沿海岛屿与内陆藓类植物遗传多样性和分化的比较研究——以真藓(Bryum argenteum Hedw.)为例

基于11个内陆居群和32个岛屿居群的252份标本,采用ISSR分子标记技术对真藓（Bryum argenteum Hedw.）的遗传多样性进行了研究。结果显示：岛屿与内陆居群间的遗传多样性差异显著,岛屿居群间的分化程度（G_st=0.453）大于内陆居群（G_st=0.387）,岛屿居群的遗传分化与地理来源间存在极显著相关性（r=0.478,n=175,P<0.001）。地理隔离效应是导致岛屿居群间遗传分化的重要因素。岛内居群间的遗传分化水平较低,仅有29.4%~29.7%的遗传多样性存在于居群间。聚类分析表明,43个居群可划分为10大类群,真藓遗传关系受地理因素和生境异质性的影响,水域隔离影响了真藓繁殖体在岛屿间的传播。     

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     

利用rp132-trnL分析阿尔金山自然保护区真藓居群遗传多样性

本研究对分布在中国新疆维吾尔自治区阿尔金山自然保护区的6个真藓(Bryum argenteum)居群的遗传结构及遗传多样性进行比较。通过对32条叶绿体DNA的rpl32-trnL序列的核苷酸序列变异的分析,发现了14种单倍型,存在201个可变位点;分子变异分析显示有51.02%的遗传变异发生在居群间水平,居群内部的遗传变异为48.98%,真藓居群间的遗传分化程度较高略大于居群内的遗传分化。居群遗传变异的分化系数为0.510 2,基因流值为0.48,显示各居群间的基因流低。单倍型多态性水平为(0.780 2±0.076 0),核苷酸多态性水平为(0.058 59±0.020 09),表明真藓居群遗传多样性丰富。对所有变异位点进行的Taijma's检验的结果是Taijma's D值为-1.567 05 (p0.10),显示所有变异符合中性进化假说。     

濒危特有种掌叶木的微卫星遗传多样性研究

掌叶木居群具有较丰富的遗传多样性,该研究利用9对微卫星(SSR)分子标记揭示了掌叶木(Handeliodendron bodinieri)的遗传多样性。结果表明:观测等位基因数(Na)平均为3.903,有效等位基因数(Ne)平均为2.545,期望杂合度(He)平均为0.521,Shannon’s多态性信息指数(I)为0.962,PIC平均值为0.465。掌叶木的自然分布居群有相对较高的遗传多样性,但由于人为破坏等因素导致该群体濒危,而濒危并不是因为遗传多样性降低而造成的。居群间的遗传分化为掌叶木8个居群间的遗传一致度为(GI=0.849~0.970),遗传距离为(GD=0.032~0.164)。基于Nei’s遗传距离用UPGMA法对掌叶木居群进行聚类,Nei’s的基因分化系数为(G_(st))为0.027,平均Nei标准遗传分化系数(G'_(st)N)为0.031,平均Herick’s标准遗传分化系数(G'_(st)H)为0.064,基因流(N_m)为3.368。AMOVA分析结果表明:掌叶木居群间变异占3%,居群内变异占97%,居群内的遗传分化大于居群间的分化。利用Mantel检测发现,居群间的遗传距离与地理距离显著正相关(r=0.299,P0.05)。该研究结果为掌叶木生物多样性和资源保护与利用提供了更充分的科学依据。     

rpl32-trnL片段在柳穿鱼不同地理居群遗传多样性分析中的应用

为了探究柳穿鱼(Linaria vulgaris)不同地理居群的遗传多样性,利用叶绿体DNA的rpl32-trnL片段对包含62个个体的4个柳穿鱼地理居群遗传多样性进行了研究。结果显示：柳穿鱼4个居群中共检测到15种单倍型和76个变异位点,总遗传多样性较高(Hd=0.878,π=0.003 88,K=2.994),遗传变异主要存在于居群内(51.49%),隶属于柳穿鱼虫媒异交繁殖策略的遗传特征;不同地区柳穿鱼居群间遗传分化大(0.466 14),居群间基因交流水平较低(0.29);遗传分化程度与地理距离存在中等程度相关性但不显著(R²=0.36,P> 0.05)。中性检验显示除合水居群(HS)在进化过程中经历过瓶颈效应(Fu and Li’s D=-2.450 49,P<0.05)外,其他居群进化过程符合分子进化的中性理论。本研究结果不仅揭示了繁殖策略、地理隔离及生境干扰等因素塑造我国北方柳穿鱼居群遗传多样性和遗传结构特征,而且也为今后柳穿鱼资源保护策略的选择提供了理论依据。     

湖北野生天麻的遗传分化及栽培天麻种质评价

采用7条ISSR引物对天麻(Gastrodiaelata)8个自然居群和6个人工栽培居群共483个样本的居群遗传多样性进行了初步检测,共检测出清晰、重复性好的DNA带77条,其中64条为多态性带,总多态位点百分比PPB=83.12%。遗传多样性分析结果表明:天麻自然居群的遗传多样性参数分别为:多态位点百分比PPB=59.09%,有效等位基因数Ae=1.29,Nei’s遗传多样度H=0.176,Shannon’s多态信息指数I=0.270,明显高于人工栽培居群(PPB=35.71%,Ae=1.16,H=0.100,I=0.155),揭示出栽培居群存在明显的遗传基础狭窄和遗传均质性问题。UPGMA聚类分析表明,自然居群与栽培居群存在明显的分化而分别聚为两大类群。自然居群间基因分化系数GST=0.2558,与AMOVA分析所揭示的居群间遗传变异量占总变异的27.25%的结果相近,说明天麻自然居群间亦存在一定程度的遗传分化;居群间基因流(Nm)为1.4547,相对较弱,可能对自然居群的遗传分化有一定影响。自然居群聚类结果显示出一定程度的地理区域聚类趋势,但Mantel检验表明自然居群间遗传距离与地理距离并不存在显著相关(r=0.1669,P=0.2110),揭示出天麻自然居群的分化现状可能是其生活史特性、地理隔离与人为破坏综合作用的结果。栽培居群的遗传均质化趋势,揭示了引种驯化的瓶颈效应和长期无性繁育所导致的遗传多样性丧失,也反映出栽培天麻种质的遗传基础狭窄。而栽培居群与自然居群间存在着明显的遗传分化,反映天麻栽培居群与自然居群间可能存在基因流的阻断。     

宝兴百合与匍茎百合遗传多样性的ISSR分析

利用5条ISSR引物对宝兴百合(Lilium duchartrei)9个居群和匍茎百合(Lilium lankongense)13个居群的遗传多样性进行了初步检测。结果表明:(1)宝兴百合在物种水平上多态位百分率(PPB)为97.26%,Nei’s基因多样度(H)为0.309 8,Shannon’s多样性信息指数(Hsp)为0.469 4;匍茎百合在物种水平上多态位百分率(PPB)为100%,Nei’s基因多样度(H)为0.3390,Shannon’s多样性信息指数(Hsp)为0.503 0,均略高于宝兴百合。(2)宝兴百合与匍茎百合的遗传多样性在居群水平上相对较低;宝兴百合和匍茎百合居群间遗传分化系数(Gst)分别为0.642 5和0.563 7,表明2个物种居群间的遗传分化大于居群内的遗传分化。(3)经Mantel检测,两种居群间的遗传距离与地理距离均不存在显著的相关性(宝兴百合r=0.263 7,P=0.844 0;匍茎百合r=0.104 2,P=0.695 0);宝兴百合与匍茎百合的遗传多样性及遗传分化现状可能是两者的生活史特性、地理隔离等作用的结果。(4)UPGMA聚类结果显示,宝兴百合与匍茎百合在分子水平上出现了明显分化,支持二者是独立的物种。     

濒危植物毛瓣金花茶遗传多样性的ISSR分析

采用ISSR分子标记对毛瓣金花茶6个自然居群的遗传多样性进行了分析。利用11个引物对150个个体进行了扩增,共扩增出92条条带,其中多态性条带74条。毛瓣金花茶在物种水平和居群水平都表现出相对较高的遗传多样性,在物种水平上,多态位点百分率(PPB)为80.43%,Nei’s基因多样性指数(h)为0.245 1,Shannon多样性指数(I)为0.377 6;在居群水平上,PPB为58.70%~66.30%,h为0.199 7~0.229 3,I为0.300 9~0.343 8。Nei’s遗传多样性分析和AMOVA分析表明,毛瓣金花茶的遗传变异主要存在于居群内,居群间的遗传分化程度较低(Gst=0.126 6,Φst=11.37%),基因流(Nm)为3.448 0。Mantel检测表明,居群间的遗传距离和地理距离之间存在显著的相关关系(r=0.755 1,P0.05)。研究认为,毛瓣金花茶较高的遗传多样性和较低的遗传分化可能与其异交型繁育系统和鸟类传粉有关。     

我国北回归线区域普通野生稻遗传多样性和遗传结构研究

选择分布于水稻染色体组的26对SSR引物,对集中分布于广西来宾市北回归线附近的13个普通野生稻居群的342份材料进行遗传多样性和遗传结构研究。结果表明,该地区的野生稻无论在种内(A=9.154,Ae=4.446,I=1.547,He=0.671)还是居群水平上(P=95%,A=4.219,Ae=2.394,I=0.905,He=0.476)遗传多样性都十分丰富,高于同类研究水平。供试居群的遗传分化系数Gst为0.30,表明30%的遗传变异存在于居群间,大部分遗传变异存在于居群内。居群间的遗传一致度变化范围为0.332~0.903,且居群间地理距离越近,遗传一致度越高,说明北回归线附近的普通野生稻居群符合"隔离-距离"模型。综合上述研究结果和我国野生稻保护现状,建议对居群G13和G06实施优先保护。     

濒危植物三棱栎遗传多样性的RAPD分析

用随机扩增多态DNA (RAPD)标记对 5个三棱栎 (Trigonobalanusdoichangensis)居群共 99个个体进行遗传多样性和居群遗传结构分析。 16个引物共检测到 15 7个位点 ,其中多态位点 83个 ,占 5 2 87%。物种水平Shannon多样性指数I =0 2 4 31,Nei基因多样度h =0 15 95 ,种内总遗传变异量Ht=0 16 0 0 ,居群内遗传变异量Hs =0 0 74 9,居群间变异量大于居群内变异量 ,表明三棱栎的遗传变异主要存在于居群之间。与同科植物相比 ,三棱栎遗传多样性较低 ,遗传分化系数Gst =0 5 32 0 ,说明居群间的遗传变异占 5 3 2 0 % ,居群间已出现强烈的遗传分化。当地人的强烈活动造成的生境破碎化和居群隔离 ,以及三棱栎演化过程中的地史变化对其种群发展的影响等 ,可能是造成其居群间强烈的遗传分化和较低遗传多样性的原因。基于本研究结果 ,提出了三棱栎遗传多样性的保护策略。     

ISSR and SRAP Markers Reveal Genetic Diversity and Population Structure of an Endangered Slipper Orchid,Paphiopedilum micranthum (Orchidaceae)

Paphiopedilum micranthum is an endangered pink slipper orchid mainly distributed in the limestone areas of southwestern China. Wild populations of this species have been seriously threatened by excessive collections, rampant smuggling for export, and habitat destruction. We used 15 ISSR markers and 11 SRAP markers to investigate the genetic diversity and structure of 15 natural populations. A high degree of diversity was observed at the species level (ISSR: PPB=9166%, He=03839; SRAP: PPB=9929%, He=02806). Certain degree of genetic differentiation among populations (ISSR: Gst=02577; SRAP: Gst=02383) was detected maybe caused by low gene flow (ISSR: Nm=07201; SRAP: Nm=07991). Consistent with the results of Principal Coordinate Analysis, the UPGMA dendrogram analysis divided the 15 populations into two main clades. In addition to geographic distance, the difference in elevation was another natural factor contributing to this differentiation. Knowledge about genetic diversity and structure gained from our study will be beneficial for the development of reasonable and efficient strategies to conserve this endangered species.     

山东省长岛县南部四岛藜（Chenopodium album L.）天然种群遗传多样性的ISSR分析

采用ISSR分子标记对分布于山东省长岛县南部的北长山、南长山、大黑山和小黑山4个岛屿上的藜天然种群共81个个体进行了遗传多样性及遗传结构的研究。13个引物共检测到157个可重复的位点。遗传多样性研究结果表明：种内的多态位点比率为66.24％,具有较高的遗传多样性（Shannon（I）指数在物种水平上为0.332 0）;种群间有一定的遗传分化,根据G_st值,种群间的遗传多样性占总群体的9.27％。遗传距离分析表明,XHS种群和NCS种群的遗传一致度最高,与地理距离无相关性。     

珍稀濒危植物硬叶兜兰的遗传多样性及遗传结构研究

由于人为采集、走私贩卖以及生境的破坏,分布于中国西南石灰岩地区的野生硬叶兜兰居群受到严重的干扰与威胁。为有效地保护这种珍稀野生植物,本研究采用ISSR和SRAP两种分子标记对15个硬叶兜兰野生居群进行遗传多样性及遗传结构的研究。结果表明,硬叶兜兰在物种水平上具有较高的遗传多样性（ISSR：PPB=91．66％,He=0．3839;SRAP：PPB=99．29％,Hc=0．2806）。硬叶兜兰居群间存在一定程度的遗传分化（ISSR：Gs1： 0．2577;SRAP：Gst=0．2383）,可能由于较低的基因流（ISSR：Nm=0．7201;SRAP：Nm=0．7991）所致。UPGMA聚类分析以及主成分分析均把15个居群分成2个主要分支。居群间的地理距离和海拔差距是引起居群遗传分化的自然因素。     

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.     

竹叶眼子菜居群遗传多样性和克隆结构

采用ISSR技术对长江中游南岸豹澥湖和大冶湖不同生境中的竹叶眼子菜(Potamogeton malaianus)居群的遗传多样性及克隆结构进行了研究.结果表明,在两居群的106株个体中,利用6条ISSR引物共得到40条符合3/N标准并无连锁不平衡的清晰位点,竹叶眼子菜具有较高的遗传多样性,其多态位点百分率为75.0％,Shannon多样性指数为0.3736, 两居群的遗传分化很小.竹叶眼子菜的克隆多样性很高(D=0.9917),两居群间的克隆分化很大,不具有共有的基因型.竹叶眼子菜的基株分布为游击型构型,位于湖心的大冶居群克隆距离(3.0～31.5 m)明显地大于湖岸豹澥居群(2.4～6.7 m).     

Population Genetic Diversity of Bombax malabaricum(Bombacaceae) in China

In order to discuss the level of genetic diversity of Bombax malabaricum, we surveyed the genetic diversity of 4 populations (YJ, YM, QJ, DJ) from dry-hot valleys of Yunnan province, 2 populations (GX, HN) from dry-hot regionsand 1 population (BN) from wet-hot region of Yunnan using ISSR molecular markers . Based on 10 selected primers, 142 clear and reproducible DNA fragments were generated. The percentage of polymorphic loci PPB was 90.14%, Nei′s ( 1973) gene diversity H was 0. 2530 and Shannon′s Information index I was 0. 3864 . The coefficient of genetic differentiation( G_ST ) was 0. 1870 and the Фst was 0. 177 estimated by AMOVA. The results showed high level of genetic diversity within population and low level of genetic differentiation among populations. We inferred that the high level of genetic diversity and effective gene flow of B. malabaricum may play an important role in its better adaptability. Considering the introduction of B. malabaricum in dry-hot regions, we suggest to sample abundantly within populations and involve different populations .     

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。     

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 variation among wild and cultivated populations of the Chinese medicinal plant Coptis chinensis (Ranunculaceae)

To examine if the cultivation process has reduced the genetic variation of modern cultivars of the traditional Chinese medicinal plant, Coptis chinensis, the levels and distribution of genetic variation was investigated using ISSR markers. A total of 214 C. chinensis individuals from seven wild and three cultivated populations were included in the study. Seven ISSR primers were used and a total of 91 DNA fragments were scored. The levels of genetic diversity in cultivated populations were similar as those in wild populations (mean PPL = 65.2% versus PPL = 52.4%, mean H = 0.159 versus H = 0.153 and mean I = 0.255 versus I = 0.237), suggesting that cultivation did not seriously influence genetic variation of present-day cultivated populations. Neighbour-joining cluster analysis showed that wild populations and cultivated populations were not separated into two groups. The coefficient of genetic differentiation between a cultivar and its wild progenitor was 0.066 (G(st)), which was in good accordance with the result by amova analysis (10.9% of total genetic variation resided on the two groups), indicating that cultivated populations were not genetically differentiated from wild progenitors. For the seven wild populations, a significant genetic differentiation among populations was found using amova analysis (45.9% of total genetic variation resided among populations). A number of causes, including genetic drift and inbreeding in the small and isolated wild populations, the relative limited gene flow between wild populations (N(m) = 0.590), and high gene flow between cultivars and their wild progenitors (N(m) = 7.116), might have led to the observed genetic profiles of C. chinensis.