Analysis of genetic variation in rare endemic species Oxytropis chankaensis Jurtz. (Fabaceae) using RAPD markers

The method of polymerase chain reaction with random primers (RAPD) was used to assess genetic variation and population differentiation in the rare endemic plant Oxytropis chankaensis Jurtz. (Fabaceae). DNA samples from plants of two isolated populations were compared at 133 loci detected by use of ten primers. Both populations examined were characterized by high polymorphism levels (P95 = 72.9%, A = 1.92 and P95 = 74.4%, A = 1.88, respectively). They were also statistically significantly different in the frequencies of most of the amplicons. For each of the plants, unique multilocus RAPD phenotype was established using 17 to 20 RAPD markers. Diagnostic markers were not revealed. The populations were poorly differentiated. On average, the between-population component accounted for about 8% of the variation, while 92% of the variation was detected within populations. High variation along with the low degree of differentiation characteristic of two most geographically remote populations of O. chankaensis can have several explanations, among which a polyploid origin of the species seems to be most important.     

Analysis of Genetic Variation in Rare Endemic Species Oxytropis chankaensis Jurtz. (Fabaceae) Using RAPD Markers

The method of polymerase chain reaction with random primers (RAPD) was used to assess genetic variation and population differentiation in the rare endemic plant Oxytropis chankaensis Jurtz. (Fabaceae). DNA samples from plants of two isolated populations were compared at 133 loci detected by use of ten primers. Both populations examined were characterized by high polymorphism levels (P ₉₅ = 72.9%; A = 1.92 and P ₉₅ = 74.4%; A = 1.88, respectively). They were also statistically significantly different in the frequencies of most of the amplicons. For each of the plants, unique multilocus RAPD phenotype was established using 17 to 20 RAPD markers. Diagnostic markers were not revealed. The populations were poorly differentiated. On average, the between-population component accounted for about 8% of the variation, while 92% of the variation was detected within populations. High variation along with the low degree of differentiation characteristic of two most geographically remote populations of O. chankaensis can have several explanations, among which a polyploid origin of the species seems to be most important.     

Amplified Fragment Length Polymorphism (AFLP) reveals no genetic divergence of the Eastern Alpine endemic Oxytropis campestris subsp. tiroliensis (Fabaceae) from widespread subsp. campestris

Applying Amplified Fragment Length Polymorphism, we explored genetic differences between widespread Oxytropis campestris subsp. campestris and O. campestris subsp. tiroliensis, a presumed glacial relict restricted to a small area along the main chain of the Eastern Alps. We could not find genetic differences between the two taxa. Neither do the morphological characters given in the literature discriminate between them. Therefore Oxytropis campestris subsp. tiroliensis is unlikely a glacial relict that survived Pleistocene glaciations on nunataks, but rather a genetically insignificantly differentiated phenotype that arose in the course of postglacial recolonisation. There is no phylogeographical structure in O. campestris s.l. in the Alps most probably due to the fact that the taxon did not survive the cold stages of the Pleistocene in the interior of the Alps but immigrated to that region at a later date.     

Seed morphology and its systematic implications for genus Oxytropis DC. (Fabaceae)

In the present study, seeds of 13 Oxytropis DC. species (including two latest synonyms) classified in subg. Oxytropis (sect. Dolichocarpon, sect. Chrysantha and sect. Eumorpha), subg. Euoxytropis (sect. Orobia) and subg. Phacoxytropis (sect. Protoxytropis, sect. Janthina and sect. Mesogaea) from Turkey were examined using light and scanning electron microscopy to evaluate the taxonomic relevance of macro- and micro-morphological seed characters. As a result of the study, species-specific characters have been determined. Seeds exhibit variation in size, shape, surface sculpturing pattern, hilum position and weight. Seeds ranged from 1.61 to 4.04 mm in length and from 1.02 to 2.56 mm in width. Five different seed shapes were recognised, as prolonged semielliptic, reniform, prolonged reniform, quadratic and cardiform, with a length to width ratio ranging from 1.02 ± 0.08 to 1.75 ± 0.27. Rugulate, rugulate–reticulate and lophate sculpturing patterns were observed in the studied species. According to the results, seed characters, such as the size (length, width and the length/width ratio), the shape, the surface sculpturing and the weight appear to have low taxonomic value in distinguishing subgenera, sections and species.     

Molecular taxonomy: description of a cryptic Petitia species (Polychaeta: Syllidae) from the island of Mahé (Seychelles, Indian Ocean) using RAPD markers and ITS2 sequences

Petitia occulta sp. n. provides an example demonstrating that a morphologically indistinguishable species taxon can be described by molecular markers alone. Phenotypically the polychaete belongs to Petitia amphophthalma Siewing, 1956, one of the numerous meiofaunal species that have previously been regarded as cosmopolitan. Genotypically, however, by means of randomly amplified polymorphic DNA (RAPD)–polymerase chain reaction (PCR) (a DNA-fingerprinting technique) the population on the Indo-Pacific island of Mahé can be identified as a distinct taxonomic entity as: (1) genetic distance values between the Petitia from the Seychelles and from other sites (Mediterranean, Canary Islands) are relatively great; (2) differently generated cluster analyses of genetic distances produce phenograms revealing a distinctly separate clade (bootstrap 100%) for the Seychelles animals; and (3) the Seychelles animals are characterized by eight monomorphic diagnostic (i.e. autapomorphic) DNA-fragments. The RAPD procedure in this case requires the preparation of the whole organisms and, thus, no holotype is available. Instead the RAPD band-pattern with the diagnostic markers of one individual obtained with six primers, isolated DNA of one individual, and syntypes in the form of fixed, complete individuals are deposited. Species differentiation is confirmed by genomic DNA sequencing (ITS2).     

Phylogeny and biogeography of Alyssum (Brassicaceae) based on nuclear ribosomal ITS DNA sequences

The genus Alyssum consists of about 195 species native to Europe, Asia and northern Africa. All species were assigned to six sections. Previous molecular phylogeny studies indicate that Alyssum is polyphyletic. However, the divergence time and dispersal of the genus are not well studied. In this study, the phylogenetic relationships within the genus Alyssum were studied with nrDNA ITS sequences obtained from five sections. The divergence time was estimated by fossil calibration and the biogeography was examined by spread analysis. The phylogeny indicated two main lineages: lineage 1 includes the section of Alyssum, Gamosepalum and Psilonema; lineage 2 includes the section of Odontarrhena, Meniocus and Clypeola. The phylogenetic relationship was not congruent with the previous sectional classifications. The age of Alyssum was dated to the upper Miocene. Molecular data suggested the diversification of Alyssum in Mediterranean areas and wide-ranging distribution such as North Africa, eastward into Central Asia and immigration into North America. Climatic aridification and arid/semiarid areas established in the Pliocene/Pleistocene could have provided favourable conditions for the migration and diversification of Alyssum.     

Genetic variation in six species of the genus Oxytropis DC. (Fabaceae) from Kamchatka Peninsula

Using the isozyme analysis, genetic variations in six species of the genus Oxytropis DC. (Fabaceae) from Kamchatka was assessed. It was demonstrated that diploid species from the section Arctobia were characterized by a low level of variations typical of endemic plant species. At the same time, polyploid species from the Orobia section demonstrated very high values of the heterozygosity parameters (H _o varied from 0.200 to 0.274). It has been suggested that the level of polymorphism of the oxytropes from Kamchatka was shaped as a result of the interaction of a number of factors, among which the most important are the ecological confinedness of the species, the specific features of the reproductive system, and gene drift. In the species of Orobia section, it is also the presence of the polyploid genome.     

Phylogeny and taxonomy of the Pyrenaria complex (Theaceae) based on nuclear ribosomal ITS sequences

The generic limits in the Pyrenaria complex are discussed based on morphological and molecular evidence. The phylogeny of the Pyrenaria complex was reconstructed using parsimony and Bayesian analyses of sequence data from the nuclear internal transcribed spacer (ITS) region. The Pyrenaria complex is shown to be monophyletic and is sister to the Camellia–Polyspora clade. None of the previously recognized genera (including Pyrenaria, Tutcheria, Parapyrenaria and Sinopyrenaria) is monophyletic, and the species of these genera are scattered and nested within the Pyrenaria complex, suggesting that the four genera should be combined as Pyrenaria sensu lato. This study indicates a close relationship among the genera within the Pyrenaria complex, Camellia and Polyspora, and reveals that Camellia has a closer relationship with Polyspora than with the Pyrenaria complex.     

Inheritance of oilseed rape (Brassica napus) RAPD markers in a backcross progeny with Brassica campestris

Different cultivars/transgenic lines of oilseed rape (Brassica napus) were crossed (as females) with different cultivars/populations of Brassica campestris. All cross combinations produced seed, with an average seed set per pollination of 9.8. Backcrossing of selected interspecific hybrids (as females) to B. campestris resulted in a much lower seed set, average 0.7 seed per pollination. In the single backcross progeny where a large enough population (92 plants) was obtained for analysis, 33 B. napus specific RAPD markers were investigated to determine the extent of transfer of oilseed rape genetic material into this population. Markers were transferred to the backcross generation with frequencies ranging from 26% to 91%. Almost all of the markers (30/33) were transferred in a frequency not significantly different from 50%. Analysis of the pairwise segregation of markers revealed that 23 markers could be assigned to six linkage groups, most probably reflecting six B. napus C-chromosomes. The presence of backcross plants with recombinant genotypes suggests that complex genetic processes can take place during interspecific hybridisation and backcrossing in these Brassica species. The implications of our results for the possible choice of integration sites of transgenes in oilseed rape are discussed.     

Comparison of RAPD and ISSR markers for assessment of genetic diversity among endangered rare Dalbergia oliveri (Fabaceae) genotypes in Vietnam

Dalbergia oliveri is a leguminous tree of the Fabaceae family. This species is popular and valuable in Vietnam and is currently listed on the Vietnam Red List and on the IUCN Red List as endangered. Two PCR techniques using RAPD and inter-simple sequence repeat (ISSR) markers were used to make a comparative analysis of genetic diversity in this species. Fifty-six polymorphic primers (29 RAPD and 27 ISSR) were used. The RAPD primers produced 63 bands across 35 genotypes, of which 24 were polymorphic. The number of amplified bands varied from one to four, with a size range from 250 to 1400 bp. The percentage polymorphism ranged from 0 to 75. Amplification of genomic DNA of the 35 genotypes, using ISSR analysis, yielded 104 fragments, of which 63 were polymorphic. The number of amplified fragments using ISSR primers ranged from one to nine and varied in size from 250 to 1500 bp. The percentage polymorphism ranged from 0 to 100. ISSR markers were relatively more efficient than RAPDs. The mental test between two Jaccard's similarity matrices gave r ≥0.802, showing good fit correlation between ISSRs and RAPDs. Clustering of isolates remained more or less the same for RAPDs compared to combined RAPD and ISSR data. The similarity coefficient ranged from 0.785 to 1.000, 0.698 to 0.956 and 0.752 to 0.964 with RAPD, ISSR, and the combined RAPD-ISSR dendrogram, respectively.     

Phylogenetic relationships of Oxytropis chankaensis Jurtz. and Oxytropis oxyphylla (Pall.) DC. (Fabaceae) inferred from the data of sequencing of the ITS region of the nuclear ribosomal DNA operon and intergenic spacers of the chloroplast genome

The autotetraploid Oxytropis chankaensis Jurtz. is the only representative of the section Baicalia (subgenus Oxytropis, Fabaceae) in Primorskii Krai, and its range is restricted to the sandy belt along the western coast of the Khanka Lake. The type species of the section Baicalia is O. oxyphylla (Pall.) DC., and O. chankaensis is regarded by some authors as its synonym. Analysis of the ITS sequences showed that O. chankaensis is phylogenetically closely related to the species O. oxyphylla and O. racemosa Turcz. from the section Baicalia and also to the species O. anertii Nakai., O. ciliata Turcz., and O. inschanica H. C. Fu & S. H. Cheng from the section Xerobia, but this analysis failed to resolve the phylogenetic relationships of the species within the genus Oxytropis. The analysis of the trnH-psbA, trnL-trnF, trnS-trnG, and petG-trnP regions of cpDNA revealed molecular differences between O. chankaensis and O. oxyphylla that are indicative of considerable divergence of their chloroplast genomes. The ancestral lineages of these species diverged nearly 2 million years ago, which confirms the taxonomic validity of O. chankaensis. Taking into account the ecological specificity of O. chankaensis and the present-day distribution range of O. oxyphylla, it can be assumed that they descended from one ancestral maternal lineage that gave rise to the species of the section Baicalia.     

Genetic diversity of Oxytropis section Xerobia (Fabaceae) in one of the centres of speciation

Genetica - The genetic diversity and phylogenetic relationships of Oxytropis caespitosa, O. grandiflora, O. eriocarpa, O. mixotriche, O. nitens,...     

青藏高原六种棘豆属植物的染色体数目及核型报道

对6种棘豆属植物（Oxytropis）的13个居群样进行细胞学研究,其中黄花棘豆（O．ochrocephala）、绢毛棘豆（O．tatarica）、甘肃棘豆（O．kansuensis）和铺地棘豆（0．humifusa）为首次报道染色体数目（2n=16）和核型;在胀果棘豆（O．stracheyana）中（2n=48）首次发现B染色体。现有的细胞学资料表明：棘豆属植物中多倍体占总报道数的58％,这说明多倍化在本属植物的进化过程中起着非常重要的作用,但青藏高原仅有一种植物发现多倍体,多倍化并不占主导地位,而主要表现为二倍体水平上的结构变异,即核型不对称性的变化。     

Allozyme variation in relation to morphology and taxonomy inMedicago sect.Spirocarpos subsect.Intertextae (Fabaceae)

Medicago intertexta andM. ciliaris have been controversially recognized as separate species. The only reliable diagnostic character, gland-tipped trichomes on the fruits inM. ciliaris, is controlled by presence of a single dominant allele, and such one-character taxonomies are debatable. Contributing to the difficulty,M. muricoleptis andM. granadensis, the other two species ofMedicago sectionSpirocarpos subsectionIntertextae, are sometimes confusingly similar toM. intertexta or to each other. Allozyme differences provided 95% verification of the suitability of the gland-tipped trichome character for separatingM. intertexta andM. ciliaris, thus corroborating their recognition as separate taxa. Several measures of allozyme variation indicated thatM. intertexta is more polymorphic than its sister species. Heterozygosity was also highest inM. intertexta, suggestive of a higher outcrossing rate, which is also consistent with larger floral size. Heterozygosity ofM. intertexta was concentrated in Sicily and nearby countries. Taxonomic difficulties in identifying SicilianM. intertexta are well known, and may be the result of interspecific hybridization and introgression.Medicago muricoleptis differed from the above two species in the frequency of several alleles, whileM. granadensis possessed numerous unique alleles consistent with its complete absence of genetic exchange with the other three substantially interfertile species.     

Development and characterisation of nine polymorphic microsatellite markers for Tephrosia calophylla Bedd. (Fabaceae)

Tephrosia calophylla Bedd. (Fabaceae) is an endangered tropical plant endemic to southwestern Ghats, India. The objective of this study was to contribute to the characterisation of the diversity of this rare species, which is necessary for its future conservation. Accordingly, microsatellite markers were designed, and their ability to detect polymorphisms was determined. Nine microsatellite markers were developed using genomic libraries, and all of the markers were successfully amplified in 42 individuals. Three to nine alleles per locus were observed, and the heterozygosity of the loci ranged from 0.381 to 0.905. The nine newly developed polymorphic markers recognise a sufficient number of varying loci to perform further studies on the conservation and breeding of this medicinal cultivar.     

甘肃省冬虫夏草菌的RAPD和ITS序列分析

目的对甘肃省冬虫夏草分离菌株进行鉴定和系统发育研究。方法采用随机扩增多态性DNA（RAPD）分析和核糖体（rDNA）内部转录间隔区（ITS）序列分析。结果共分离到18个菌株,菌株间的遗传相似性系数为0.623～1.000,遗传分化距离为0.000～0.018,每个菌株的序列与中国被毛孢的序列一致性均高于98%且18个菌株（G＋C）%的差别最大值是4.7%。结论 18个分离菌株均为中国被毛孢;多样性分析表明,不同区域样本间的遗传分化较大;同一地点样本间的遗传分化很小;同一材料不同部位获得的菌株在RAPD标记上差异无统计学意义,两技术对相同的供试菌株所反映的遗传分化不尽相同,表明两技术用于冬虫夏草遗传多样性和种类鉴定上各有它们的优势和具体选择。