Allozyme variation in the endangered insular endemic Castilleja grisea

BACKGROUND AND AIMS: Genetic diversity in Castilleja grisea, an endangered, perennial herb endemic to San Clemente Island, California was investigated. Subsequent to the elimination of goats from the island in 1992, many populations of C. grisea have reappeared and have been increasing in size. METHODS: Nineteen populations were surveyed for their genotype at 19 allozyme loci. KEY RESULTS: At the taxon level, 57.9 % of loci are polymorphic with A(P) = 3.09 and H(E) = 0.137. Populations averaged 33.0 % polymorphic loci with A(P) = 2.43 and H(E) = 0.099. Most variation is found within rather than among populations (G(ST) = 0.128), although differentiation among populations is significant. Genetic identities range from I = 0.960 to I = 1.000 with mean I = 0.990. There is no significant relationship between genetic and geographic distance. Gene flow among populations is Nm = 2.50 based on private alleles and Nm = 1.70 based on F(ST). Outcrossing rates based on fixation indices average t = 1.01, indicating a primarily out-crossed mating system. CONCLUSIONS: The observed genetic variation is moderately high, unusually so for an insular endemic species, suggesting that C. grisea may not have lost substantial genetic variation during 150 years of overgrazing, and indicating that it is unlikely to be endangered by genetic factors.     

Genetic diversity and differentiation in Camellia reticulata (Theaceae) polyploid complex revealed by ISSR and ploidy

Camellia reticulata is a well-known ornamental and oil plant that is endemic to southwest China. This species shows three cell ploidies, i.e., diploidy, tetraploidy and hexaploidy. We made the first investigation of genetic diversity and differentiation of natural populations of C. reticulata, and 114 individuals from 6 populations were sampled. Cytogeography results showed that ploidy is invariable within populations and evenly distributed. A relatively high level of genetic diversity was found in C. reticulata, both at the species level (PPB = 88.89%; H = 0.2809; I = 0.4278) and at the population level (mean PPB = 42.13%; mean H = 0.14; mean I = 0.21). We found a relatively low degree of differentiation among ploidies (G(ST) = 0.2384; AMOVA = 10.26%) and a relatively high degree of differentiation among populations (G(CS) = 0.3807; AMOVA = 48.75%). The high genetic diversity can be explained by its biological character, wide distribution and ploidies, and the special genetic structure can be ascribed to polyploid origin from hybridization with different Camellia spp. This information will be useful for the introduction, conservation and further studies of C. reticulata and related species.     

Genetic Structure Within and Among Populations of Saruma henryi, an Endangered Plant Endemic to China

The endangered perennial plant Saruma henryi Oliv. is endemic to China and has phylogenetic, ecological, and medicinal value. We used 10 microsatellite (SSR) loci to investigate genetic diversity and differentiation in 16 natural populations. Genetic diversity was high at the species level (H(E) = 0.9427, h = 0.9410, I = 3.0213) and low at the population level (H(E) = 0.4441, h = 0.4307, I = 0.6822). Pronounced genetic differentiation was detected among populations (G(ST) = 0.5428, F(ST) = 0.5524), in line with the limited among-population gene flow (Nm = 0.21). The significant isolation-by-distance pattern revealed by a Mantel test (r = 0.311, P = 0.001) suggested a clear geographic tendency in the distribution of genetic variability. Bayesian assignment and principal coordinates analyses supported the clustering of 16 populations into three groups. The present SSR results were also compared with previously published ISSR results. These results have significant implications for conservation of the species.     

Population genetics and breeding system of Tupistra pingbianensis (Liliaceae),a naturally rare plant endemic to SW China

The levels and partitioning of genetic diversity and inbreeding depression were investigated in Tupistra pingbianensis, a narrow endemic of southeast Yunnan, China, characterized by a naturally fragmented distribution due to extreme specialization on a rare habitat type. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using amplified fragment length polymorphism markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P_(99)=96.012%; H_t=0.302) and at the population level (P_(99)=51.41%; H_8= 0.224). Strong genetic differentiation among populations was also detected (F_(ST)= 0.2961; θ~Ⅱ= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing, and outcrossing plant species. This result was consistent with mating patterns detected by our pollination experiments. The indirect estimate of gene flow based on θ~Ⅱwas low (N_m=0.64). Special habitat and its life history traits might play an important role in shaping the genetic diversity and the genetic structure of this species. A pollination experiment also failed to detect significant inbreeding depression upon F_1 fruit set, seed weight, and germinate rate fitness-traits. As a naturally rare species, T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history, we propose this species need only periodic monitoring to ensure their continued persistence, but not intervention to remain viable.     

Diversity of Arabidobsis thaliana (L.) Heynh. natural populations at the northern limits of the species range: RAPD analysis

In natural populations of Arabidobsis thaliana (L.) Heynh., occupying northern limits of the species range (Karelia), the level of genetic diversity was evaluated. In two insular and one mainland population variability at 82 RAPD loci was tested. Considerable genetic diversity revealed (P = 4.5%; Hexp = 0.177) was not typical of self-pollinating plant species. It was demonstrated that genetic differentiation among the populations (G(ST)= 0.680) was rather high, pointing to the low level of gene flow in the isolated insular populations. It was suggested that the high level of Arabidopsis population polymorphism in Karelia could be associated with extreme growing conditions at the northern limits of the species range.     

ISSR variation in the endemic and endangered plant Cycas guizhouensis (Cycadaceae)

BACKGROUND AND AIMS: Cycas guizhouensis (Cycadaceae) is a rare and endangered species endemic to the southwest of China. An investigation was undertaken into the genetic variation of wild populations. METHODS: ISSR markers were used to determine the genetic variation within and between 12 extant populations of this species. KEY RESULTS: Low genetic diversity (at population level, P = 14.21 %, H(E) = 0.0597; at species level, P = 35.90 %, H(T) = 0.1082) and a high degree of differentiation among populations (G(ST) = 0.4321) were detected. CONCLUSIONS: This genetic structure is considered to be due to the combined effects of slow biochemical evolution, genetic drift, inbreeding and limited gene flow between populations. Based on these findings, strategies are proposed for the genetic conservation and management of the species.     

Differentiation among populations of the Rhodiola iremelica Boriss. (Grassulaceae). in the Southern Urals

Isoenzyme markers and polyacrylamide gel electrophoresis have been used to study the genetic structure of populations of Rhodiola iremelica Boriss. (Grassulaceae), a Southern Ural endemic protected by the state and included in the Red Data Book of Bashkortostan Republic. A relatively large genetic variation at the species level has been found. The subdivision among populations (F(ST) = 0.115) is higher than in most cross-pollination angiosperms. No consistent pattern has been observed in the spatial distribution of its genetic variation. The relatively high differentiation among samples of R. iremelica characterized by small effective population sizes, may be accounted for by genetic drift, inbreeding, and a restricted gene flow. To preserve the population gene pool, in situ protection of the species in nature is insufficient. It seems advisable to create synthetic populations ex situ and reintroduce them into nature.     

Conservation genetics of the endangered Iberian steppe plant Ferula loscosii (Apiaceae)

Ferula loscosii (Lange) Willk (Apiaceae) is a threatened endemic species native to the Iberian Peninsula. The plant has a narrow and disjunct distribution in three regions, NE, C and SE Spain. Genetic variability within and among 11 populations from its natural distribution was assessed using allozymes. Intermediate levels of genetic diversity were detected in F. loscosii (P(99%) = 36.83; H(E) = 0.125; H(T) = 0.152). However, the highest genetic diversity (58%) corresponded to the threatened populations from SE and C Spain (H(T) = 0.169) rather than the more abundant and larger populations from NE Spain (Ebro valley) (H(T) = 0.122). Low to moderate levels of genetic structure were found among regional ranges (G(ST) = 0.134), and several statistical spatial correlation analyses corroborated substantial genetic differentiation among the three main regional ranges. However, no significant genetic differentiation was found among the NE Spain populations, except for a northernmost population that is geographically isolated. Outcrossing mating and other biological traits of the species could account for the maintenance of the present values of genetic diversity within populations. The existence of an ancestral late Tertiary wider distribution of the species in SE and C Spain, followed by the maintenance of different Quaternary refugia in these warmer areas, together with a more recent and rapid post-glacial expansion towards NE Spain, are arguments that could explain the low genetic variability and structure found in the Ebro valley and the higher levels of diversity in the southern Iberian populations.     

Genetic diversity analysis and conservation of the endangered Chinese endemic herb Dendrobium officinale Kimura et Migo (Orchidaceae) based on AFLP

Dendrobium officinale is a critically endangered perennial herb endemic to China. Determining the levels of genetic diversity and patterns of population genetic structure of this species would assist in its conservation and management. Data of 12 populations were used to assess its genetic diversity and population structure, employing the method of amplified fragment length polymorphism (AFLP). A high level of genetic diversity was detected (H (E) = 0.269) with POPGENE. As revealed by AMOVA analysis, there was moderate variation between pairs of populations with Phi(ST) values ranging from 0.047 to 0.578 and on average 26.97% of the genetic variation occurred among populations. Three main clusters were shown in UPGMA dendrogram using TFPGA, which is consistent with the result of principal coordinate ananlysis (PCO) using NTSYS. Keeping a stable environment is critical for the in situ conservation and management of this rare and endangered plant, and for ex situ conservation it is important to design an integrated germplasm bank.     

Genetic structure of Galitzkya macrocarpa and G. potaninii, two closely related endemics of central Asian mountain ranges

BACKGROUND AND AIMS Habitats in mountains are often isolated. Plants growing in these sites face severe dispersal limitations, but also difficulties for recruitment. The focus was laid on the magnitude of genetic differences among populations but also on the size of potentially occurring clones. METHODS: RAPD fingerprints were obtained from 23 populations in southern Mongolia. Sampling covered the entire distribution range of Galitzkya macrocarpa; samples of G. potaninii represented only the Mongolian part of its mainly northern Chinese range. KEY RESULTS: The Mongolian endemic G. macrocarpa showed moderately strong population differentiation (Phi ST = 0.251), and limited evidence for isolation by distance. Local genetic diversity was not positively correlated to habitat size, and not reduced in peripheral populations. Clonal growth is possible, but most plants originate from sexual reproduction. In contrast, populations of G. potaninii were highly differentiated (Phi ST = 0.550); and the most remote outposts had reduced genetic diversity. In these areas, isolation is expected to date back to glacial times. CONCLUSIONS: Effects of natural fragmentation differ among species. Both are rare, but G. macrocarpa appears to be able to maintain genetic diversity over its range. Clonal growth is an option in its mixed reproduction strategy and allows survival under harsh conditions. In contrast, genetic structure in G. potaninii gives reason for concern, and further studies on population dynamics are needed.     

Genetic diversity in harvested and protected populations of wild American ginseng, Panax quinquefolius L. (Araliaceae)

Genetic diversity was examined at 16 allozyme loci in 21 wild populations of the medicinal plant American ginseng, Panax quinquefolius L. (Araliaceae). This species has been harvested from forests in North America for more than 250 years. Average expected heterozygosity was significantly greater within protected populations (H(e) = 0.076) than within populations in which harvesting was permitted (H(e) = 0.070). More notably, genetic structure was greater among unprotected populations (G(ST) = 0.491) than among protected populations (G(ST) = 0.167). These differences in the level and distribution of genetic diversity in American ginseng populations indicate that harvesting may have significant evolutionary implications for this species. Age class structure also shifted toward smaller, nonreproductive plants in unprotected populations. Juvenile plants had lower genetic diversity (H(e) = 0.067) than reproductive plants (H(e) = 0.076) suggesting that conserving a proportion of the largest (oldest) plants in each population is important to protect reproductive fitness and the evolutionary potential of the species. Due to its high genetic structure, conservation recommendations include protecting populations throughout the range of P. quinquefolius.     

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.     

Phylogeography of North African Atlas cedar (Cedrus atlantica, Pinaceae): Combined molecular and fossil data reveal a complex Quaternary history

Northwest Africa is a major hotspot of plant biodiversity, but very little is known about the Quaternary range dynamics of plant species in this region. Here we investigate the range-wide population structure and phylogeography of Atlas cedar (Cedrus atlantica), an emblematic forest tree endemic to Morocco and Algeria. We genotyped 261 individuals from 11 populations using AFLP markers. Data were analyzed using both conventional F(ST)-based techniques and Bayesian clustering. Overall population differentiation was high (F(ST) = 0.25). Two major groups of populations were identified, one distributed through the Rif and Middle Atlas mountains in Morocco and the other through the Algerian Tell Atlas and Aurès mountains as well as the Middle Atlas. Combined molecular and fossil data indicate that C. atlantica survived the Last Glacial Maximum in at least three disjunct refugia along the coast of the Mediterranean Sea, whereas the Middle Atlas, today the core of the species range, has been colonized relatively recently (<10000 yr BP). The colonization history of individual populations has left clear imprints in their present-day diversity, which may vary greatly even between nearby stands. Our study illustrates how integrating different data sources and analytical approaches can help elucidate complex range dynamics that would otherwise remain undeciphered.     

Phylogeography of the montane caddisfly Drusus discolor: evidence for multiple refugia and periglacial survival

We studied the genetic population structure and phylogeography of the montane caddisfly Drusus discolor across its entire range in central and southern Europe. The species is restricted to mountain regions and exhibits an insular distribution across the major mountain ranges. Mitochondrial sequence data (COI) of 254 individuals from the entire species range is analysed to reveal population genetic structure. The data show little molecular variation within populations and regions, but distinct genetic differentiation between mountain ranges. Most populations are significantly differentiated based on F(ST) and exact tests of population differentiation and most haplotypes are unique to a single mountain range. Phylogenetic analyses reveal deep divergence between geographically isolated lineages. Combined, these results suggest that past fragmentation is the prominent process structuring the populations across Europe. We use tests of selective neutrality and mismatch distributions, to study the demographic population history of regions with haplotype overlap. The high level of genetic differentiation between mountain ranges and estimates of demographic history provide evidence for the existence of multiple glacial refugia, including several in central Europe. The study shows that these aquatic organisms reacted differently to Pleistocene cooling than many terrestrial species. They persisted in numerous refugia over multiple glacial cycles, allowing many local endemic clades to form.     

Genetic Variability and Differentiation Among Populations of the Azorean Endemic Gymnosperm Juniperus brevifolia: Baseline Information for a Conservation and Restoration Perspective

The Azorean endemic gymnosperm Juniperus brevifolia (Seub.) Antoine is a top priority species for conservation in Macaronesia, based on its ecological significance in natural plant communities. To evaluate genetic variability and differentiation among J. brevifolia populations from the Azorean archipelago, we studied 15 ISSR and 15 RAPD markers in 178 individuals from 18 populations. The average number of polymorphic bands per population was 65 for both ISSR and RAPD. The majority of genetic variability was found within populations and among populations within islands, and this partitioning of variability was confirmed by AMOVA. The large majority of population pairwise F(ST) values were above 0.3 and below 0.6. The degree of population genetic differentiation in J. brevifolia was relatively high compared with other species, including Juniperus spp. The genetic differentiation among populations suggests that provenance should be considered when formulating augmentation or reintroduction strategies.     

滇牡丹遗传多样性的ISSR分析

应用ISSR标记对中国西南地区特有植物滇牡丹(Paeonia delavayi)的遗传多样性进行了研究。从100个引物中筛选出10个用于正式扩增,在取自16个自然居群和1个迁地保护居群的511个个体中,检测到92个多态位点。在居群水平上,多态位点百分率（PPB）为44.61%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.1657和0.2448。在物种水平上,多态位点百分率（PPB）为79.31%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.2947和0.4355。居群间的遗传分化系数(G_ST)达0.4349。结果表明：滇牡丹遗传多样性水平较高,居群间遗传分化较大。结合以前的研究结果,对滇牡丹的现状进行评估的结果显示,滇牡丹并不濒危。     

High levels of genetic diversity in island populations of the island endemic Suzukia luchuensis (Labiatae)

Genetic diversity and genetic differentiation within and among island populations was examined by allozyme electrophoresis in Suzukia luchuensis (Labiatae), which is endemic to four of the Ryukyu Islands, southern Japan, and one island near Taiwan. Intrapopulation allozyme diversity was very low in all the four Ryukyu Islands, probably due to the effects of random drift in small populations. In contrast, genetic diversity at the species level was high, possibly because of an ancient origin of populations and/or multiple colonization of the species on different islands. Genetic differentiation among the overall populations was high (G(ST)=0.863), while gene flow (Nm) as estimated from allozyme frequency data was 0.041, suggesting that its occurrence among populations is highly restricted. Hierarchical analysis of genetic differentiation indicated that a high proportion of the total allelic variance is attributed to variation among islands, corresponding to the fact that several alleles were fixed on only one island. However, intraisland genetic differentiation was small on all islands except Yonaguni Island, where S. luchuensis is relatively widely distributed. Most diversity was thus due to differences among islands.     

Genetic diversity and population structure of the endangered species Psammosilene tunicoides revealed by DALP analysis

Psammosilene tunicoides W. C. Wu et C.Y. Wu (Caryophyllaceae) is a rare plant endemic to China. We used the direct amplification of length polymorphism (DALP) method to analyse the genetic diversity of 17 natural populations of this species from Yunnan, Sichuan and Guizhou provinces. The percentage of polymorphic bands (PPB) was 99.1% at the species level and 19.9% at the population level. The total genetic diversity (Ht) was 0.2827; the genetic diversity within populations (H_s) was 0.0594 and the coefficient of gene differentiation (G_st) was 0.7899. The results indicated low levels of genetic diversity within populations and extremely high genetic differentiation among populations. This pattern might be caused by genetic drift resulting from intensive harvesting and habitat fragmentation. It might also be a result of local adaptation. We further performed experiments on its breeding system and found a high rate of selfing in P. tunicoides, which may be partially responsible for its population structure.     

Population studies on an endemic troglobitic beetle: geographical patterns of genetic variation, gene flow and genetic structure compared with morphometric data

Highly specialized obligatory cave beetles endemic to the French Pyrenees offer an opportunity to investigate the relative importance of environmental conditions and ecological characteristics on the organization of genetic variability, to describe the genetic structure of populations, and to assess the extent of gene flow between local populations in relation to geologic structure. Twenty-three geographically close populations of the beetle Speonomus hydrophilus occurring both in caves (reduced fluctuations in many abiotic parameters) and under the deepest layer of soil in mountains (more exposed to climatic variations) were studied. Significant genetic differentiation at 17 allozyme loci was found among populations in close proximity, as well as among those from distant parts of range. On a larger scale, genetic differences among populations appear to result from low dispersal rates between populations. The spatial patterning observed suggests that allozyme frequencies are not responding to environmentally controlled selection. Substantial genetic divergence (F(ST) = 0.112) occurred throughout the range, with important variation in levels of genetic variability (H: 0.065-0.184) among populations. A significant level of substructuring has occurred among the populations with four major geographic areas of similarity indicated. The substructuring of the species into regions suggests an influence of paleoclimatic gradient and paleoenvironment on the population's genetic structure. Also, founder effect and reduced gene flow appear to have influenced populations in the southeastern portion of the range.     

栌菊木的等位酶分析及其在生物地理和保护生物学上的意义

对我国西南特有的菊科单种属植物栌菊木10个居群、149个个体、11个酶系统及16个酶位点的水平淀粉凝胶电泳分析表明,栌菊木的遗传多样性水平在特有种中较高,在居群水平上,平均每个位点的等位基因数A=1．1—1．4,多态位点百分数P=6．3％—43．8％,实际杂合度Ho=0．063～0．250,期望杂合度He=0．043～0．194;物种水平上A=1．6,P=37．5％,Ho=0．143,He=0．141。居群间遗传一致度I=0．902—1．000,杂合性基因多样度比率FST为0．2395。栌菊木居群间分化程度较大,云南南盘江流域碧云寺居群遗传多样性较低,明显低于金沙江流域的居群。栌菊木可能是来自冈瓦纳古陆祖先的后裔,可能是古地中海退却以后在金沙江干热河谷分化出来的特有属,并且可能由于湿度等生态因子的限制,其分布区未能进一步扩大,仅在南盘江流域形成零散分布。等位酶分析结果还表明栌菊木遗传多样性总体水平较高,建议对遗传多样性较高的金沙江流域的居群加以保护。