and new intrageneric classification of Allium L. (Alliaceae) based on nuclear rDNA ITS sequences. Aliso 2006, 22, 372–395. [Google Scholar] [CrossRef]
  8. Nguyen, N.H.; Driscoll, H.E.; Specht, C.D. A molecular phylogeny of the wild onions (Allium; Alliaceae) with a focus on the western North American center of diversity. Mol. Phylogenet. Evol. 2008, 47, 1157–1172. [Google Scholar] [CrossRef]
  9. Li, Q.Q.; Zhou, S.D.; He, X.J.; Yu, Y.; Zhang, Y.C.; Wei, X.Q. Phylogeny and biogeography of Allium (Amaryllidaceae: Allieae) based on nuclear ribosomal internal transcribed spacer and chloroplast rps16 sequences, focusing on the inclusion of species endemic to China. Ann. Bot. 2010, 106, 709–733. [Google Scholar] [CrossRef]
  10. Wheeler, E.J.; Mashayekhi, S.; McNeal, D.W.; Columbus, J.T.; Pires, J.C. Molecular systematics of Allium subgenus Amerallium (Amaryllidaceae) in North America. Am. J. Bot. 2013, 100, 701–711. [Google Scholar] [CrossRef]
  11. Hauenschild, F.; Favre, A.; Schnitzler, J.; Michalak, I.; Freiberg, M.; Muellner-Riehl, A.N. Spatio–temporal evolution of Allium L. in the Qinghai–Tibet–Plateau region: Immigration and in situ radiation. Plant Divers. 2017, 39, 167–179. [Google Scholar] [CrossRef] [PubMed]
  12. Xie, D.F.; Yu, H.X.; Price, M.; Xie, C.; Deng, Y.-Q.; Chen, J.P.; Yu, Y.; Zhou, S.D.; He, X.J. Phylogeny of Chinese Allium species in Section Daghestanica and adaptive evolution of Allium (Amaryllidaceae, Allioideae) species revealed by the chloroplast complete genome. Front. Plant Sci. 2019, 10, 460. [Google Scholar] [CrossRef] [Green Version]
  13. Xie, D.F.; Tan, J.B.; Yu, Y.; Gui, L.J.; Su, D.M.; Zhou, S.D.; He, X.J. Insights into phylogeny, age and evolution of Allium (Amaryllidaceae) based on the whole plastome sequences. Ann. Bot. 2020, 125, 1039–1055. [Google Scholar] [CrossRef]
  14. Costa, L.; Jimenez, H.; Carvalho, R.; Carvalho-Sobrinho, J.; Escobar, I.; Souza, G. Divide to Conquer: Evolutionary History of Allioideae Tribes (Amaryllidaceae) Is Linked to Distinct Trends of Karyotype Evolution. Front. Plant Sci. 2020, 11, 320. [Google Scholar] [CrossRef]
  15. Li, Q.Q.; Zhou, S.D.; Huang, D.Q.; He, X.J.; Wei, X.Q. Molecular phylogeny, divergence time estimates and historical biogeography within one of the world’s largest monocot genera. AoB Plants 2016, 8, plw041. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Friesen, N.; Smirnov, S.; Herden, T.; Oyuntsetseg, B.; Shmakov, A.; Hurka, H. Allium species of section Rhizomatosa, early members of the Central Asian steppe vegetation. Flora 2020, 263, 151536. [Google Scholar] [CrossRef]
  17. Murray, A. Observationes botanica. In Novi Commentarii Societatis Regiae Scientiarum Gottingensis; Joann Christian Dieterich: Göttingen, Germany, 1775; Volume 6, pp. 23–40. (In Latin) [Google Scholar]
  18. Friesen, N. Rody Allium i Calloscordum Herbert. In Flora Sibiri. Tom 4 Araceae—Orchidaceae; Malyshev, L.I., Peshkova, G.A., Eds.; Nauka—Sibirskoe Otdelenie: Novosibirsk, Russia, 1987; pp. 55–97, 177–195. (In Russian) [Google Scholar]
  19. Friesen, N. The Genus Allium L. in the Flora of Mongolia. Feddes Repert. 1995, 106, 59–81. [Google Scholar] [CrossRef]
  20. Baasanmunkh, S.; Oyuntsetseg, B.; Oyundary, C.; Oyundelger, K.; Urgamal, M.; Darikhand, D.; Soninkhishig, N.; Kechaykin, A.A.; Shmakov, A.I.; Erst, A.S.; et al. The vascular plant diversity of Dzungarian Gobi in western Mongolia, with an annotated checklist. Phytotaxa 2021, 501, 1–55. [Google Scholar] [CrossRef]
  21. Vvedensky, A.I. Luk—Allium L. In Flora SSSR. Tom 4; Komarov, V.L., Ed.; Izd. Akad. Nauk SSSR: Leningrad, Russia, 1935; pp. 112–280. (In Russian) [Google Scholar]
  22. Pavlov, N.V.; Polyakov, P.P. Genus Allium L. In Flora Kazakhstana; Pavlov, N.V., Ed.; Publishing House of the Academy of Sciences of the KazSSR: Almaty, Kazakhstan, 1958; Volume 2, pp. 134–193. (In Russian) [Google Scholar]
  23. Vakhtina, L.I.; Kudryashova, G.L. Cytotaxonomic studies of some Allium species in Kazakhstan. Bot. Zhurn. 1977, 62, 539–547. [Google Scholar]
  24. Zakirova, R.O.; Nafanailova, I.I. Chromosome numbers in some species of the Kazakhstan flora. Bot. Zhurn. 1988, 73, 1493–1494. [Google Scholar]
  25. He, X.J.; Xu, J.M. Karyotypes of 4 species in Allium sect. Haplostemon Boiss. from China. Acta Phytotax. Sin. 2001, 39, 423–432. [Google Scholar]
  26. POWO. Available online: https://powo.science.kew.org/results?q=Allium%20pallasii (accessed on 1 April 2022).
  27. Xie, F.M.; Xie, D.F.; Xie, C.; Yu, Y.; Zhou, S.D.; He, X.J. Adaptation Evolution and Phylogenetic Analyses of Species in Chinese Allium Section Pallasia and Related Species Based on Complete Chloroplast Genome Sequences. BioMed Res. Int. 2020, 2020, 1–13. [Google Scholar] [CrossRef]
  28. Friesen, N.; Smirnov, S.V.; Leweke, M.; Seregin, A.P.; Fritsch, R.M. Taxonomy and Phylogeny of Allium section Decipientia (Amaryllidaceae): Morphological characters do not reflect the evolutionary history verified by molecular markers. Bot. J. Linn. Soc. 2021, 197, 190–228. [Google Scholar] [CrossRef]
  29. Linnaeus, C. Species Plantarum; Laurentii Salvii: Stockholm, Sweden, 1753; Volume 1. (In Latin) [Google Scholar]
  30. Linnaeus, C. Systema Naturae. Editio Decima, Reformata; Laurentius Salvius: Stockholm, Sweden, 1759; Volume 2. (In Latin) [Google Scholar]
  31. Reichenbach, H.G.L. Iconographia Botanica Seu Plantae Criticae; Apud Fridericum Hofmeister: Leipzig, Germany, 1827; Volume 5. (In Latin) [Google Scholar]
  32. Levan, A.; Fredga, K.; Sandberg, A.A. Nomenclature for centromeric position on chromosomes. Hereditas 1964, 52, 201–220. [Google Scholar] [CrossRef]
  33. Tzanoudakis, D. Karyotypes of ten taxa of Allium section Scorodon from Greece. Caryologia 1983, 36, 259–284. [Google Scholar] [CrossRef] [Green Version]
  34. Don, G. A monograph of the genus Allium. Mem. Wernerian Nat. Hist. Soc. 1832, 6, 1–102. [Google Scholar]
  35. Ledebour, C.F. Icones Plantarum Novarum Vel Imperfecte Cognitarum Floram Rossicam, Imprimis Altaicam Illustrantes, Centuria II; I. Deubner: Riga, Latvia, 1833; p. 16. (In Latin) [Google Scholar]
  36. Ledebour, C.F. Flora Rossica Sive, Enumeratio Plantarum in Totius Imperii Rossici Provinciis Europaeis, Asiaticis et Americanis Hucusque Observatarum; Sumptibus Librariae E. Schweizerbart: Stuttgart, Germany, 1853; Volume 4. (In Latin) [Google Scholar]
  37. Regel, E. Allii Species Asiae Centralis in Asia Media a Turcomania Desertisque Araliensibus et Caspicis Usque ad Mongoliam Crescents. Acta Horti Petropolitani 1887, 10, 278–362. (In Latin) [Google Scholar]
  38. Karelin, G.S.; Kirilow, I.P. Enumeratio Plantarum in Desertis Songoriae Orientalis et in Jugo Summarum Alpium Alatau Anno 1841 Collectarum. Bull. Soc. Imp. Nat. Moscou 1842, 15, 503–542. (In Latin) [Google Scholar]
  39. Gubanov, I.A. Catalogue of Authentic Specimens of Vascular Plants of the Moscow State University Herbarium (MW); Gerbarii im. DP Syreishchikov: Moscow, Russia, 2002; p. 213. [Google Scholar]
  40. Regel, E. Descriptiones Plantarum Novarum Et Minus Cognitarum, Fasciculus VI. Acta Horti Petropolitani 1878, 5, 575–646. (In Latin) [Google Scholar]
  41. Regel, E. Alliorum Adhuc Cognitorum Monographia. Acta Horti Petropolitani 1875, 3, 1–266. (In Latin) [Google Scholar]
  42. Regel, A. Reisen in Central Asien, 1876–1879. In Dr. A. Petermann’s Mittheilungen aus Justus Perthes’ Geographischer Anstalt; Justus Perthes: Gotha, Germany, 1879. (In German) [Google Scholar]
  43. Egorova, T.V. Genus Allium. In Plantae Asian Centralis; Grubov, V.I., Egorova, T.V., Eds.; Nauka: Saint Petersburg, Russia, 1977; Volume 7, pp. 17–67. [Google Scholar]
  44. Khasanov, F. Amaryllidaceae in Flora of Uzbekistan; Sennikov, A.N., Ed.; Navro’z Publishers: Tashkent, Uzbekistan, 2017; Volume 1, p. 173. [Google Scholar]
  45. Zagolova, V.G. To the taxonomy of the section Rhizirideum of the genus Allium in Kazakhstan. Bot. Mater. Gerbarija Inst. Bot. Akad. Nauk. KazSSR 1974, 8, 16–19. (In Russian) [Google Scholar]
  46. Khasanov, F.O.; Karimov, F.; Tirkasheva, B. Taxonomic revision and Lectotypification of Allium sect. Coerulea (Omelch.) F.O.Khass. Stapfia 2013, 99, 208–234. [Google Scholar]
  47. Akhmetiev, M.A.; Zaporozhets, N.I. Paleogene events in central Eurasia: Their role in the flora and vegetation cover evolution, migration of phytochore boundaries, and climate changes. Stratigr. Geol. Correl. 2014, 22, 312–335. [Google Scholar] [CrossRef]
  48. Popova, S.; Utescher, T.; Gromyko, D.V.; Bruch, A.A.; Henrot, A.-J.; Mosbrugger, V. Cenozoic vegetation gradients in the mid- and higher latitudes of Central Eurasia and climatic implications. Palaeogeogr. Palaeoclimatol. Palaeoecol. 2017, 467, 69–82. [Google Scholar] [CrossRef]
  49. Akhmetyev, M.A.; Dodonov, A.E.; Sotnikova, M.V.; Spasskaya, I.I.; Kremenetzky, K.V.; Klimanov, V.A. Kazakhstan and central Asia—Plains and foothills. In Climate and Environment Changes during the Last 65 Million Years; Cenozoic: From Paleocene to Holocene; Velichko, A.A., Nechaev, V.P., Eds.; GEOS: Moscow, Russia, 1999; pp. 165–191. (In Russian) [Google Scholar]
  50. Popova, S.; Utescher, T.; Averyanova, A.; Tarasevich, V.; Tropina, P.; Xing, Y. Early Miocene flora of central Kazakhstan (Turgai Plateau) and its paleoenvironmental implications. Plant Divers. 2019, 41, 183–197. [Google Scholar]
  51. Dorfman, B.F. Zaysan—the Only Surviving Cretaceous Lake—May be Lost. Procedia Environ. Sci. 2011, 10, 1376–1382. [Google Scholar] [CrossRef] [Green Version]
  52. Yuan, W.; Carter, A.; Dong, J.; Bao, Z.; An, Y.; Guo, Z. Mesozoic–Tertiary exhumation history of the Altai Mountains, northern Xinjiang, China: New constraints from apatite fission-track data. Tectonophysics 2006, 412, 183–193. [Google Scholar] [CrossRef] [Green Version]
  53. Ma, Y.; Zheng, D.; Zhang, H.; Pang, J.; Wang, W.; Wang, Y.; Wu, Y.; He, H.; Stuart, F.M.; Xu, S. Plio-Pleistocene establishment of Irtysh River in Junggar, northwest China: Implications for Siberian-Arctic river system evolution and resulting climate impact. Geophys. Res. Lett. 2021, 48, e2021GL093217. [Google Scholar] [CrossRef]
  54. Kremenetsky, K.V.; Tarasov, P.E.; Cherkinsky, A.E. Holocene history of the Kazakhstan “island” pine forests. Bot. Zhurn. 1994, 94, 13–29. (In Russian) [Google Scholar]
  55. Rudaya, N.; Krivonogov, S.; Slowinski, M.; Cao, X.; Zhilich, S. Postglacial history of the Steppe Altai: Climate, fire and plant diversity. Quat. Sci. Rev. 2020, 249, 106616. [Google Scholar]
  56. Butvilovsky, V.V. Paleogeography of the Last Glaciation and the Holocene of Altai: An Event-Catastrophic Model/v.v. Butvilovsky; Publishing House Tomsk University: Tomsk, Russia, 1993; p. 252. (In Russian) [Google Scholar]
  57. Rudoy, A.N. Glacier-Dammed Lakes and geological work of glacial super floods in the Late Pleistocene, Southern Siberia, Altai Mountains. Quat. Int. 2002, 87, 119–140. [Google Scholar] [CrossRef]
  58. Seregin, A.; Anackov, G.; Friesen, N. Molecular and morphological revision of the Allium saxatile group (Amaryllidaceae): Geographical isolation as the driving force of underestimated speciation. Bot. J. Linn. Soc. 2015, 178, 67–101. [Google Scholar] [CrossRef] [Green Version]
  59. Friesen, N.; Vesselova, P.; Osmonaly, B.; Sitpaeva, G.; Luferov, A.; Shmakov, A. Allium toksanbaicum (Amaryllidaceae), a new species from Southeast Kazachstan. Phytotaxa 2021, 494, 251–267. [Google Scholar] [CrossRef]
  60. Seidl, A.; Tremetsberger, K.; Planzelt, S.; Blattner, F.R.; Neuffer, B.; Friesen, N.; Hurka, H.; Shmakov, A.; Oyuntsetseg, B.; Zerdoner Calasan, A.; et al. The phylogeographic history of Krascheninnikovia reflecs the development of the dry steppes and semy-deserts in Eurasia. Sci. Rep. 2021, 11, 6645. [Google Scholar] [CrossRef]
  61. Volkova, P.; Herden, T.; Friesen, N. Genetic variation in Goniolimon speciosum (Plumbaginaceae) reveals a complex history of steppe vegetation. Bot. J. Linn. Soc. 2017, 184, 113–121. [Google Scholar]
  62. Hurka, H.; Friesen, N.; Bernhardt, K.-G.; Neuffer, B.; Smirnov, S.V.; Shmakov, A.I.; Blattner, F.R. The Eurasian steppe belt: Status quo, origin and evolutionary history. Turczaninowia 2019, 22, 5–71. [Google Scholar]
  63. Török, P.; Neuffer, B.; Heilmeier, H.; Bernhardt, K.-G.; Wesche, K. Climate, landscape history and management drive Eurasian steppe biodiversity. Flora 2020, 271, 151685. [Google Scholar] [CrossRef]
  64. Seregin, A.P. (Ed.) Moscow Digital Herbarium: Electronic Resource. Moscow State University: Moscow, Russia. Available online: https://plant.depo.msu.ru/ (accessed on 1 April 2022).
  65. Kovtonyuk, N.K.; Han, I.; Gatilova, E.; Friesen, N. Genus Allium in CSBS Digital Herbarium. BIO Web Conf. 2020, 24, 00042. [Google Scholar] [CrossRef]
  66. Thiers, B. Index Herbariorum: A Global Directory of Public Herbaria and Associated Staff. New York Botanical Garden’s Virtual Herbarium. Available online: http://sweetgum.nybg.org/science/ih/ (accessed on 15 January 2022).
  67. Blattner, F.R. Direct amplification of the entire ITS region from poorly preserved plant material using recombinant PCR. Biotechnology 1999, 27, 1180–1185. [Google Scholar] [CrossRef]
  68. White, T.J.; Bruns, T.; Lee, S.; Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: A Guide to Methods and Applications; Academic Press: New York, NY, USA, 1990; pp. 315–322. [Google Scholar] [CrossRef]
  69. Shaw, J.E.B.; Lickey, E.E.; Schilling, E.E.; Small, R.L. Comparison of whole chloroplast genome sequences to choose noncoding regions for phylogenetic studies in angiosperms: The tortoise and the hare III. Am. J. Bot. 2007, 94, 275–288. [Google Scholar] [CrossRef] [Green Version]
  70. Thompson, J.D.; Gibson, T.J.; Plewniak, F.; Jeanmougin, F.; Higgins, D.G. The Clustal X window interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997, 25, 4876–4882. [Google Scholar] [CrossRef] [Green Version]
  71. Kumar, S.; Stecher, G.; Tamura, K. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 2016, 33, 1870–1874. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  72. Swofford, D.-L. PAUP*: Phylogenetic Analysis Using Parsimony (* and Other Methods), Version 4; Sinauer Associates: Sunderland, MA, USA, 2002.
  73. Kluge, A.G.; Farris, J.S. Quantitative phyletics and the evolution of anurans. Syst. Zool. 1969, 18, 1–32. [Google Scholar] [CrossRef]
  74. Felsenstein, J. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 1985, 39, 783–791. [Google Scholar] [CrossRef] [PubMed]
  75. Ronquist, R.; Huelsenbeck, J.P. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 2003, 19, 1572–1574. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  76. Darriba, D.; Taboada, G.L.; Doallo, R.; Posada, D. jModelTest 2: More models, new heuristics and parallel computing. Nat. Methods 2012, 9, 77. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  77. Smirnov, Y.A. Accelerated method of study of somatic chromosomes of fruit plants. Tsitologiya 1968, 10, 1601–1602. (In Russian) [Google Scholar]
  78. Altinordu, F.; Peruzzi, L.; Yu, Y.; He, X. A tool for the analysis of chromosomes: KaryoType. Taxon 2016, 65, 586–592. [Google Scholar] [CrossRef]
  79. Pfosser, A.; Amon, A.; Lelley, T.; Heberle-Bors, E. Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines. Cytometry 1985, 21, 387–393. [Google Scholar] [CrossRef]
  80. Doležel, J.; Greilhuber, J.; Lucretti, S.; Meister, A.; Lysák, M.A.; Nardi, L.; Obermayer, R. Plant genome size estimation by flow cytometry: Inter-laboratory comparison. Ann. Bot. 1998, 82, 17–26. [Google Scholar] [CrossRef] [Green Version]
Plants 11 01465 g001 550
Figure 1. The geographic location of collected accessions and distribution of A. pallasii and A. caricifolium.
Figure 1. The geographic location of collected accessions and distribution of A. pallasii and A. caricifolium.
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Plants 11 01465 g002 550
Figure 2. Boxplot analysis of different morphological character (in mm). (A): Carpel length, (B): Pedicel length, (C): Length of filaments outside tepals, (D): Filaments length in total, (E): Tepal length, (F): Scape length. The sizes range between 0–30 mm (A–E), the scape length is between 0 and 400 mm (F). A. pallasii is shown in green, A. caricifolium in blue.
Figure 2. Boxplot analysis of different morphological character (in mm). (A): Carpel length, (B): Pedicel length, (C): Length of filaments outside tepals, (D): Filaments length in total, (E): Tepal length, (F): Scape length. The sizes range between 0–30 mm (A–E), the scape length is between 0 and 400 mm (F). A. pallasii is shown in green, A. caricifolium in blue.
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Plants 11 01465 g003 550
Figure 3. PCA analysis of morphological characters, shown in Table 1.
Figure 3. PCA analysis of morphological characters, shown in Table 1.
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Plants 11 01465 g004 550
Figure 4. Inflorescences of the Allium pallasii: (A)—Am588 (Photo S. Smirnov); (B)—Am780; (C)–Am189; (D). Am607; Allium caricifolium: (E)—Am708; (F)—Am580; (G)—Am494. (All photos, except Am588, by N. Friesen, origin of the accessions in Appendix A). Accessions Am607 (D) and Am708 (E) were used to determine relative DNA amount.
Figure 4. Inflorescences of the Allium pallasii: (A)—Am588 (Photo S. Smirnov); (B)—Am780; (C)–Am189; (D). Am607; Allium caricifolium: (E)—Am708; (F)—Am580; (G)—Am494. (All photos, except Am588, by N. Friesen, origin of the accessions in Appendix A). Accessions Am607 (D) and Am708 (E) were used to determine relative DNA amount.
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Plants 11 01465 g005 550
Figure 5. Generalized nrITS tree of the third evolutionary line of genus Allium. Numbers by nodes represent bootstrap support (1000 replicates) and Bayesian probabilities. Roman numerals (I, II, and III) designate clades of three evolutionary lines. The joint presence of Bayesian probabilities over 0.98 and bootstrap support over 95% is indicated with a black dot. Allium pallasii and A. caricifolium clades are sister. Marked red in the tree. For the origin of samples without GenBank accession numbers, see Appendix A.
Figure 5. Generalized nrITS tree of the third evolutionary line of genus Allium. Numbers by nodes represent bootstrap support (1000 replicates) and Bayesian probabilities. Roman numerals (I, II, and III) designate clades of three evolutionary lines. The joint presence of Bayesian probabilities over 0.98 and bootstrap support over 95% is indicated with a black dot. Allium pallasii and A. caricifolium clades are sister. Marked red in the tree. For the origin of samples without GenBank accession numbers, see Appendix A.
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Plants 11 01465 g006 550
Figure 6. Combined nrITS and plastid (trnL-rpl32 and trnQ-rps16) tree of A. sect. Pallasia. Species from A. sect. Codonoprasum has been used as an outgroup. Numbers by nodes represent bootstrap support (1000 replicates) and Bayesian probabilities. The joint presence of Bayesian probabilities over 0.98 and bootstrap support over 95% is indicated with a black dot. The origin of samples is seen in Appendix A.
Figure 6. Combined nrITS and plastid (trnL-rpl32 and trnQ-rps16) tree of A. sect. Pallasia. Species from A. sect. Codonoprasum has been used as an outgroup. Numbers by nodes represent bootstrap support (1000 replicates) and Bayesian probabilities. The joint presence of Bayesian probabilities over 0.98 and bootstrap support over 95% is indicated with a black dot. The origin of samples is seen in Appendix A.
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Plants 11 01465 g007 550
Figure 7. (A)—Idiogram of Allium pallasii, based on 36 metaphase plates; (B)—metaphase plate of Am804, (C)—Idiogram of A. caricifolium based on five metaphase plates; (D)–metaphase plate of Am1246. Arrows point to satellites. Roman numbers show the numbering of chromosome pairs.
Figure 7. (A)—Idiogram of Allium pallasii, based on 36 metaphase plates; (B)—metaphase plate of Am804, (C)—Idiogram of A. caricifolium based on five metaphase plates; (D)–metaphase plate of Am1246. Arrows point to satellites. Roman numbers show the numbering of chromosome pairs.
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Table
Table 1. Comparison of morphological characters of A. pallasii (20 individuals) and A. caricifolium (22 individuals). Quantitative characters are expressed in mm and presented as mean ± standard deviation (extreme values in brackets).
Table 1. Comparison of morphological characters of A. pallasii (20 individuals) and A. caricifolium (22 individuals). Quantitative characters are expressed in mm and presented as mean ± standard deviation (extreme values in brackets).
CharacterA. pallasiiA. caricifolium
Scape length194.7 ± 55.3 (125–290)275 ± 63 (200–388)
Pedicel length12.9 ± 3.7 (7.5–19)10.4 ± 1.6 (6.5–14)
Tepals length2.9 ± 0.1 (2.1–3.3)3 ± 0.2 (2.0–4.0)
The ratio of tepals length/pedicels length1.11.2
Length of filaments outside the tepals 0.1 ± 0.1 (0)1 ± 0.1 (0.7–1.1)
Filaments length in total2.9 ± 0.1 (2.6–3)4 ± 0.1 (3.8–4.3)
The ratio filaments length outside the tepals/total length1.021.25
Carpel length2.8 ± 0.1 (2–3.3)3.9 ± 0.1 (2.9–4.6)
Flowers per inflorescence36.8 ± 17.5 (22–75)52.8 ± 12.9 (37–66)
Table
Table 2. Karyo-morphometric parameters of A. pallasii accessions in µm.
Table 2. Karyo-morphometric parameters of A. pallasii accessions in µm.
Pair No. TAL (µm)RL %LA (µm)SA (µm)SatCI %Type
16.6 ± 0.815.1 ± 0.23.5 ± 0.43.0 ± 0.5046.3 ± 1.4m
26.2 ± 0.714.3 ± 0.23.4 ± 0.32.9 ± 0.4046.1 ± 2.0m
35.9 ± 0.713.6 ± 0.23.4 ± 0.42.6 ± 0.4043.4 ± 1.1m
45.6 ± 0.912.9 ± 0.43.2 ± 0.52.4 ± 0.4043.2 ± 1.1m
55.4 ± 0.712.3 ± 0.13.1 ± 0.32.3 ± 0.5042.8 ± 2.6m
65.0 ± 0.611.5 ± 0.32.8 ± 0.22.2 ± 0.4044.5 ± 2.4m
74.7 ± 0.610.8 ± 0.12.7 ± 0.42.1 ± 0.3043.8 ± 1.5m
84.2 ± 0.69.6 ± 0.12.3 ± 0.21.9 ± 0.4044.4 ± 2.6m
Abbreviations: TAL total absolute length; RL relative length; LA long arm; SA short arm; Sat satellite; CI centromeric index; Type chromosome nomenclature according to [32,33]; TKL total karyotype diploid length. MCA mean centromeric asymmetry; KCI Karyotype centromeric index. A. pallasii: TKL = 87.1 ± 11.6; MCA = 11.4 ± 2.5; CItotal = 44.3 ± 1.3.
Table
Table 3. Karyo-morphometric parameters of A. caricifolium in µm.
Table 3. Karyo-morphometric parameters of A. caricifolium in µm.
Pair No. TAL (µM)RL %LA (µM)SA (µM)SatCI %Type
17.2 ± 1.413.9 ± 2.54.3 ± 0.72.9 ± 0.9040.7 ± 8.7m
26.7 ± 1.213.1 ± 2.13.7 ± 0.63.0 ± 1.1044.6 ± 11.4m
36.6 ± 1.112.9 ± 1.93.9 ± 0.72.8 ± 0.5041.6 ± 4.0m
46.5 ± 0.312.5 ± 1.13.9 ± 0.32.5 ± 0.3039.1 ± 4.5m
56.4 ± 0.512.4 ± 0.93.7 ± 0.32.7 ± 0.5041.9 ± 5.9m
66.4 ± 0.812.4 ± 1.23.7 ± 0.41.8 ± 0.8127.4 ± 8.1m
75.9 ± 1.311.4 ± 2.03.4 ± 0.72.5 ± 0.6042.4 ± 5.6m
85.9 ± 1.011.4 ± 2.53.7 ± 0.72.2 ± 0.6236.7 ± 7.6m
Abbreviations: TAL total absolute length; RL relative length; LA long arm; SA short arm; Sat satellite; CI centromeric index; Type chromosome nomenclature according to [32,33]; TKL total karyotype diploid length. MCA mean centromeric asymmetry; KCI Karyotype centromeric index. A. caricifolium: TKL = 103.2 ± 5; MCA = 18.9 ± 4.8; CItotal = 40.5 ± 5.3.
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Friesen, N.; Grützmacher, L.; Skaptsov, M.; Vesselova, P.; Dorofeyev, V.; Luferov, A.N.; Turdumatova, N.; Lazkov, G.; Smirnov, S.V.; Shmakov, A.I.; et al. Allium pallasii and A. caricifolium—Surprisingly Diverse Old Steppe Species, Showing a Clear Geographical Barrier in the Area of Lake Zaysan. Plants 2022, 11, 1465. https://doi.org/10.3390/plants11111465

AMA Style

Friesen N, Grützmacher L, Skaptsov M, Vesselova P, Dorofeyev V, Luferov AN, Turdumatova N, Lazkov G, Smirnov SV, Shmakov AI, et al. Allium pallasii and A. caricifolium—Surprisingly Diverse Old Steppe Species, Showing a Clear Geographical Barrier in the Area of Lake Zaysan. Plants. 2022; 11(11):1465. https://doi.org/10.3390/plants11111465

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Friesen, Nikolai, Lisa Grützmacher, Mikhail Skaptsov, Polina Vesselova, Vladimir Dorofeyev, Alexander N. Luferov, Nazgul Turdumatova, Georgii Lazkov, Sergei V. Smirnov, Alexander I. Shmakov, and et al. 2022. "Allium pallasii and A. caricifolium—Surprisingly Diverse Old Steppe Species, Showing a Clear Geographical Barrier in the Area of Lake Zaysan" Plants 11, no. 11: 1465. https://doi.org/10.3390/plants11111465

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