东俄洛橐吾遗传变异与分化的ISSR分析

应用ISSR标记对东俄洛橐吾（Ligularia tongolensis）的遗传多样性进行了研究。从100个引物中筛选出8个用于正式扩增。在所研究的8个居群共150个个体中检测到148个多态位点。在居群水平上,多态位点百分率（PPB）为50.45%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.1595和0.2440。在物种水平上,多态位点百分率（PPB）为88.10%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.2811和0.4279。居群间的遗传分化系数（Gst）达0.4355。研究结果表明东俄洛橐吾的遗传多样性水平很高,居群间遗传分化较大。这与其多样化的生态环境是有必然联系的。因适应其多样化的生态环境而形成了遗传多样性;且因其生态环境的不连续性阻碍了居群间的基因交流而产生了遗传分化,即东俄洛橐吾高水平的遗传多样性和遗传分化是适应其分布区多样化生态环境的结果。     

濒危植物连香树居群的遗传多样性和遗传分化研究

利用ISSR分子标记技术对濒危植物连香树10个居群的遗传多样性和遗传变异进行了分析,结果表明:连香树物种水平遗传多样性较高,多态位点百分率(PPB)达到69.59%,Nei’s基因多样性指数(H)和Shannon信息指数(I)分别为0.231 3和0.351 4;而在居群水平上,多态位点百分率(PPB)为30.61%,Nei’s基因多样性指数(H)和Shannon信息指数(I)分别为0.115 6和0.173 3。遗传变异分析表明,居群间遗传分化程度高,遗传分化系数(GST)为0.500 3,居群间基因流Nm为0.527 3。Mantel检测,居群间的遗传距离和地理距离之间不存在显著的相关性。生境的片断化使居群间的基因流受阻,可能是导致居群间高遗传分化和居群水平低遗传多样性的主要原因。     

西双版纳地区流苏石斛遗传多样性的ISSR分析

采用ISSR分子标记技术,对西双版纳分布的兰科濒危植物流苏石斛（Dendrobium fimbriatum）5个居群共114个个体的遗传多样性进行了研究。从100条引物中筛选出了12条用于扩增,共检测到117个位点,其中105个为多态位点。分析结果表明,流苏石斛居群水平遗传多样性较低。在物种水平上,流苏石斛多态位点百分率PPB为89．74％,Nei’s基因多样性指数日为0．3227,Shannon’s多样性信息指数见。为0．4779;在居群水平上,各个居群的多态位点百分率PPB差异较大（6．84％～39．32％）,平均值为23．93％,Nei’s基因多样性指数H为0．0871,各个居群的Shannon’s多样性信息指数见平均为0．1290。AMOVA分析的结果显示,流苏石斛的遗传变异大多数存在于居群间,占总遗传变异的74．79％。基于Nei’s遗传多样性分析得出的居群间遗传分化系数Gst=0．7443。各居群间的Nei’s遗传一致度（I）范围为0．5882～0．8331。Mantel检测发现,居群间的遗传距离和地理距离之间无显著的正相关关系（r=0．2419,P=0．2416）。鉴于流苏石斛的遗传多样性现状和居群遗传结构,我们建议对流苏石斛居群所有个体实施及时的就地保护,同时建立迁地保护居群,促进基因交流。     

西双版纳地区流苏石斛遗传多样性的ISSR 分析

采用ISSR 分子标记技术, 对西双版纳分布的兰科濒危植物流苏石斛( Dendrobium fimbriatum) 5 个居群共114 个个体的遗传多样性进行了研究。从100 条引物中筛选出了12 条用于扩增, 共检测到117 个位点, 其中105 个为多态位点。分析结果表明, 流苏石斛居群水平遗传多样性较低。在物种水平上, 流苏石斛多态位点百分率PPB 为89 .74% , Nei′s 基因多样性指数H 为0 . 3227 , Shannon′s 多样性信息指数Hsp 为0 . 4779 ; 在居群水平上, 各个居群的多态位点百分率PPB 差异较大( 6.84% ～ 39.32% ) , 平均值为23.93% , Nei′s 基因多样性指数H 为0 . 0871 , 各个居群的Shannon′s 多样性信息指数Ho 平均为0.1290。AMOVA 分析的结果显示, 流苏石斛的遗传变异大多数存在于居群间, 占总遗传变异的74 . 79%。基于Nei′s遗传多样性分析得出的居群间遗传分化系数Gst = 0 . 7443。各居群间的Nei′s 遗传一致度( I) 范围为0 . 5882～0 . 8331。Mantel 检测发现, 居群间的遗传距离和地理距离之间无显著的正相关关系( r= 0.2419, P=0.2416) 。鉴于流苏石斛的遗传多样性现状和居群遗传结构, 我们建议对流苏石斛居群所有个体实施及时的就地保护, 同时建立迁地保护居群, 促进基因交流。     

珍稀濒危植物太行菊遗传多样性的ISSR分析

利用ISSR分子标记技术对太行山特有濒危物种太行菊11个自然居群的遗传多样性进行研究。用10个引物对11个居群的122个样品进行扩增,共得到150个扩增位点,其中多态性位点149个,多态位点百分率(PPL)为99.33%。POPGENE分析显示,太行菊具有较高的遗传多样性(H=0.2149,I=0.3455)。沁阳市大西天居群的遗传多样性水平最高(H=0.1910,I=0.2969),山西陵川县大双村居群的遗传多样性水平最低(H=0.1356,I=0.2155)。Nei’s遗传多样性分析表明,11个自然居群间出现了较高的遗传分化(基因分化系数Gst=0.2566,基因流Nm=1.4488)。生境的的片段化和基因流障碍可能是导致太行菊居群间遗传分化显著的主要原因。通过对太行菊居群遗传多样性和遗传结构的分析,该文提出了一些保护策略。     

两广地区家蚕白僵菌的SSRs遗传多样性

为查明广东和广西地区家蚕Bombyx mori病原白僵菌的来源及其菌株间的相互关系, 本研究利用了微卫星标记（simple sequence repeats, SSRs）技术, 分别对采自广东、 广西蚕区的白僵菌菌株居群之间和居群之内的遗传多态性进行了研究。结果发现, 两广白僵菌居群之间的基因分化系数（Gst）是0.0590, 多态位点百分率（PPL）为97.73%, Nei氏基因多样性指数（H）为0.1896, Shannon氏信息多样性指数（I）为0.3165, 表明两广家蚕来源的白僵菌居群间的遗传分化较小; 两个居群内部的遗传多态性研究结果分别是广东白僵菌居群的PPL=68.18%, H=0.1910, I=0.3044, 而广西白僵菌居群的PPL=65.91%, H=0.1713, I=0.1791, 表明广东白僵菌居群的遗传多样性水平较高, 广西白僵菌居群遗传多样性水平相对较低。最后, 利用Nei氏遗传距离进行了两广地区白僵菌菌株间地理来源关系的聚类分析, 结果表明实验室保存菌株单独聚为类群Ⅰ, 而不同采集地的菌株聚为类群Ⅱ。结果反映了生产来源的白僵菌菌株存在遗传多态性和基因分化现象, 暗示了家蚕白僵病病原来源的复杂性, 还说明应用SSRs技术进行家蚕白僵病病原的溯源是一条可行的途径。     

滇东南硬叶兜兰核心种质区群体遗传结构

本研究选取国内主要种质采集区-滇东南石灰岩地区7个不同干扰居群为研究对象,旨在对其居群内和居群间遗传变异进行比较研究,以期对其保护措施的提出提供理论依据。通过利用SRAP标记对167个体的遗传多样性和遗传结构研究,结果表明：10对SRAP引物共扩增出288个位点,多态位点比率（PPB）达81.25%,香侬指数（I）为0.3709,在种水平上的具有较高遗传多样性;而居群水平上的多态位点比率仅为47.92%, 香侬指数为0.2348, 居群间平均Nei’s遗传距离为0.1268。经分子遗传变异方差分析（AMOVA）表明,有66.27%的遗传变异来源于居群内,居群间变异占总变异33.73%,此结果与遗传分化系数（Gst=0.3568）结果吻合,居群间基因流（Nm）为0.902, 不同地区间硬叶兜兰居群存在较高的遗传分化; 7个居群的UPGMA聚类在遗传相似性系数达0.863,聚为两支;经Mantel检测（r =0.298, P＞0.05）,表明居群间遗传距离与地理距离无显著相关性。居群当前较高的遗传分化与其交配系统有关,其次,外在因素：人为采集、生境破坏和片断化造成居群内遗传多样性的丧失,加剧居群间的遗传分化,再次,遗传漂变也是另一重要影响因素;此外,适应性进化亦可能加剧了居群间的遗传分化,而基因流对遗传分化的影响不大。     

滇牡丹天然居群的遗传多样性分析

以云南中部及西北部的6个滇牡丹(Paeonia delavayi)天然居群为研究对象,进行株高、新枝长等9个表型性状的表型多样性分析和ISSR分析。结果表明:9个表型性状变异幅度为0.9%~39.8%,平均值达到了18.9%;居群间生殖器官的变异较大,居群内营养器官更容易产生变异。利用居群间欧式距离进行聚类分析,6个居群聚为4个类群,没有与实际地理位置相吻合,说明表型特征的性状与地理距离的相关性不大。遗传多样性分析结果表明:利用筛选得到的10条引物,在取自6个自然居群、180个个体中,检测到56个多态位点。在居群水平上,多态位点百分率(PPB)为60.2%,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.281和0.414。在物种水平上,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.409和0.596。居群间的遗传分化系数(Gst)达0.319。结果显示,表型性状在居群间和居群内均存在广泛变异。滇牡丹遗传多样性水平较高,居群间遗传分化较大,滇牡丹并不濒危。     

古尔班通古特沙漠荒漠肉苁蓉遗传多样性分析

为了全面了解古尔班通古特沙漠荒漠肉苁蓉居群分布的遗传多样性特点,本研究通过ISSR分子标记技术,利用Nei和Shannon等多样性指数对古尔班通古特沙漠中5个居群166个个体的荒漠肉苁蓉遗传多样性、荒漠肉苁蓉种群和种内的遗传多样性进行分析。在供试材料中,8个引物共扩增出144个多态位点,多态位点百分率达100%,5个居群的多态位点百分率差异在46.53%～77.78%之间。在物种水平上,Nei基因多样度(h)为0.260 4,Shannon多样性指数(I)是0.411 0。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0.222 2,居群间的基因流Nm为1.750 7。研究显示古尔班通古特沙漠中荒漠肉苁蓉多态位点比例高,各居群基因交流较多,不同居群间遗传变异并不明显,这些对肉从蓉资源有效地保护和利用具有重要意义。     

云南泸定百合遗传多样性的表型与ISSR分析

用表型变异分析并结合ISSR分子标记对云南境内10个泸定百合(Lilium sargenttiae)居群进行遗传多样性分析。结果显示:(1)云南10个泸定百合居群的7个表型性状的居群间F值在3.26～19.1之间,表型的居群间差异均达到显著或极显著水平;平均表型分化系数为71.22%,居群间变异(59.92%)大于居群内变异(23.42%),说明居群间表型变异是泸定百合居群变异的主要来源。(2)13个ISSR引物共检测到248个多态位点,物种水平上多态位点率98.80%,Nei’s多样性指数和Shannon多样性指数分别为0.265 5和0.413 1;居群内基因多样度(Hs)为0.175 7,居群间基因分化系数(Gst)0.336 7,Mantel检验显示泸定百合居群在地理距离和遗传距离间具有显著相关性(r=0.804 4,P=0.009 9)。研究表明,云南泸定百合的居群间表型和分子水平均具有较高的遗传多样性,居群间的遗传分化较大,并且分化趋势具有明显的地域性。因此,可选择迁地种植对泸定百合进行有效保护。     

An analysis of genetic diversity in Marphysa sanguinea from different geographic populations using ISSR polymorphisms

The genetic diversity of five wild populations of marine polychaete Marphysa sanguinea found in China was investigated using Inter-Simple Sequence Repeat-PCR (ISSR-PCR) polymorphisms. The results of the ISSR-PCR showed that 108 (90.8%) of the 119 ISSR loci tested were polymorphic. The Shannon's information index value was 0.4981, Nei's gene diversity was 0.3418, and the coefficient of gene differentiation (Gst) was 0.3671, which indicated that the among-population component accounted for 36.7% of the total variation, while the within-population component accounted for 63.3%. A UPGMA tree showed that the five populations clustered into two branches. Populations from Dalian, Xingcheng, and Rushan clustered together, while the two Guangxi populations, A and B, clustered into a unique group. The results indicated that the genetic diversity among the five populations of M. sanguinea is high, which will provide useful information for the protection of biodiversity among marine polychaetes.     

珍稀濒危植物长叶红砂种群遗传多样性的ISSR分析

采用ISSR分子标记技术,对濒危小灌木长叶红砂(Reaumuria trigyna) 集中分布的5个种群的遗传多样性水平和遗传结构进行了研究。14条引物共检测到114个位点,其中99个为多态位点,多态位点比率为86.84％,长叶红砂种群具有较高的遗传多样性。物种水平上Shannon多样性指数(I)为0.468 8,Nei基因多样性指数(H)为0.308 4;种群水平上,多态位点比率P为77.89%,I为0.410 6,H为0.260 9,基因分化系数Gst为0.106 9,揭示了长叶红砂种群遗传变异多存在于种群内,种群间的遗传分化较小,占10.69%。 基因流(Nm)为4.178 7＞1,说明种群间的基因交流,防止了由于遗传漂变导致的遗传分化。聚类分析表明长叶红砂种群遗传距离与地理距离之间无显著的相关性。研究结果说明遗传多样性水平与物种本身特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低。     

内蒙古地区刺叶柄棘豆(Oxytropis aciphylla Ledeb.)种群遗传多样性分析

用ISSR分子标记对内蒙古地区刺叶柄棘豆(Oxytorpis aciphylla Ledeb.) 5个地理种群进行了种群遗传多样性分析。结果表明：刺叶柄棘豆种群具有较高的遗传多样性,11个ISSR引物扩增出215条带,总的多态位点百分率为98.14%,Shannon多样性指数I=0.2108,Nei基因多样性指数 H=0.341 6,种内总基因多样性(H_t) 为0.2108,种群内基因多样性(H_s)为0.160 4, 大部分遗传变异(76.11%)的遗传变异存在于种群内, 23.89%的遗传变异存在于种群间。遗传分化系数(Gst)为0.238 9,基因流(Nm)为1.592 9。5个居群间已有遗传分化趋势,遗传漂变不会引起遗传分化。UPGMA遗传距离聚类结果表明, 5个地理种群中,植被类型为荒漠草原的4个种群之间遗传距离较近,与1个荒漠种群距离较远。     

入侵植物薇甘菊种群的遗传分化

利用简单重复序列区间(Inter simple sequence repeat, ISSR)分子标记技术分析了入侵植物薇甘菊(Mikania micrantha)8个种群的遗传多样性及遗传分化。12个引物共扩增出171个位点,其中多态位点有103个,多态位点百分率(P%)为60.23%,Shannon信息指数(I)为0.281 8,Nei指数(h)为0.184 9,薇甘菊在物种水平具有较高的遗传多样性。AMOVA显示薇甘菊具有较高的遗传分化,36.49%的变异发生在种群间,63.51%的变异发生于种群内,基因分化系数(G_ST)为0.352 4。种群间的基因流较高,为0.918 7。薇甘菊8个种群之间的遗传相似性很高,平均为0.915 5;遗传距离很小,平均为0.088 4。采用UPGMA法对8个种群进行聚类,可以将8个种群分为两大类群,即内伶仃岛为一个类群,而深圳与东莞内陆种群组成另一类群。薇甘菊现有遗传结构的形成与其生活史特性及入侵生态学特性有关。     

Genetic diversity of Shiraia bambusicola from East China assessed using ISSR markers

To understand the genetic differentiation of the medicinal fungus Shiraia bambusicola, the genetic diversity of 107 individuals from eight populations collected from Jiangsu, Anhui and Zhejiang provinces were studied using inter-simple sequence repeat (ISSR) analysis. The results revealed that the 11 employed primers produced a total of 241 ISSR loci, of which 240 loci (PPB = 99.6%) were polymorphic. Both Nei's gene diversity indexes and Shannon's gene diversity indexes showed that the genetic differentiation of S. bambusicola primarily occurred within the populations. AMOVA revealed that the variation among populations was 40.0%, and the variation within populations was 60.0%. The results of an unweighted pair group method arithmetic average (UPGMA) analysis and a principal coordinate analysis (PCA) revealed that the populations with minimal geographic separations frequently exhibited regional characteristics. These findings revealed that the relationship between sibship and geographical distribution was intensive.     

Genetic diversity and gene flow estimation in Prosopis cineraria (L.) Druce: A key stone tree species of Indian Thar Desert

Selected amplicon data obtained through our earlier study using ISSR and DAMD markers were utilized for determination of diversity within and among the populations of Prosopis cineraria (L.) accessions collected from different districts of Rajasthan (India). A total of 83 bands were generated from eight ISSR and five DAMD primers of which 79 were found to be polymorphic (95.18%). Nei’s gene diversity (h) ranged between 0.185 and 0.301 with overall diversity of 0.316 while Shannon’s information index (I) values recorded between 0.253 and 0.438 with an average value of 0.243. The gene flow value (1.713) and the diversity among populations (0.226) demonstrated higher genetic variation within the population. It is concluded that P. cineraria is accompanied by high genetic diversity within the population and elevated gene flow showing indications of adaptation to callous and fragile dry conditions of arid environment.     

华南及邻近地区大刺鳅遗传多样性的ISSR分析

为了解我国大刺鳅野生资源状况, 研究采用ISSR技术分析了福建、广东、广西、云南和海南11个地理群体262尾大刺鳅的遗传多样性。10个引物共扩增出112个位点, 多态位点95个, 多态位点比例为84.82%。大刺鳅总群体Nei基因多样性h=0.2126、Shannon 信息指数I=0.3358, 表明大刺鳅总群体遗传多样性较丰富, 其中各群体遗传多样性依次为恩平 红河 屯昌 英德 河池 乐昌 增城 龙岩 五华 仁化 百色。总群体遗传分化系数Gst=0.4620, 显示46.2%的变异来自群体间。总群体基因流Nm=0.5823, 表明大刺鳅总群体间缺乏有效的基因交流, 遗传漂变是大刺鳅群体遗传分化的主要因素。聚类分析表明, 东江群体和韩江群体聚为一支, 西江群体和北江群体聚为一支, 大陆群体聚为一大支, 然后和海南屯昌群体聚类。     

木荷种群在演替系列群落中的遗传多样性

利用ISSR分子标记对木荷种群在3个演替系列群落中的遗传多样性进行了研究。12个随机引物共检测到203个可重复的位点,其中多态位点183个,总多态位点百分率（P）为90.15%,平均多态位点百分率为82.27%。Shannon信息指数（I）估算的总遗传多样性为0.524 4,平均为0.477 8。Nei指数（h）计算的总基因多样性为0.358 7,平均为0.326 5。3个种群的P、I、h大小顺序均为针叶林>针阔混交林>常绿阔叶林。AMOVA分子变异显示91.56%变异来源于种群内,8.44%变异来源于种群间。种群间的遗传分化系数（G_ST）为0.089 7,基因流（N_m）为5.073 1。种群间的遗传相似度平均为0.928 4,遗传距离平均为0.074 4,针叶林种群与针阔混交林种群遗传相似度最高。     

Genetic diversity analysis of Hepatacodium miconioides at different altitude in Tiantal Mountain,Zhejiang Province,China and its relationship with environmental factors

The genetic diversity within and among populations of Hepatacodium miconioides collected at three different altitudes in Tiantai Mountain,Zhejiang Province and its relationships to environmental factors were analyzed by random amplified polymorphic DNA(RAPD)technique.Amplification using 12 random primers of 60 plants and 122 repetitive loci were produced.The percentage of polymorphic loci of three populations ranged from 18.85% to 23.77% with an average of 21.86%,indicating the relatively low genetic diversity of H.miconioides.The average Shannon index of phenotypic diversity(0.1329)and Nei index(0.0925)within populations were relatively low.A distinct genetic differentiation existed among populations Of H miconioides in spite of the relatively small geographical distribufion.The average genetic diversity within populations of H.miconioides accounted for 33.58% of the total genetic diversity while the genetic diversity among populations accounted for 66.42% as estimated by the Shannon index of phenotypic diversity,The genetic differentiation among populations of H.miconioides was 0.6546,as estimated by Nei index.The gene flow estimated from Gsr was only 0.2656 and it indicated that gene flow among populations of H.miconioides was relatively low.The mean value of the genetic identity among populations of H.miconioides was 0.7126 and the average of genetic distance of H.miconioides was 0.3412.The genetic identity between populations at the elevation of 990m and at the elevation of 780 m was the highest.The genetic identity between population at the elevation 500 m and other two populations was relatively low.The correlation analysis showed that the genetic diversity within populations was significantly related with the soil total nitrogen.     

A study of conservation genetics in Cupressus chengiana, an endangered endemic of China, using ISSR markers

ISSR markers were used to analyze the genetic diversity and genetic structure of eight natural populations of Cupressus chengiana in China. ISSR analysis using 10 primers was carried out on 92 different samples. At the species level, 136 polymorphic loci were detected. The percentage of polymorphic bands (PPB) was 99%. Genetic diversity (He) was 0.3120, effective number of alleles (Ae) was 1.5236, and Shannon's information index (I) was 0.4740. At the population level, PPB = 48%, Ae = 1.2774, He = 0.1631, and I = 0.2452. Genetic differentiation (Gst) detected by Nei's genetic diversity analysis suggested 48% occurred among populations. The partitioning of molecular variance by AMOVA analysis indicated significant genetic differentiation within populations (54%) and among populations (46%; P < 0.0003). The average number of individuals exchanged between populations per generation (Nm) was 0.5436. Samples from the same population clustered in the same population-specific cluster, and two groups of Sichuan and Gansu populations were distinguishable. A significantly positive correlation between genetic and geographic distance was detected (r = 0.6701). Human impacts were considered one of the main factors to cause the rarity of C. chengiana, and conservation strategies are suggested based on the genetic characters and field investigation, e.g., protection of wild populations, reestablishment of germplasm bank, and reintroduction of more genetic diversity.