Allozyme diversity in the tetraploid endemic Thymus loscosii (Lamiaceae)

BACKGROUND AND AIMS: Thymus loscosii (Lamiaceae) is a tetraploid perennial species endemic to the Ebro river basin (north-eastern Spain), which is included in the National Catalogue of Endangered Species. It is a tetraploid species (2n = 54), presumably an autotetraploid originated by the duplication of a 2n = 28 genome and the subsequent loss of two chromosomes. Allozyme electrophoresis was conducted to survey the levels and distribution of genetic diversity and to test the previous autopolyploid hypothesis for its origin. In addition, both in situ and ex situ conservation measures are proposed. METHODS: Eight populations were sampled for analysis by standard methods of starch gel electrophoresis, and six putative enzymatic loci were resolved (five consistently and one only partially). KEY RESULTS: Banding patterns exhibited no evidence of fixed heterozygosity and showed both balanced and unbalanced heterozygotes. In addition, most individuals showed a pattern consistent with the presence of three or four alleles at a single locus. High levels of genetic variability were found at population level (P = 85 %, A = 3.0, He = 0.422), in addition to a trend of an excess of heterozygotes. CONCLUSIONS: Allozyme data support the hypothesis that T. loscosii is an autotetraploid, and the high number of alleles at some loci may be due to repeated polyploidization events. The high values of genetic variation found in this species agree with those expected for tetraploids. The excess of heterozygotes may be due to some barriers to inbreeding (e.g. occurrence of gynodioecy) and/or selection for heterozygosity.     

Genetic diversity and population structure of the serpentine endemic Calystegia collina (Convolvulaceae) in northern California

We used enzyme electrophoresis to evaluate genetic diversity in 32 populations of Calystegia collina, a clonal plant species endemic to serpentine outcrops in northern California (USA). Of 34 loci examined 56% were polymorphic, but on average only 17% were polymorphic within local populations. Neither the total number of alleles nor the number of multilocus genotypes differed significantly between populations in small vs. large serpentine outcrops. Genetic and geographic distances between populations were positively correlated, but this relationship was not significantly affected by the isolation of serpentine outcrops. Populations were highly differentiated (F(st) = 0.417) and little genetic variation was explained by geographic region or serpentine outcrop.Observed heterozygosity within populations almost always exceeded Hardy-Weinberg expectations. In many populations, all 30 sample ramets were uniformly heterozygous at one or more loci yet were genetically variable at other loci. These results imply that many C. collina populations originate from one or a few genetic founders, with little recruitment from seeds. Genetic variation within uniformly heterozygous populations must be the product of multiple, closely related founders or somatic mutations within the population. We conclude that vegetative reproduction, perhaps coupled with somatic mutation, helps maintain genetic diversity in these isolated but long-lived populations.     

Genetic diversity of the narrow endemic Astragalus oniciformis (Fabaceae)

Astragalus oniciformis Barneby is a narrow endemic xerophyte of the upper Snake River Plain of central Idaho, USA, where it inhabits stabilized, aeolian sand deposits and previously burned, sandy sites over Quaternary basalt flows. The objective of this study was to determine the levels and distribution of genetic differentiation within and among populations of A. oniciformis. Fifteen individuals from each of eight populations, chosen from throughout the range of the species, were selected based on accessibility, density of individuals, and large population size. Inter-simple sequence repeats were chosen as the marker to assess genetic differentiation. The two primers selected yielded 40 polymorphic loci in A. oniciformis. In an analysis of molecular variance, 88.69% of the variation was significantly attributed to variation within populations. High gene flow (N(m) = 3.91-3.93; SD = 0.01) and a low percentage deviation from Hardy-Weinberg equilibrium due to population subdivision (G(ST) = 0.113-0.1134; SD = 0.0002) were found among sampled populations. These results suggest that current threats to this species, including changing fire patterns and habitat loss from grazing disturbance, have not yet affected the genetic diversity of this species. Preservation of large populations and smaller, intervening, dispersed patches will help preserve the genetic integrity and the genetic diversity found in A. oniciformis.     

Isozyme diversity in Iris Cristata and the threatened glacial endemic I. Lacustris (Iridaceae)

Iris cristata and I. lacustris differ markedly in geographic distribution, glacial history of current ranges, and ecology. We hypothesized that I. cristata, a widespread species of unglaciated regions of eastern North America, would exhibit genetic diversity typical of other widespread plant species, whereas the threatened I. lacustris, which occupies glaciated habitats on Great Lakes shorelines, would display little genetic variation. Iris lacustris lacked detectable polymorphisms in 18 isozyme loci, although we found evidence of possible incomplete gene silencing in four additional loci in some populations. In contrast, I. cristata was polymorphic at 73% of 15 loci examined, with an average of three alleles per locus. Genetic diversity (H_e) was 0.231. All species-level and population-level estimates of genetic diversity were higher than averages for plants having comparable life history traits. Nearly 98% of the total genetic diversity in I. cristata was apportioned within populations, and heterozygosity and fixation estimates suggest a high level of outcrossing in this species (t = 1.265). The long-lived perennial habit and high outcrossing rate in stable populations are proposed as factors contributing to high genetic diversity in I. cristata. The data are consistent with an hypothesis of a recent origin of I. lacustris from a very limited I. cristata gene pool exacerbated by repeated bottlenecks and founder effects as I. lacustris populations were displaced by lake-level changes over the past 11 000 yr.     

Genetic diversity in the top onion,Allium ×proliferum (Alliaceae), analysed by isozymes

Polymorphisms for six enzyme systems (GPI, IDH, PDG, PGM, SKD, and TPI) were analysed in the top onion,Allium ×proliferum. Five multilocus isozyme genotypes were found. The banding patterns of top onions were compared with those ofA. ×wakegi, A. cepa, A. fistulosum, A. altaicum, and artificial hybrids between these three species. One top onion type and one artificial hybrid had identical banding patterns. Shallots andA. altaicum, the wild progenitor ofA. fistulosum, cannot be distinguished from the common onion andA. fistulosum, respectively; these species are also potential contributors to the top onion's gene pool.     

Genetic variation relevant for the conservation of the narrow endemic Senecio carbonensis (Asteraceae) from the southern Andes

We studied morphological variation, reproductive biology and genetic structure of Senecio carbonensis, a narrow endemic from high elevations of the southern Andes, and compared it to Senecio peteroanus, a closely related widespread congeneric species. Using ANOVAs and discriminant analysis we showed that populations of S. carbonensis were more similar to each other in their morphology, had comparatively limited reproductive capacity and presented lower plant density and more reduced plant cover than populations of S. peteroanus. Similar high genetic variation was found at the population and species level in both species, based on isozyme variation at 14 and 11 putative gene loci that were resolved in S. carbonensis and S. peteroanus, respectively. The two species were genetically distinct. However, the small genetic distance between populations of each species suggests that either they were recently founded and that genetic drift did not have time to promote divergence, or that its effects are masked by significant current gene flow. These results show that ecological and reproductive characteristics of S. carbonensis may limit its extension and abundance. In contrast, high genetic variation at the population level is probably favored by self‐incompatibility. Despite the restricted geographical distribution and low plant density of S. carbonensis populations, genetic characteristics do not seem to be limiting their long‐term persistence. This information suggests that populations of S. carbonensis are not at risk of extinction as a result of genetic factors, although it is important to study and monitor population dynamics to further assess the degree of recruitment through time.     

Genetic diversity in the threatened insular endemic plantAster asa-grayi (Asteraceae)

Genetic diversity in an insular endemic plantAster asa-grayi was examined using enzyme electrophoresis. Distribution ofA. asa-grayi is restricted to only four subtropical islands of Japan, and this species is listed as vulnerable to extinction in the Red Data Book of Japanese wild plants. A total of 161 individuals were sampled from five populations on four islands. Genetic diversity values at the population level were very low, compared to other plant species with a similar life history. Genetic variability at the species level is comparable to the mean value of endemic species. Genetic differentiation among populations is extremely high (G_ST= 0.71), indicating that the gene flow among populations is highly impeded, and pollen and seed dispersal is limited due to the pollinators and the seed morphology. This is because the four islands are geographically isolated. Fixation indices suggested that most populations do not randomly cross. To conserve the genetic diversity of the species, artificial crossings among different island populations are necessary.     

A new species of Allium (Alliaceae) from Dalmatia,Croatia

Allium croaticum , a new species from the island of Vis in Central Dalmatia (Croatia) is described and illustrated. Its relationships with allied species belonging to the A. stamineum group (Allium section Codonoprasum) are discussed. It is a diploid species (2n = 16), colonizing calcareous screes and flowering in early summer. Its morphology, leaf anatomy, karyology, palynology, ecology and taxonomic position are examined. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 106–114.     

Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in China

Isoetes yunguiensis is an endangered and endemic fern in China.Field survey indicated that only one population and no more than 50 individuals occur in the wild.The genetic variation of 46 individuals from the population remaining at Pingha (Guizhou Province,China)was assessed by Random Amplified Polymorphic DNA (RAPD)fingerprinting.Twelve primers were screened from sixty ten-bp arbitrary primers,and a total of 95 DNA fragments were scored.Of these,62.1%were polymorphic loci,which indicated that high level genetic variation existed in the natural population.The accumulation of genetic variation in the history of the taxon and the apparent minimal reduction effect on genetic diversity following destruction of habitat might be responsible for the high level genetic diversity presently remaining in the I.yunguiensis population.However,with the continuing decrease of population size,the genetic diversity will gradually be lost.We suggest that the materials from the extant population should be used for re-establishment of the populations.     

Genetic diversity of the endangered Chinese endemic herb Primulina tabacum (Gesneriaceae) revealed by amplified fragment length polymorphism (AFLP)

Primulina tabacum Hance, is a critically endangered perennial endemic to limestone area in South China. Genetic variability within and among four extant populations of this species was assessed using AFLP markers. We expected a low genetic diversity level of this narrowly distributed species, but our results revealed that a high level of genetic diversity remains, both at population level (55.5% of markers polymorphic, H _E = 0.220, I _S = 0.321), and at species level (P = 85.6% of markers polymorphic, H _E = 0.339, I _S = 0.495), probably resulting from its refugial history and/or breeding system. High levels of genetic differentiation among populations was apparent based on Nei’s genetic diversity analysis (G _st=0.350). The restricted gene flow between populations is a potential reason for the high genetic differentiation. The population genetic diversity of P. tabacum revealed here has clear implications for conservation and management. To maintain present levels of genetic diversity, in situ conservation of all populations is necessary.     

RAPD Analysis Reveals Geographic Differentiations within Allium schoenoprasum L. (Alliaceae)

Abstract: Random amplified polymorphic DNA (RAPD) analysis was used to study the phylogenetic relationships between species in Allium section Schoenoprasum and for the investigation of the intraspecific differentiation of A. schoenoprasum. RAPD analysis of 39 samples representing eight species of sect. Schoenoprasum and one sample of A. atrosanguineum (sect. Annuloprasum ) resulted in 233 interpretable RAPD bands. The analysis clearly distinguishes the species of section Schoenoprasum. The arrangement of the accessions of A. schoenoprasum in all dendrograms mirrors the geographical distribution, with a clear differentiation between an Asian and European subgroup. Within the European group, Scandinavian material is clearly distinct from S and E European material. Informally described morphological types of A. schoenoprasum could not be confirmed by RAPD analysis but represent recurrent ecological adaptations. A combination of phenetic (UPGMA, neighbour-joining analysis), cladistic (parsimony analysis), and statistical (PCA) methods of data analysis resulted in clearer phylogenetic interpretations than each of the methods facilitates when used separately.     

Genetic diversity and structure of the narrow endemic Wyethia reticulata and its congener W. bolanderi (Asteraceae) using RAPD and allozyme techniques

Wyethia reticulata is an edaphic endemic in the Sierra Nevada foothills. Its sympatric congener, W. bolanderi, is also restricted to the foothills, but has a north-south range of 275 km, compared to 14 km for W. reticulata. The goals of this study were to determine clonal diversity, population size, genetic variation, and spatial and generic structure for each species from paired populations in El Dorado County, California, using allozyme and RAPD (random amplified polymorphic DNA) methodologies. Wyethia reticulata, spreading by rhizomes, had populations dominated by a few large individuals, while W. bolanderi, with a basal caudex, had populations of a few hundred evenly sized individuals. Genetic analyses indicated that W. reticulata, compared to its congener, had somewhat less genetic diversity (H(T): 0.28 vs. 0.38), had more of its genetic variation partitioned among populations (F(ST): 0.25 vs. 0.07), and showed a complete absence of inbreeding (F(IS): -0.03 vs. 0.22). Population membership in accord with populations defined by geographical location resulted only when all markers were included in the analysis. Ecological limits on recruitment of genets appears to result in small population size in W. reticulata. Limited gene flow, drift within small populations, and sexual reproductive dominance of large clones result in the genetic divergence of populations in this species, while genetic diversity is maintained by the longevity of clones and outbreeding.     

Genetic variability of the narrow endemic tree Antirhea aromatica Castillo-Campos Lorence, (Rubiaceae, Guettardeae) in a tropical forest of Mexico

BACKGROUND AND AIMS: Genetic structure and variability were examined in the only three extant populations of the narrow-endemic tree Antirhea aromatica (Rubiaceae, Guettardeae), an endangered species of the tropical forest of eastern Mexico. Patterns of genetic diversity within and among populations for adult plants and seedlings were obtained. METHODS: Allozyme electrophoresis of 15 loci was conducted and the data analysed with statistical approximation for obtaining genetic diversity, structure and gene flow. KEY RESULTS: The mean expected heterozygosity (He) in the adult and seedling populations was 0.18 +/- 0.08 and 0.20 +/- 0.09, respectively. The genetic variation explained by differences among populations was 51 and 35 %, for adult and seedling populations, respectively. On average, gene flow between paired adult populations was low (Nm = 0.26 +/- 0.09), compared with other trees from the tropical forest. CONCLUSIONS: The results indicated that the populations evaluated have high genetic variability, compared with other endemic and geographically narrowly distributed plant species, in areas with high levels of environmental heterogeneity (e.g. tropical forests). The conservation implications of the results are discussed, and in this regard it is proposed that A. aromatica should be considered as an indicator species with economic potential. It is suggested that sustainable management practices should be implemented and that the areas where the species is distributed should be declared a natural reserve to ensure the species conservation.     

Genetic diversity and structure of the endemic Caesalpinia hintonii complex (Leguminosae: Caesalpinioideae) in Mexico

The Caesalpinia hintonii complex is formed by five endemic species (C. hintonii, C. laxa, C. macvaughii, C. melanadenia and C. epifanioi) occurring in central Mexico. This species complex is under incipient genetic divergence as by-product of local adaptations in reproductive and morphological traits to different habitats. We estimate the genetic variation and structure of populations of this species complex to assess the extent of genetic differentiation among populations and related species along its geographic distribution. Estimations of genetic diversity and structure were done based on ten enzymes and 18 loci. Mean number of alleles per locus ranged from 1.5 to 1.9. Polymorphic loci ranged from 42.1 to 68.4. Observed (H_o: range 0.191–0.275) and expected (H_e: range 0.205–0.317) heterozygosities in this complex were higher compared with other endemic and legume species. Neis genetic diversity estimates showed that most genetic variation was found within (H_S = 0.325) rather than among populations (D_ST = 0.085). Populations of the species C. hintonii showed a considerable genetic differentiation (F_ST = 0.207). The results of genetic diversity and structure within and among populations are in accord with the great morphological differentiation described for this species complex.     

Allozyme Variation and the Genetic Structure of Populations of the Rare Sedge Carex misera (Cyperaceae)

Abstract Carex misera is a rare sedge, endemic to rocky outcrops and mountain summits within the southern Appalachian Mountains from northern Georgia to northern North Carolina. We assessed allozyme diversity for 406 individuals from nine populations over most of the geographic range. Twenty-seven putative loci were assayed and eight (30%) were found to be polymorphic. Nei's gene diversity statistics ( H _T =0.043, H _S =0.019, G _ST =0.551) indicated low levels of variation but relatively highly differentiated populations, suggesting little gene flow. Significant deviations from genotypic expectations under Hardy-Weinberg equilibrium, high positive fixation indices, and the existence of small genetic neighborhoods within populations suggest that at least some inbreeding occurs. Cluster analysis of Nei's genetic identity statistics and principal component analysis of allele frequency data showed high similarity among the six southern populations with the two northern populations more differentiated from them and from each other. These results suggest that preservation of the northern populations is necessary to conserve the already low levels of genetic diversity within the species.     

岛屿特有种全缘冬青遗传多样性的ISSR分析

全缘冬青(Ilex integra)为岛屿特有植物,主要分布于浙江舟山群岛,被列为浙江省珍稀濒危植物。作者运用ISSR分子标记对舟山群岛的6个全缘冬青种群(5个自然种群和1个栽培种群)共57个个体进行了遗传多样性分析。采用9条随机引物扩增出78条清晰谱带,其中45条为多态性条带,多态位点百分比(PPL)为57.7%。经POPGENE软件分析,全缘冬青种群平均多态位点百分比(PPL)为42.1%,Nei’s基因多样度(HE)为0.153,较其他海岛植物的遗传多样性偏低。5个自然种群遗传分化系数Gst=0.316,AMOVA测度Fst=0.295,Shannon多样性分化系数S=0.304,遗传分化程度较高。全缘冬青种群间地理距离与遗传距离具有正相关性(r=0.649,P<0.05),岛屿间地理隔离对种群间遗传分化产生了较为显著的影响。UPGMA聚类分析显示,朱家尖岛与普陀岛种群亲缘关系较近,而舟山岛栽培种群是由桃花岛自然种群移植而来。针对全缘冬青种群遗传多样性较低及遗传分化程度较高的现状,应加强现有自然种群的就地保护,促进种群自然更新;建立种质资源库,收集不同岛屿的种源进行混合繁殖,促进基因交流。     

缙云山特有植物缙云黄芩的遗传多样性研究

采用聚丙烯酰胺凝胶垂直平板电泳检测了重庆缙云山特有植物缙云黄芩7个居群70个个体的过氧化物酶、细胞色素氧化酶、超氧化物歧化酶、淀粉酶和酯酶5种等位酶,获得20个基因位点的资料,由此分析了其遗传分化水平。结果表明：缙云黄芩遗传多样性水平较高,多态位点比率P为45．7％,平均每个多态位点的等位基因数目A为1．46,平均预期杂合度He为0．205,平均观察杂合度Ho为0．352。缙云黄芩居群间遗传分化水平较高,Gst=0．401,居群间遗传一致度和遗传距离的平均值分别为0．741和0．300。缙云黄芩遗传多样性的40．1％来源于居群之间的基因差异,59．9％属于居群内的遗传分化,因而对缙云黄芩遗传多样性的保护,应保护其较多的居群。     

Genetic diversity in Nothofagus alessandrii (Fagaceae), an endangered endemic tree species of the coastal maulino forest of Central Chile

BACKGROUND AND AIMS: The endemic tree Nothofagus alessandrii (Fagaceae) has been historically restricted to the coastal range of Region VII of central Chile, and its forests have been increasingly destroyed and fragmented since the end of the 19th century. In this study, the patterns of within- and among-population genetic diversity in seven fragments of this endangered narrowly endemic tree were examined. METHODS: Allozyme electrophoresis of seven loci of N. alessandrii was used to estimate genetic diversity, genetic structure and gene flow. KEY RESULTS: High levels of genetic diversity were found as shown by mean expected heterozygosity (H(e) = 0.182 +/- 0.034), percentage of polymorphic loci (P(p) = 61.2 %), mean number of alleles per locus (A = 1.8) and mean number of alleles per polymorphic locus (A(p) = 2.3). Genetic differentiation was also high (G(ST) = 0.257 and Nm = 0.7). These values are high compared with more widespread congeneric species. CONCLUSIONS: Despite its endemic status and restricted geographical range N. alessandrii showed high levels of genetic diversity. The observed patterns of diversity are explained in part by historical processes and more recent human fragmentation.     

Multiple autopolyploidizations and range expansion of Allium przewalskianum Regel. (Alliaceae) in the Qinghai‐Tibetan Plateau

We used the Allium przewalskianum diploid–tetraploid complex on the Qinghai‐Tibetan Plateau (QTP) as a model to examine how this complex responded to the Quaternary climatic oscillations, and whether multiple autopolyploidizations have occurred. We sequenced five chloroplast DNA (cpDNA) fragments (accD‐psaI, trnH‐psbA, trnL‐trnF, trnS‐trnG and rpl16‐intron) in 306 individuals (all of known ploidy level) from 48 populations across the distribution of this species complex. We identified a total of 32 haplotypes—11 in diploids only, 13 in tetraploids only, and 8 found in both cytotypes. This, plus network analyses, indicated that tetraploids have arisen independently from diploids at least eight times. Most populations in the eastern QTP contained multiple haplotypes, but only a single haplotype was found for 17 tetraploid populations on the western QTP, suggesting a recent colonization of the western QTP. We further found that this species complex underwent an earlier range expansion around 5–150 thousand years ago (kya), after the largest glacial period (800–170 kya) in the QTP. In addition, the high frequencies of tetraploids in the QTP suggested that the tetraploid A. przewalskianum cytotype has evolutionary advantages over diploids in colonizing and/or surviving the arid habitats of the QTP.