Low genetic diversity and high genetic differentiation in the critically endangered Omphalogramma souliei (Primulaceae): implications for its conservation

Omphalogramma souliei Franch. is an endangered perennial herb only distributed in alpine areas of SW China. ISSR markers were applied to determine the genetic variation and genetic structure of 60 individuals of three populations of O. souliei in NW Yunnan, China. The genetic diversity at the species level is low with P=42.5% (percentage of polymorphic bands) and Hsp=0.1762 (total genetic diversity). However, a high level of genetic differentiation among populations was detected based on different measures (Nei's genetic diversity analysis: Gst=0.6038; AMOVA analysis: Fst=0.6797). Low level of genetic diversity within populations and significant genetic differentiation among populations might be due to the mixed mating system in which xenogamy predominated and autogamy played an assistant role in O. souliei. The genetic drift due to small population size and limited current gene flow also resulted in significant genetic differentiation. The assessment of genetic variation and differentiation of the endangered species provides important information for conservation on a genetic basis. Conservation strategies for this rare endemic species are proposed.     

西鄂尔多斯特有种四合木种群遗传多样性及遗传分化研究

利用垂直板聚丙烯酰胺凝胶电泳技术,对西鄂尔多斯高原特有种四合木（Tetraena mogolica）4个种群遗传多样性和遗传分化进行了初步研究。电泳结果表明,四合木在种和种群水平维持较高的遗传多样性,1多态位点百分率P＝60％,等位基因平均数A＝1.6,平均期望杂合度He=0.245。4个种群之间遗传分化很小,基因分化系数GST只有0.052,明不同于其它濒危物种。四合木种较高的遗传多样性和极低的种群间分化,说明不同的种群可能有共同的起源,随机遗传漂变和近交衰退不是影响遗传多样性的主要过程。     

High genetic diversity vs. low genetic differentiation in Nouelia insignis (Asteraceae), a narrowly distributed and endemic species in China, revealed by ISSR fingerprinting

BACKGROUND AND AIMS: Nouelia insignis Franch., a monotypic genus of the Asteraceae, is an endangered species endemic in Yunnan and Sichuan Provinces of China. Most of the populations are seriously threatened. Some of them are even at the brink of extinction. In this study, the genetic diversity and differentiation between populations of this species were examined in two drainage areas. METHODS: DNA fingerprinting based on inter-simple sequence repeat polymorphisms was employed to detect the genetic variation and population structure in the species. KEY RESULTS: Genetic diversity at species level was high with P=65.05% (percentage of polymorphic loci) and Ht=0.2248 (total genetic diversity). The coefficient of genetic differentiation among populations, Gst, which was estimated by partitioning the total gene diversity, was 0.2529; whereas, the genetic differentiation between populations in the Jinsha and Nanpan drainage areas was unexpectedly low (Gst=0.0702). CONCLUSIONS: Based on the genetic analyses of the DNA fingerprinting, recent habitat fragmentation may not have led to genetic differentiation or the loss of genetic diversity in the rare species. Spatial apportionment of fingerprinting polymorphisms provides a footprint of historical migration across geographical barriers. The high diversity detected in this study holds promise for conservation and restoration efforts to save the endangered species from extinction.     

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.     

武夷山地伯乐树种群遗传多样性的ISSR分析

伯乐树（Bretschneidera sinensis Hemsl.）为我国特有单型科珍稀濒危植物,具重要的科研价值。本研究采用ISSR分子标记对武夷山脉分布的5个伯乐树天然种群和1个移栽种群进行遗传多样性分析。结果表明伯乐树物种水平遗传多样性较高（PPB：75.70%;H_ES：0.304 5;H：0.450 1）,种群水平则较低（PPB：60.13%;H_EP：0.238 1;H：0.347 5）,MJY种群和BSZ种群分别是所有种群中遗传多样性最高和最低的。5个地理种群间遗传分化程度较高（G_st=0.218 1）,原因可能源于伯乐树的繁殖方式及生境片段化,Mantel检验也证实了地理距离与遗传距离具有显著相关性（r=0.626 7,P＜0.05）。针对伯乐树种群遗传多样性现状,建议加强现有自然种群的就地保护,促进种群自然更新。     

Allozyme variation and population genetic structure of common wild rice Oryza rufipogon Griff. in China

In order to determine the genetic diversity and genetic structure of populations in common wild rice Oryza rufipogon, an endangered species, allozyme diversity was analyzed using 22 loci in 607 individuals of 21 natural populations from the Guangxi, Guangdong, Hainan, Yunnan, Hunan, Jiangxi and Fujian provinces in China. The populations studied showed a moderate allozyme variability (A=1.33, P=22.7%, Ho=0.033 and He=0.068), which was relatively high for the genus Oryza. The levels of genetic diversity for Guangxi and Guangdong were significantly higher than those for the other regions, and thus South China appeared to be the center of genetic diversity of O. rufipogon in China. A moderate genetic differentiation (F_ST=0.310, I=0.964) was found among the populations studied. Interestingly, the pattern of population differentiation does not correspond to geographic distance. An estimate of the outcrossing rate (t=0.324) suggests that the species has a typical mixed-mating system. The deficit of heterozygotes (F=0.511) indicates that some inbreeding may have taken place in outcrossing asexual populations because of intra-clone outcrossing events and ”isolation by distance” as a result of human disturbance. In order to predict the long-term genetic survival of fragmented populations, further studies on gene flow among the remaining populations and the genetic effects of fragmentation are proposed. Finally, some implications for the conservation of endangered species are suggested. Received: 22 June 1999 / Accepted: 20 December 1999     

岛屿植物舟山新木姜子居群遗传多样性的RAPD分析

基于随机扩增多态 DNA(RAPD)方法分析了舟山群岛濒危植物舟山新木姜子 (N eolitsea sericea) 6个居群的遗传多样性及分化程度。 10条随机引物扩增出 84个可分析位点 ,多态位点百分比 (PPL)为 3 8.10 %。经 POPOGENE分析发现 ,舟山新木姜子居群平均水平的多态位点百分比 (PPL )为 2 3 .18% ,Nei' s基因多样度 (HE)为 0 .0 793 ,Shannon信息指数 (H )为 0 .12 0 1,与其它岛屿植物比较具有中等偏低水平的遗传多样性 ;岛屿各居群间遗传分化程度较高 (Gst=0 .3 646) ;地理距离与遗传距离之间具有显著相关性 (r=0 .7697,P=96.62 % ) ,岛屿隔离效应是导致居群间遗传分化的重要因素。结合居群遗传多样性及UPGMA聚类分析 ,推测普陀山岛舟山新木姜子部分个体可能为大猫岛迁入的后裔 ,而朱家尖岛舟山新木姜子则由人为移植自普陀山岛。基于舟山新木姜子的物种保护及资源利用 ,建议加强现有自然居群的就地保护 ,促进居群自然更新 ;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 ;选育优良品系用于海岛植被恢复及园林观赏     

Genetic diversity of wild Cymbidium goeringii (Orchidaceae) populations from Hubei based on Inter-simple sequence repeats analysis

Cymbidium goeringii is a diploid and nonrewarding,bumblebee-pollinated species,which is distributed in China,Japan and Korea Peninsula.This species is now highly endangered due to the mass collection and forest clearance in China.In the present study,we investigated the distribution of genetic variation within and between eleven populations of Cymbidium goeringii in central China by using Inter-simple sequence repeats (ISSR) markers.Eleven primers produced a total of 127 clear and reproducible bands of which 112 were polymorphic.High genetic diversity was detected in Cymbidium goeringii for both population level (P = 63.1%;He = 0.194 5) and species level (P = 88.2%;He = 0.262 8).A higher level of genetic differentiation was detected among populations (GST = 0.244 0,FST = 0.220 7)with Nei's Gsr analysis and analysis of molecular variance (AMOVA),and no correlation was found between geographical and genetic distance.Genetic drift rather than gene flow played an important role in forming the present population structure of Cymbidium goeringii.Limited gene flow among populations and gene drift increase the extinction risk of local populations.Some conservation concerns are therefore discussed together with possible strategies for implementing in situ and ex situ conservation.     

云南金钱槭形态变异与遗传变异的相关性研究

对我国特有珍稀濒危保护植物云南金钱槭的形态变异水平、遗传变异模式以及两者之间的相关性进行了研究。形态学性状分析结果表明 :各居群形态性状变异系数的平均值从大到小排列为 :文山居群 (WSh)、屏边居群 (PB)、黑龙潭居群 (HL T)、蒙自居群 (MZ) ;文山居群与屏边、黑龙潭、蒙自居群间已产生显著或极显著水平的形态差异 ,而后三者间的差异未达显著水平。RAPD分析检测到 10 3个位点 ,其中多态位点 84个 ,云南金钱槭物种水平的多态位点比率为 81.5 5 % ,与其它珍稀濒危植物相比该种的遗传多样性水平不低。 AMOVA和 N ei基因多样性指数分析显示 ,尽管大部分遗传变异仍存在于居群内 (分别为5 7.86 %、5 7.33% ) ,但居群间的遗传变异已达较高水平 (分别为 4 2 .14 %、4 2 .6 7% )。相关分析结果显示 ,云南金钱槭的形态变异与海拔、土壤有机质等生态因子有着显著或极显著水平的相关性 ,但与遗传变异的相关性未达显著水平 ,说明云南金钱槭的形态变异虽然具有一定的遗传基础 ,但可塑性及环境压力对形态变异的产生作用更大一些。基于形态性状和 RAPD数据的聚类分析则进一步说明 ,云南金钱槭的形态变异受到环境因子的强烈影响而与遗传背景的关系不显著     

Low genetic structure in an epiphytic Orchidaceae (Oncidium hookeri) in the Atlantic rainforest of South-eastern Brazil

BACKGROUND AND AIMS: Oncidium hookeri is a neotropical species of epiphytic Orchidaceae found in the Brazilian Atlantic rainforest at the top of the Mantiqueira Range of mountains. The genetic variation of O. hookeri was studied to assess the distribution of genetic variability within and among six populations localized in Atlantic rainforest remnants. Gene flow among populations and the occurrence of recent bottlenecks were investigated in order to infer the degree of isolation of these populations. METHODS: Thirteen polymorphic loci were used for allozyme electrophoresis. The data were analysed by means of standard statistical approaches, to estimate gene diversity and the genetic structure of the populations. KEY RESULTS: The mean gene diversity and allelic richness were H(e) = 0.099 and A = 1.75, respectively. F-statistics revealed high heterozygote deficiencies in all populations (F(IS) = 0.43-0.82). Several rare alleles were found in all the populations, and three populations presented private alleles. Low genetic differentiation among O. hookeri populations was detected (F(ST) = 0.029); natural selection may be involved in PGM locus differentiation among populations. The genetic differentiation between paired populations was low, bearing no correlation with geographic distance (Mantel test: r = -0.34, P = 0.72). Only two populations showed signs of recent bottlenecks. CONCLUSIONS: The heterozygote deficiency found seems to be caused by pollinator behaviour; the low frequencies of several alleles of different loci can be maintained due to clonal propagation. Despite the stochastic nature of the wind-dispersal of seeds to long distances, this process may promote an effective gene flow among populations, thus avoiding genetic differentiation.     

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。     

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。     

华木莲居群遗传结构与保护单元

华木莲(Sinomanglietia glauca)仅分布于江西宜春和湖南永顺, 是我国一级重点保护植物。前人采用RAPD、ISSR以及叶绿体SSCP(single-stranded conformation polymorphism)标记对华木莲进行了居群遗传学研究, 但未包括后发现的湖南居群或未检出居群内遗传变异。为了全面检测华木莲遗传多样性及其空间分布格局, 并据此确定保护单元, 本研究采用细胞核微卫星标记对华木莲所有4个居群共77个个体进行了居群遗传学分析。结果表明, 华木莲具有较低的遗传多样性(平均等位基因数A = 2.604, 平均期望杂合度H_E = 0.423)和较高的遗传分化(F_ST = 0.425)。STRUCTURE和主成分分析(Principal Coordinated Analysis, PCA)将4个居群首先分为江西、湖南两组, 江西的2个居群实际上是同一个自然繁育居群, 而湖南的2个居群则为2个分化明显的自然繁育居群。研究还发现湖南居群存在明显的杂合子过剩现象, 可能是小居群内随机因素造成的。研究结果表明华木莲可能在近期历史上遭受过强烈的瓶颈效应, 导致种群缩小、遗传多样性丧失和居群分化加剧, 需要加强对其进化潜力的保护。在制定保护措施时, 需要考虑其较高的遗传分化水平, 根据遗传结构可以将其划分为3个保护单元, 即湖南居群和江西居群分别为2个进化显著单元, 湖南居群进一步划分为2个管理单元(分别为朗溪乡云盘村和小溪乡鲁家村居群)。     

Analysis of genetic variability of South American wild rice populations (Oryza glumaepatula) with isozymes and RAPD markers

The knowledge of population structure and genetic diversity of wild relatives of rice is needed to investigate their evolutionary history and potential use in breeding programs. Very little is known about the wild rice species ( Oryza spp.), particularly those that are native to South America. A study using isozyme and RAPD markers was conducted to estimate the level of genetic diversity of four South American wild rice populations ( Oryza glumaepatula ) recently collected in the Amazon forest and western Brazil rivers. F -statistics and genetic diversity parameters calculated from isozyme and RAPD markers indicated high values for inbreeding coefficients and differentiation among the four populations. In agreement with this, a pattern of greater variation between than within populations was observed with both types of markers. These findings were corroborated by an AMOVA analysis, which indicated that a large portion of the total genetic variation was attributed to regional divergence. The partition of the AMOVA analysis among populations showed that most of the genetic diversity was due to differences among populations. This distribution pattern of genetic variation of O. glumaepatula populations is in agreement with the expectation for an autogamous species and provides important baseline data for conservation and collection strategies for this species.     

Spatial and temporal population genetic variation and structure of Nothotsuga longibracteata (Pinaceae), a relic conifer species endemic to subtropical China

Nothotsuga longibracteata, a relic and endangered conifer species endemic to subtropical China, was studied for examining the spatial-temporal population genetic variation and structure to understand the historical biogeographical processes underlying the present geographical distribution. Ten populations were sampled over the entire natural range of the species for spatial analysis, while three key populations with large population sizes and varied age structure were selected for temporal analyses using both nuclear microsatellites (nSSR) and chloroplast microsatellites (cpSSR). A recent bottleneck was detected in the natural populations of N. longibracteata. The spatial genetic analysis showed significant population genetic differentiation across its total geographical range. Notwithstanding, the temporal genetic analysis revealed that the level of genetic diversity between different age class subpopulations remained constant over time. Eleven refugia of the Last Glacial Maximum were identified, which deserve particular attention for conservation management.     

中华水韭遗传多样性的RAPD分析

采用RAPD方法对珍稀濒危植物中华水韭(Isoetes sinensis)4个自然居群的48个样品进行了DNA多态性分析。从60个随机引物中筛选出14个有效引物,共产生124条DNA片段,其中72条为多态性条带,总的多态位点百分率(PPB)为58．06％。各居群间多态位点百分率差异显著(0．81％-12．90％)。AMOVA分析结果表明,4个居群间基因分化系数Фst=0．5894,即遗传变异中有相当一部分来源于群体间(58．94％)。日益缩小的种群规模而导致的居群内近交和遗传漂变的发生以及居群间有限的基因交流可能是中华水韭目前遗传结构的主要成因。鉴于目前中华水韭居群内个体数偏少、遗传多样性较低的现状,建议对其进行就地保护并保护尽可能多的生境,对不同自然居群内的个体进行植株相互移栽和育苗移栽,以提高不同居群间的基因交流,尽可能地保护中华水韭的遗传多样性。     

Genetic diversity and genetic relationship of Caragana microphylla, Caragana davazamcii and Caragana korshinskii on the Inner Mongolia Plateau, China

C. microphylla, C. davazamcii and C. korshinskii exhibit a geographical replacement series from east to west on the Inner Mongolia Plateau. Currently, there is still a debate about the taxonomic and genetic relationship among these 3 species. We studied the genetic diversity and genetic relationship among these 3 species by analyzing DNA samples of individual plants from within 10 populations with random amplified polymorphic DNA (RAPD) markers. We identified 678 RAPD loci in total, of which all were polymorphic (PPB = 100%). There were 41 unique loci (6.05%). In general, a trend presented that the genetic diversity of these species decreased from east to west. Further, the genetic diversity was significantly negatively correlated with the local annual mean temperature. Analysis of molecular variation (AMOVA) showed that the genetic variation among these 3 species was only 6.08% of the total genetic variation. Between the species, the genetic variation was insignificant (P = 0.9961). The proportion of genetic variation among populations within each species was 11.90% (P < 0.001) of the total genetic variation, and the total genetic variation mainly existed within the populations (82.02%). Estimated with Shannon's index, genetic differentiation within the populations (Hpop/Hsp) was 0.8013, the coefficient of gene differentiation (Gst) was 0.1603, and the gene flow index (Nm) was 2.6192. This, thus, indicates that there is relatively high gene flow among these populations, and that these 3 species are crossbreeding. The genetic diversity level and the population distribution pattern showed geographic continuity to some extent.     

基于微卫星标记的中国东北地区灰飞虱遗传变异及种群遗传结构分析

【目的】本研究旨在明确我国东北地区灰飞虱Laodelphax striatellus种群遗传变异及种群遗传结构,阐明种群间遗传分化与基因流。【方法】利用9对微卫星引物对采自我国东北地区15个地理种群的375头灰飞虱样品进行测序与分析;利用GeneAlex6.51, GENEPOP4.0.9和STRUCTURE 2.3.4等软件分析灰飞虱地理种群间的遗传多样性、遗传分化、基因流及种群遗传结构。【结果】在所分析的375头灰飞虱个体中,各位点平均有效等位基因数Na=6.898;总体上,灰飞虱不同地理种群遗传多样性较高(平均观测杂合度Ho=0.548;平均期望杂合度He=0.582),各种群间基因流较低(Nm=0.660)。UPGMA聚类树、PCoA及STRCTURE分析结果表明,东北地区灰飞虱种群分为两分支:吉林(JL)和沈阳(SY2012,SY2013和SY2014)种群聚为一支;其余种群聚为一支。AMOVA分析结果表明,灰飞虱遗传变异主要来自种群内(87%),种群间变异水平较低(13%)。【结论】中国东北地区灰飞虱遗传多样性较高,不同地理种群存在一定程度的遗传分化,且基因交流较低,存在一...     

Population genetic structure of Monimopetalum chinense (Celastraceae), an endangered endemic species of eastern China

BACKGROUND AND AIMS: Monimopetalum chinense (Celastraceae) standing for the monotypic genus is endemic to eastern China. Its conservation status is vulnerable as most populations are small and isolated. Monimopetalum chinense is capable of reproducing both sexually and asexually. The aim of this study was to understand the genetic structure of M. chinense and to suggest conservation strategies. METHODS: One hundred and ninety individuals from ten populations sampled from the entire distribution area of M. chinense were investigated by using inter-simple sequence repeats (ISSR). KEY RESULTS: A total of 110 different ISSR bands were generated using ten primers. Low levels of genetic variation were revealed both at the species level (Isp=0.183) and at the population level (Ipop=0.083). High clonal diversity (D = 0.997) was found, and strong genetic differentiation among populations was detected (49.06 %). CONCLUSIONS: Small population size, possible inbreeding, limited gene flow due to short distances of seed dispersal, fragmentation of the once continuous range and subsequent genetic drift, may have contributed to shaping the population genetic structure of the species.     

Conservation of rare species with island-like distributions: A case study of Lasthenia conjugens (Asteraceae) using population genetic structure and the distribution of rare markers

Californian vernal pools, a patchy, island-like habitat, are endangered as a result of habitat destruction. Conservation of the remaining vernal pool habitat is essential for the persistence of several endangered species. We present the first study examining DNA-level genetic diversity within and among populations of a vernal pool plant species. We investigated genetic variation across eight populations of the US federally endangered vernal pool endemic Lasthenia conjugens (Asteraceae) using intersimple sequence repeat (ISSR) markers. Genetic diversity within the species was high (Nei's gene diversity estimate was 0.37), with moderate differentiation among populations (Bayesian F _ST analog of 0.124). Using an amova analysis, we found that the majority of the genetic variation (84%) was distributed within populations. There is a significant relationship between geographical distance and pairwise genetic differentiation as measured by the Bayesian estimate θ^B. The alternative hypotheses of historic geological processes within the Central Valley and contemporary gene flow are discussed as explanations of the data. Because of the vulnerability of the populations, we calculated a probability of loss for rare alleles (fragments) in the populations. Calculations show that sampling only one of the eight populations for ex-situ conservation or restoration will capture approximately 54% of the sampled rare fragments. We believe that one of the sampled populations has become extinct since it was sampled. When removing this population from the above-mentioned calculations, sampling one population will capture only 41.3% of the sampled rare fragments. We recommend sampling strategies for future conservation and restoration efforts of L. conjugens.