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。     

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 of endangered Emmenopterys henryi Oliv. based on ISSR polymorphism and implications for its conservation

Inter-simple sequence repeat (ISSR) markers were used to determine the genetic variation and genetic differentiation of nine populations of Emmenopterys henryi Oliv., an endangered plant endemic to China. Relatively low genetic diversity was detected at population level (the percentage of polymorphic loci P=22.56%, the number of alleles per locus A=1.183+/-0.045, the effective number of alleles per locus A(E)=1.007+/-0.345, Nei's gene diversity h=0.071+/-0.017, Shannon information index I=0.104+/-0.025). However, the genetic diversity at species level was relatively high (P=56.05%; A=1.561+/-0.498, A(E)=1.325+/-0.371, h=0.191+/-0.199, I=0.287+/-0.284). Analysis of molecular variance showed that most of the ISSR variation (68.03%) in E. henryi occurred among populations. The estimated Nm from F (ST )was 0.235. It indicated that the fragmentation and isolation of populations might result from specific evolutionary history and anthropogenic activity. Consequently, genetic drift might play an important role in determining the genetic structure of E. henryi. Conservation strategies for this endangered species are proposed based on the genetic data.     

RAPD analysis of genetic diversity and population structure of Elymus sibiricus (Poaceae) native to the southeastern Qinghai-Tibet Plateau, China

Genetic diversity of Elymus sibiricus (Poaceae) was examined in eight populations from the southeast Qinghai-Tibet Plateau. We detected 291 RAPD polymorphic loci in 93 samples. The percentage of polymorphic bands (PPB) was 79%. Genetic diversity (H(E)) was 0.264, effective number of alleles (N(E)) was 1.444, Shannon's information index (H(O)) was 0.398, and expected Bayesian heterozygosity (H(B)) was 0.371. At the population level, PPB = 51%, N(E) = 1.306, H(E) = 0.176, I = 0.263, and H(B) = 0.247. A high level of genetic differentiation was detected based on Nei's genetic diversity analysis (G(ST) = 32.0%), Shannon's index analysis (33.7%), and the Bayesian method (θ(B) = 33.5%). The partitioning of molecular variance by AMOVA demonstrated significant genetic differentiation within populations (60%) and among populations (40%). The average number of individuals exchanged between populations per generation (N(m)) was 1.06. The populations were found to share high levels of genetic identity. No significant correlation was found between geographic distance and pairwise genetic distance (r = 0.7539, P = 0.9996). Correlation analysis revealed a significant correlation (r = 0.762) between RAPD H(E) found in this study and ISSR H(E) values from a previous study.     

Genetic differentiation among Hemarthria compressa populations in south China and its genetic relationship with H. japonica

Within and among populations genetic variance of twelve Hemarthria compressa populations and one Hemarthria japonica population from China were analyzed using inter simple sequence repeat (ISSR). Twelve primers amplified a total of 165 genomic DNA fragments across a total of 148 individuals of which 156 were polymorphic (94.55%). 75.76% of the bands were unique to each species, while the average genetic distance (GD) between one population of H. japonica and twelve populations of H. compressa was 0.44, which suggest that there was distinct differentiation between these two species. In H. compressa, twelve primers produced 145 bands across 145 individuals. High genetic diversity was observed at species level. The percentage of polymorphic loci (P) was 86.21% and Shannon's information index of diversity (I) was 0.357. In contrast, there were relatively low levels of genetic diversity within population (P=32.93%, I=0.174). Analysis of molecular variance (AMOVA) showed that a considerable proportion of genetic variation (48.02%) resided among populations. The coefficient of gene differentiation (G(ST)=48.6%) also suggested that there was strong genetic differentiation among H. compressa populations in southern China. An indirect estimate of the number of migrants per generation (N(m)=0.264) indicated that gene flow was low among populations of this species. Relative high clonal diversity was found, and all local genotypes were found.     

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.     

Molecular diversity and population structure of the forage grass Hemarthria compressa (Poaceae) in south China based on SRAP markers

Hemarthria compressa is one of the most important and widely utilized forage crops in south China, owing to its high forage yield and capability of adaptation to hot and humid conditions. We examined the population structure and genetic variation within and among 12 populations of H. compressa in south China using sequence-related amplified polymorphism (SRAP) markers. High genetic diversity was found in these samples [percentage polymorphic bands (PPB) = 82.21%, Shannon's diversity index (I) = 0.352]. However, there was relatively low level of genetic diversity at the population level (PPB = 29.17%, I = 0.155). A high degree of genetic differentiation among populations was detected based on other measures and molecular markers (Nei's genetic diversity analysis: G(ST) = 54.19%; AMOVA analysis: F(ST) = 53.35%). The SRAP markers were found to be more efficient than ISSR markers for evaluating population diversity. Based on these findings, we propose changes in sampling strategies for appraising and utilizing the genetic resources of this species.     

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。     

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.     

Contrasting microsatellite variation between subalpine and western larch, two closely related species with different distribution patterns

Subalpine larch (Larix lyallii Parl.) and western larch (Larix occidentalis Nutt.) represent two closely related species with contrasting abundance and distribution patterns in Western North America. Genetic diversity at seven informative microsatellite loci was determined for 19 populations of subalpine larch and nine populations of western larch. Contrasting genetic diversity and patterns of population differentiation were observed between the two species. The overall within-population genetic diversity parameters were lower in subalpine larch (A = 3.2; A(P) = 3.6; H(E) = 0.418) than in western larch (A(P) = 5.51; H(E) = 0.580), a pattern that is likely related to historical or demographic factors. No evidence of interspecific hybridization was observed. Significantly more population differentiation (theta = 0.15; R(ST) = 0.07), consistent with more restricted gene flow, was observed for subalpine larch as compared to western larch (theta = 0.05; R(ST) = 0.04). Under the assumption of an infinite allele mutation model, 12 of the 19 subalpine larch populations showed signs of deviation from the mutation-drift equilibrium, which suggests Holocene population bottlenecks and fluctuations in effective population size for this species. None of the western larch populations deviated significantly from the mutation-drift equilibrium. For both species, Mantel's test revealed a significant positive relationship between geographical and genetic distances indicative of isolation by distance. A similar geographical structure was detected in both species, suggesting at least two genetically distinct glacial populations in each species. The various implications for gene conservation are discussed.     

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.     

Allozyme diversity and genetic structure of the leafy cactus (Pereskia guamacho [Cactaceae])

We examined levels of genetic variation and genetic structure in the leafy cactus (Pereskia guamacho) in arid and semiarid zones in Venezuela. We surveyed genetic diversity within 17 populations using 19 allozyme loci. Genetic diversity was relatively high at both the species (P(s) = 89%, A(s) = 3.26, AP(s) = 3.53, H(es) = 0.24) and population (P(p) = 63%, A(p) = 1.90, AP(p) = 2.42, H(ep) = 0.20) levels. A significant deficit of heterozygote individuals was detected within populations in the Paraguana Peninsula region (F(IS) = 0.301). Relatively low levels of population differentiation were detected at macrogeographic (G(ST) = 0.112) and regional levels (G(ST) = 0.044 for peninsula region and G(ST) = 0.074 for mainland region), suggesting substantial genetic exchange among populations; however, gene flow in this species seems to be regulated by the distance among populations. Overall, estimates of genetic diversity found in P. guamacho are concordant with the pattern observed for other cacti surveyed, namely high levels of polymorphism and genetic diversity with one common allele and several rare alleles per locus. Differences in gene dispersal systems between this species and other cacti studied were not reflected in the patterns of genetic diversity observed. The concentration of the highest estimates of genetic variation in northwestern Venezuela suggests a potential reservoir of plant genetic diversity within xerophilous ecosystems in northern South America.     

濒危植物翅果油树种群的遗传多样性和遗传分化研究

利用微卫星（SSR）标记对来自山西和陕西两省的7个翅果油树（Elaeagnus mollisDiels）种群进行遗传多样性和遗传结构研究。10对SSR标记共检测到126个位点,其中多态位点114个。在物种水平上,平均多态位点百分率为90.79%,有效等位基因数（Ne）、Nei基因多样性指数（H）和Shannon多样性指数（I）分别为1.6072、0.3166、0.4603;在种群水平上,多态位点百分率为61.99%,有效等位基因数（Ne）、Nei’s基因多样性指数（H）、Shannon多样性指数（I）分别为1.5445、0.2683、0.3815。遗传分化系数GST为0.2074,表明了翅果油树种群的遗传变异主要存在于种群内。基因流Nm为1.9111〉1,说明种群间基因交流可以阻止由于遗传漂变导致的遗传分化。聚类结果表明,翅果油树种群间的遗传距离与地理距离有一定的相关性,经Mantel检验,种群的地理距离与遗传距离之间呈正相关,但未达到显著水平（p〉0.05）。结果表明,遗传多样性水平与物种本身特性和不同干扰生境有关,濒危植物并不一定表现为遗传变异水平的降低。     

The genetic structure of the remnant populations of Centaurea horrida in Sardinia and associated islands

BACKGROUND AND AIMS: The Mediterranean region is of prime importance to biodiversity at a global level, mainly due to the abundance of endemic plant species. However, information about these species is still scarce, especially at the genetic level. In this paper the first assessment is reported of the genetic structure of Centaurea horrida (Asteraceae), an endemic, sea-cliff-dwelling plant from Sardinia. METHODS: The study was conducted on seven populations covering the entire natural range of the species by means of SSR (microsatellite) markers. KEY RESULTS: A considerable amount of genetic variation was found (average H(e) = 0.603-0.854), together with a medium-high differentiation among populations, as estimated both by F(ST) (0.123) and R(ST) (0.158). Both Bayesian analysis and AMOVA were employed to detect genetic structuring in this species. The results suggest that the origins of the current populations of C. horrida lie in two gene pools. CONCLUSIONS: Despite the restricted range, C. horrida displays high levels of genetic diversity, structured in such a way that three management units could be deemed viable for its conservation. The protected status of the species will probably suffice to prevent the impoverishment of its genetic resources.     

Genetic variability and differentiation of Caragana microphylla populations as revealed by RAPD markers

Genetic variability in random amplified polymorphic DNA (RAPD) was studied in 90 individuals of Caragana microphylla, an outcrossing perennial shrub species, from five natural populations sampled in Inner Mongolia steppe of China on a small scale. Nineteen selected primers were used to amplify DNA samples, and totally 225 bands were detected. The percentage of polymorphic bands within populations ranged form 58.22% to 63.56%, with an average of 60% at the population level and 71.11% at the species level, indicating relatively high genetic variations in C. microphylla species. Shannon's information index (I) and Nei's gene diversity (h) showed the similar trend with each other. According to the analysis of Nei's gene diversity, the percentage of genetic variation among populations was 7.13%, indicating a low level of genetic differentiation among populations. There existed a strong gene flow (Nm = 3.26) among populations. Although AMOVA analysis also revealed most variation was within populations (phi(ST) = 4.1%), a significant proportion was observed among populations (P<0.001) in the present study, suggesting genetic differentiation occurred among populations at a certain extent. Based on Mantel's tests and the results of previous studies, the genetic structure pattern of C. microphylla accorded with the isolation-by-distance model on a very large scale, however, on a small scale, the significant genetic differentiation among populations might be enhanced by the micro-environmental divergence among the sampling sites, rather than by geographic factors. Analysis of the genetic variations of C. microphylla populations provided useful information for the adaptive strategy of Caragana species.     

栓皮栎天然群体SSR遗传多样性研究

利用微卫星(SSR)标记对我国4个省内的5个栓皮栎（Quercus variabilis Bl.）天然群体的遗传多样性进行了研究。16对SSR标记揭示了栓皮栎丰富的遗传多样性：等位基因数（A）平均8.4375个,有效等位基因数(Ne)平均为5.9512个,平均期望杂合度(He)0.8059,Nei多样性指数(h)为0.8041。栓皮栎自然分布区中心地带的群体具有较高的遗传多样性,而人为对森林的破坏将降低林木群体的遗传多样性。栓皮栎群体的变异主要来源于群体内,群体间分化较小,遗传分化系数仅为0.0455。此外,栓皮栎群体间的遗传距离与地理距离之间存在显著的正相关。这些遗传信息为栓皮栎遗传多样性的保护和利用提供了一定依据。Abstract: Genetic diversity of five Quercus variabilis natural populations in four provinces of China was studied with microsatellite (SSR) markers. A relatively high level of genetic diversity was detected in Q. variabilis species with 16 polymorphic microsatellite loci. Average number of alleles (A) and effective number of alleles (Ne) were 8.4375 and 5.9512 respectively. The mean expected heterozygosity (He) was 0.8059 and Nei diversity index (h) was 0.8041. Higher diversity was found with the populations from the central range of the species in contrast to those from peripheral areas and human activities might decrease the genetic diversity of populations. The majority of genetic variation occurred within populations, which could be concluded from the low coefficient of genetic differentiation (Fst=0.0455). In addition, significant correlation was found between geographical distance and genetic distance. All these results present a basis to the conservation and utilization of genetic diversity of Quercus variabilis.     

山东实生板栗居群遗传多样性ISSR分析

采用ISSR分子标记技术对山东省内的10个板栗居群共279个个体的遗传多样性水平及居群遗传结构进行了研究。10个引物共检测到116个位点,其中101个位点为多态位点,占87.07%。POPGENE分析结果表明:板栗具有丰富的遗传变异(在物种水平上,He=0.2697,H0=0.3999;在居群水平上,PPL=64.58,He=0.2004,H0=0.3010)。Neis遗传多样性分析和AMOVA分析表明,各居群间产生了一定程度的遗传分化(GST=0.2414,FST=0.2224)。居群间一定程度的遗传分化可能是由于生境破坏和基因流的障碍(Nm=0.8743)引起。UPGMA聚类分析可知,临沭、莒南、郯城和费县4个居群优先聚成一支,而莱阳居群单独聚为一支。     

Genetic diversity and differentiation in Dalbergia sissoo (Fabaceae) as revealed by RAPD

Dalbergia sissoo, a wind-dispersed tropical tree, is one of the most preferred timber tree species of South Asia. Genetic diversity and differentiation among natural populations of D. sissoo were examined for the first time. We found a relatively high level of genetic diversity in D. sissoo, both at the species level (percentage of polymorphic bands = 89.11%; H = 0.2730; I = 0.4180) and the population level (percentage of polymorphic bands = 68.7%; H = 0.239; I = 0.358), along with a relatively low degree of differentiation among populations (GST = 0.1311; AMOVA = 14.69%). Strong gene flow among populations was estimated, N(m) = 3.3125. The Mantel test suggested that genetic distances between populations were weakly correlated with geographic distances (R = 0.3702, P = 0.1236). The high level of genetic diversity, low degree of differentiation, strong gene flow, and weak correlation between genetic and geographic distances can be explained by its biological character and wide-spread planting. This information will be useful for the introduction, conservation and further studies of D. sissoo and related species.     

Selection of high heterozygosity popcorn varieties in Brazil based on SSR markers

We analyzed genetic structure and diversity among eight populations of popcorn, using SSR loci as genetic markers. Our objectives were to select SSR loci that could be used to estimate genetic diversity within popcorn populations, and to analyze the genetic structure of promising populations with high levels of heterozygosity that could be used in breeding programs. Fifty-seven alleles (3.7 alleles per locus) were detected; the highest effective number of alleles (4.21) and the highest gene diversity (0.763) were found for the Umc2226 locus. A very high level of population differentiation was found (F(ST) = 0.3664), with F(ST) for each locus ranging from 0.1029 (Umc1664) to 0.6010 (Umc2350). This analysis allowed us to identify SSR loci with high levels of heterozygosity and heterozygous varieties, which could be selected for production of inbred lines and for developing new cultivars.     

Microsatellite Analysis of Genetic Variation and Population Subdivision for the Black Muntjac, Muntiacus crinifrons

The black muntjac (Muntiacus crinifrons) is a rare deer found only in a restricted region in east China. Recent studies of mitochondrial DNA diversity have shown a markedly low level of nucleotide diversity for the species, and the Suichang population was genetically differentiated from the two other populations, in Huangshan and Tianmushan mountains. In this study, we extended the analysis of genetic diversity and population subdivision for the black muntjac using data from 11 highly polymorphic nuclear DNA microsatellite loci. Contrary to the results based on mtDNA data, the microsatellite loci revealed that the black muntjac retained a rather high nuclear genetic diversity (overall average H (E) = 0.78). Nevertheless, both types of markers supported the idea that the extant black muntjac population is genetically disrupted (overall phi (ST) = 0.16 for mtDNA and overall F (ST) = 0.053 for microsatellite, both P < 0.001). The correlation between genetic differentiation and geographic distance was not significant (Mantel test; P > 0.05), implying that the patterns of genetic differentiation observed in this study might result from recent habitat fragmentation or loss. Based on the results from the mtDNA and nuclear DNA data sets, two management units were defined for the species, Huangshan/Tianmushan and Suichang. We also recommend that a new captive population be established with individuals from the Suichang region as a founder source.