用随机扩增多态性DNA(RAPD)分析斯里兰卡龙竹和奥克兰脱竹种群的遗传多样性及其关系

在选择可用于种植的竹种时,必须要了解种群中的异质性。利用随机扩增多态性DNA(RAPD)研究斯里兰卡龙竹和奥克兰脱竹种群间的遗传多样性。龙竹是1856年作为单株植物引种到斯里兰卡,遗传多样性相对较低,仅有0.092±0.027。虽然利用播种的方式进行种种植较为罕见,但用这种方式种植的龙竹很高的遗传距离,因而有较高压的遗传多样性。该种主要是无性繁殖,单株群体没有多态性的无性繁殖系。奥克兰脱竹是当地每年开花的竹种,其平均遗传距离较高,是0.446±0.210。     

珙桐天然种群遗传多样性的ISSR标记分析

利用ISSR分子标记分析来自11个天然珙桐种群的遗传多样性。从100条引物中筛选出5条引物能扩增出稳定、清晰且具多态性的条带,共扩增出77个条带。其中74个为多态,多态条带百分率(PPB)为96.10%;各种群PPB值为37.66%～63.64%,平均为54.07%。种内Shannon多样性指数(HSP)为0.4849,种群内Shannon多样性指数(HPOP)为0.1886～0.3274,平均为0.2774。这表明珙桐在物种和种群水平上均维持较高的遗传多样性。分子方差分析显示,种群间与种群内遗传变异分别占总遗传变异的46.22%,53.78%,种群间呈高度遗传分化。种群间遗传距离与对应的地理距离呈显著正相关(r=0.546,P<0.01)。UPGMA法聚类分析将11个珙桐种群分为3组。研究结果为珙桐遗传资源保护策略制定提供有价值的种群遗传学信息。     

绿竹等30个竹种ISSR遗传多样性分析及指纹图谱构建

为了研究30个竹种的遗传多样性和亲缘关系,建立其DNA指纹图谱,为竹亚科部分植物的分类及种质鉴定提供参考,利用优化的ISSR-PCR体系,筛选出18个多态性较好的的引物,对30个竹种的DNA进行PCR扩增。结果显示,18条引物扩增出的多态性位点占100%,平均每个引物扩增出14个位点,且DNA片段长度在200—3 000bp之间;30个竹种间的平均遗传距离为0.441 4,平均遗传相似度为0.644 44;在遗传距离0.51处进行ISSR聚类分析,可以将竹种分成6组,这与传统的分类大多比较吻合。说明30个竹种具有相当丰富的遗传多样性和基因库,各竹种间有一定的遗传差异和亲缘关系;ISSR技术分析竹种间亲缘关系和遗传多样性具有较高的精确性,可作为竹种分类的参考技术。     

基于ISSR标记的钩栗遗传多样性分析

利用ISSR分子标记对钩栗Castanopsis tibetana Hance 9个野生居群进行了遗传多样性分析。利用100条引物分别对111份钩栗基因组DNA进行PCR扩增,共扩增出158条清晰条带,其中多态性条带141条,多态性条带百分率(P)平均为89.01%。钩栗9个居群的Nei’s基因多样性指数、Shannon’s信息指数分别为0.332 3、0.492 0;总种群基因多样度(Ht)、各种群基因多样度分别为(Hs)0.410 8、0.334 6,表明钩栗的遗传多样性较高。9居群的基因分化系数Gst为0.185 5,占总遗传多样性的81.45%。分子方差分析表明,80.21%的遗传差异存在于群体内,19.79%的遗传差异存在于群体间。基因流为2.196 0。用NTSYS软件计算出9居群111材料的DICE相似系数在0.811 6~0.921 8之间,遗传亲缘关系较近。各居群间的遗传距离差异较大;其中,邵武与建瓯居群的遗传距离最近,仅为0.081 5;浏阳和周宁居群的遗传距离最远,为0.198 4。聚类分析结果表明,采自福建的邵武、建瓯、建阳、屏南和周宁居群聚在一起;恩施和桑植居群聚在一起;永顺和浏阳居群聚为一起,种群遗传变异分布模式基本上与其地理生态格局一致。研究结果表明:供试的钩栗具有较高的遗传多样性,存在着较为频繁的基因交流;基于ISSR标记分析能较准确地揭示出钩栗种间的遗传多样性。     

云南特有濒危植物馨香木兰的遗传多样性研究

为研究云南特有濒危植物馨香木兰的遗传多样性水平,采用随机扩增多态DNA技术（ RAPD）,对云南省馨香木兰4个居群,即文山州广南县、西畴县新街、西畴县亚坪等3个天然居群及昆明树木园1个人工居群,进行遗传多样性分析。随机筛选出重复性好的8条随机引物,对70个样品的基因组DNA进行扩增,建立RAPD分析图谱。扩增结果为,共检测出84个多态位点。遗传多态位点百分率为61.90％（文山广南）、76.19％（西畴新街）、71.43％（昆明树木园）和66.67％（西畴亚坪）。遗传多样性Shannon指数为0.5686,其中30.55％来自种群间,69.45％来自种群内; Nei’s指数为0.3903,种群间的遗传分化系数（GST）为0.2216。文山广南和西畴亚坪之间的遗传距离最大,昆明树木园与西畴亚坪之间的遗传距离最小。结果显示,与木兰科等其他濒危植物相比较,馨香木兰遗传多样性相对较高。根据各种群遗传背景研究结果,提出了该物种迁地保护种群遗传资源管理建议。     

黄土高原不同气象因子对中国沙棘亚居群遗传多样性的影响

利用ISSR分子标记技术,分析了甘肃省东部地区8个中国沙棘居群的遗传多样性和遗传分化,以及气象因子对它们的影响.共调查了8个居群的240个个体.利用11条引物扩增出165条带.扩增范围集中在300～1500 bp,其中157个位点表现为多态,占95.76%,分子变异分析(AMOVA)显示,居群内遗传分化程度高,76.15%的变异存在于居群内.基因分化系数(Gst)为0.2418,居群间基因流为1.5675.说明保护沙棘种群的遗传资源应优先考虑种群内个体.经Mamel检验发现.8个居群的地理距离和遗传距离显著相关(r=0.65,P=0.002).对气象因子与遗传多样性所作的回归分析显示,中国沙棘开花期间的风速与其遗传多样性呈显著正相关.说明开花期风速和地理距离是沙棘种群遗传多样性的重要影响因子.     

四川不同地区慈竹和硬头黄遗传多样性的RAPD分析

试验采用RAPD-PCR手段分别对四川省不同地区的慈竹(Neosinocalamus affin)和硬头黄(Bambusa rigida)进行了遗传多样性研究.结果表明,利用改良的SDS法成功地从硅胶干燥的幼嫩竹叶中提取了质量较高的DNA.用随机引物A9、B17、C2、C5、C11、D3、Q12对8个待测样品扩增结果的多态性分析表明:共获得50个扩增位点(DNA扩增片段),42个位点具有多态性,高达84%,平均每条引物扩增出7.1条带,包括6条多态性带.通过Popgen32软件对8个竹样进行遗传距离分析,同一竹种不同地区之间遗传距离约为0.127 8～0.653 9,这表明不同地区之间存在丰富的遗传多样性;慈竹和硬头黄之间遗传距离较远,为0.446 3～0.916 3.     

地涌金莲野生与栽培种群遗传多样性RAPD分析

利用RAPD分子标记技术对采自滇川两省的12个地涌金莲野生和栽培种群进行遗传多样性分析.选择10条随机引物在12个种群中共扩增出88条带,其中多态性带85条,整个种的多态性位点百分比PPB为96.59%,遗传多样性指数H为0.289 0以及Shannon信息指数I为0.441 6.种群间的遗传分化系数Gst为0.568 2,即43.18%的遗传变异来自于种群内,56.82%的遗传变异来自于种群间,种群间的遗传分化水平略高于种群内.种群间遗传一致度变化范围为在0.66～0.95之间.聚类结果显示:野生种群之间遗传距离较近,与地理分布基本相一致;栽培种群遗传距离较远,与地理分布不一致.     

岛屿地理隔离对山茶种群遗传结构的影响

利用简单重复序列间扩增(ISSR)分子标记对分布于我国浙江和山东的8个山茶种群共240个个体进行了遗传结构分析。结果表明:筛选出的20条引物扩增得到210条清晰条带,其中184条为多态性条带,多态位点百分率(PPB)为87.62%。经POPGENE1.32软件分析,山茶种群平均多态位点百分率(PPB)为68.99%,Nei’s基因多样性指数(HE)为0.256 9,Shannon信息指数(H)为0.377 9,种群遗传多样性较高。基因分化系数Gst=0.182 9,表明遗传变异主要存在于种群内个体间。地理距离与遗传距离具有显著相关性(r=0.856 7,P<0.05),岛屿地理隔离对山茶种群的遗传分化具有重要影响。UPGMA聚类表明:浙江5个山茶种群间的遗传相似度较高,山东青岛的植物园种群和五四广场种群可能移植自长门岩岛。我国野生山茶资源受人为破坏严重,建议在加强现有岛屿自然种群就地保护力度的同时,建立山茶种质资源库,促进基因交流。     

乐昌含笑种群遗传多样性的研究

基于随机扩增多态DNA(RAPD)方法分析了珍稀木兰科植物乐昌含笑6个种群的遗传多样性及分化程度。19条随机引物扩增出111个可分析位点,多态位点百分比为81．98％,经POPGENE分析发现,乐昌含笑的Nei’s基因多样度(Hc)为0．3255,Shannon表型指数(I)为0．4751,与其它植物比较具有较高水平的遗传多样性。6个乐昌含笑群体间基因分化系数(Gst)值为0．2226。群体内的遗传变异大于群体问的遗传变异。按照遗传距离将乐昌含笑6个群体划分为两类：第1类有广西三江、湖南宜章、湖南双牌和湖南资兴的群体,第2类包括江西官山和湖南醴陵的群体;研究表明：第2类种群的遗传多样性指数高于第1类种群的遗传多样性指数。     

Analysis of genetic diversity for wild and captive green peafowl populations by random amplified polymorphic DNA technique

The genetic diversity of the populations for 14 wild green peafowls (Pavo muticus) and 18 captive green peafowls was investigated by using the technology of random amplified polymorphic DNA (RAPD). Totally 161 and 166 amplified bands were obtained by using 23 arbitrary primers to amplify the genomic DNA of wild and captive green peafowls respectively. The results showed that the average relative genetic distance of the wild and captive green peafowls populations was 0.0555 and 0.1355, respectively, and difference of the average relative genetic distances between the two populations was 0.1635. The Shannon diversity index for the wild and captive green peafowl populations was 0.4348 and 1.0163, respectively, which means that there exists significant difference in genetic diversity between the two populations, and the genetic diversity of wild green peafowl was low. The two populations originated from two different families according to analysis by the UPGMA method. This research can provide the theoretical basis for supervising genealogies management of peafowl populations.     

绿孔雀在中国的现状及保护策略

绿孔雀曾分布于中国南方多个省份,由于人类活动等因素的影响,绿孔雀分布区不断退缩,野生种群数量持续下降。目前绿孔雀在中国仅见于云南省,呈斑块化分布,数量约500只,已变得罕见。根据相关资料,从绿孔雀生物学特性、野生种群数量、分布区现状、种群结构和种群隔离几方面分析中国绿孔雀面临的灭绝风险,在就地保护的前提下,重点探讨易地保护策略。提出人工繁育,建立纯绿孔雀人工种群用于开展野外放归,逐步扩大绿孔雀分布区,增加野生种群数量,降低灭绝风险等保护策略。     

撑篙竹遗传变异的RAPD分析

采用随机扩增多态DNA(RAPD)方法对6个群体30丛撑篙竹个体进行了遗传变异的研究。28个随机引物共检测到173个位点,其中85个是多态位点,平均每个引物提供6．18个RAPD信息量,扩增出的DNA片段大小一般在200～2000bp范围之间;用POPGENE1．31版软件进行遗传多样性分析：平均Nei’s基因多样性为0．2114,Shannon’s信息指数为0．3277,基因分化系数0．1853,表明群体间有一定的分化;各群体平均遗传距离0．0350,表明群体亲缘关系较近;试用UPGMA方法对不同产地的撑篙竹群体作聚类分析,初步可将6个群体聚为3类。     

Genetic diversity and population structure analysis of Pistacia species revealed by phenylalanine ammonia-lyase gene markers and implications for conservation

Information on population genetic structure and crop genetic diversity is important for genetically improving crop species and conserving threatened species.The PAL gene sequence is part of a multigene family that can be utilized to design DNA marker systems for genetic diversity and population structure investigation.In the current study, genetic diversity and population structure of100 accessions of wild Pistacia species were investigated with 78 PAL markers.A protocol for using PAL sequences as DNA markers was developed.A total of 313 PAL loci were recognized, showing 100% polymorphism for PAL markers.The PAL markers produced relatively more observed and effective alleles in Pistacia falcata and Pistacia atlantica, with a higher Shannon's information index and expected heterozygosity in P.atlantica, Pistacia vera and Pistacia mutica.Pairwise assessment of Nei's genetic distance and genetic identity between populations revealed a close association between geographically isolated populations of Pistacia khinjuk and Pistacia chinensis.The accessions of wild Pistacia species had more genetic relationship among studied groups of species.Analysis of molecular variance indicated 19% amongpopulation variation and 81% within-population variation for the PAL gene based DNA marker.Population structure analysis based on PAL revealed four groups with high genetic admixture among populations.The results establish PAL markers as a functional DNA marker system and provide important genetic information about accessions from wild populations of Pistacia species.     

Genetic diversity analysis of Sinojackia microcarpa,a rare tree species endemic in China,based on simple sequence repeat markers

Although most Sinojackia species are endangered, they contribute greatly to the biodiversity of local ecosystems. Sinojackia microcarpa, an endangered species, is distributed only in three provinces of eastern China. Determining the genetic diversity of S. microcarpa provides key information for germplasm evaluation and species conservation. Here we used simple sequence repeat (SSR) markers to investigate the genetic diversity of eight natural populations of S. microcarpa. Leaf samples were collected from 144 individuals in 8 wild populations. The 156 bands were generated from 14 pairs of informative SSR primers, with an average percentage of polymorphic bands of 45.67%. The average values of Nei’s genetic diversity (He) and Shannon’s diversity index (I) were 0.1007 and 0.1658, respectively. The total genetic variation of S. microcarpa existed mainly within the eight populations, rather than among populations, and reached 86.41%. A cluster analysis showed that the eight wild populations of S. microcarpa could be classified into four groups, at a threshold of 4.0, based on an analysis of the SSR genotypes. Furthermore, there was a significant association between the phylogenetic relationships and the geographic locations of the S. microcarpa populations. In particular, populations from Fuyang, Jiande, and Lin’an in Zhejiang Province had close phylogenetic relationships and geographic distances. In addition, these three populations had the highest genetic diversity and the most individuals, suggesting that these three locations may be the S.microcarpa distribution center. This study serves as a model for studying the genetic diversity of endangered plant species.     

成都麻羊种群微卫星标记遗传多样性评估

作为优良的地方山羊品种,成都麻羊对我国山羊品种的改良和新品种培育起了重要作用。畜禽遗传资源本质上是基因资源,保护畜禽遗传资源就是保护基因的多样性。为了了解成都麻羊的遗传情况,通过微卫星标记技术,选用8对中高度多态微卫星标记进行成都麻羊圈养种群的遗传多样性评估,共检测到60个等位基因,平均等位基因和平均有效等位基因数分别为7.5、2.36,平均观察杂合度、平均期望杂合度和平均多态信息含量分别为0.577、0.634和0.580。研究结果表明成都麻羊种群遗传多样性较高,具有较高的保种潜力。     

湖北三潭风景区野生省沽油遗传多样性的ISSR分析

采用ISSR分子标记技术对湖北三潭风景区野生省沽油群体32个样本进行遗传多样性分析,从18条ISSR引物中筛选出10条引物用于PCR扩增,共扩增出147条DNA条带,其中多态性条带134条,占91.16%,平均每个引物扩增条带的数目为14.7条。利用POPGENE1.32和NTSYS-pc软件分析得出该野生省沽油样本的平均有效等位基因数为1.4319,平均Nei′s基因多样性指数为0.2653,平均Shannon信息指数为0.4118;样本间的遗传相似系数在0.47～0.87之间;UPGMA聚类结果,在遗传相似性系数0.6558处可把供试的32个样本分为3大类,表明该野生省沽油群体的遗传多样性比较丰富。     

Genetic diversity of natural Hepatacodium miconioides populations in Zhejiang Province

Hepatacodium miconioides is the Class II protected plant species in China. This paper studies the genetic diversity and differentiation of its nine natural populations in Zhejiang Province by using random amplified polymorphic DNA (RAPD) technique. Twelve random primers were selected in the amplification, and 164 repetitive loci were produced. The percentage of polymorphic loci in each H. miconioides population ranged from 14.60% to 27.44%, with an average of 20.73%. Among the test populations, Kuochangshan had the highest percentage of polymorphic loci, Simingshan took the second place, and Guanyinping had the lowest percentage. As estimated by Shannon index, the genetic diversity within H. miconioides populations accounted for 27.28% of the total genetic diversity, while that among H. miconioides populations accounted for 72.72%. The genetic differentiation among H. miconioides populations as estimated by Nei index was 0.715,7. This figure was generally consistent with that estimated by Shannon index, i.e., the genetic differentiation among populations was relatively high, but that within populations was relatively low. The gene flow among H. miconioides populations was relatively low (0.198,7), and the genetic similarity ranged from 0.655,7 to 0.811,9, with an average of 0.730,6. The highest genetic distance among populations was 0.422,9, while the lowest was 0.208,3. All the results showed that there was a distinct genetic differentiation among H. miconioides populations. The genetic distance matrix of nine test populations was calculated using this method, and the clustering analysis was made using the unweighted pair group method with arithmetic mean (UPGMA). The cluster analysis suggested that the nine populations of H. miconioides in Zhejiang Province could be divided into two groups, the eastern Zhejiang group and the western Zhejiang group. __________ Translated from Chinese Journal of Applied Ecology, 2005, 16(5): 795–800 [译自: 应用生态学报, 2005, 16(5): 795–800]     

湘鄂西地区珙桐天然群体遗传结构的研究

以珙桐叶为材料,对湘西和鄂西两天然珙桐群体的60个样品进行RAPD分析.结果表明,9个随机多态引物,获得24个多态标记位点,平均每个引物产生2.67个多态位点;湘西和鄂西两群体之间的遗传多样性差异不明显,Shannon指数分别为0.427 4和0.448 1;种内平均遗传多样性为0.326 9,群体内平均遗传多样性为0.297 6,群体间的遗传多样性为0.029 3,基因分化系数为0.089 6,基因流为5.079 2.两个天然珙桐群体的遗传相似性非常高,达到0.923 3,遗传距离为0.079 8;湘西群体的多态百分位率略高于鄂西.这表明,湘西和鄂西两群体间存在着非常频繁的基因流.     

基于表型和ISSR标记滇产黄杨叶栒子遗传多样性分析

[目的]从表型和分子2个角度揭示黄杨叶栒子(Cotoneaster buxifolius Lindl.)野生种群的遗传多样性。[方法]利用33个表型性状和ISSR分子标记对滇产野生黄杨叶栒子进行了遗传多样性分析。用PopGen 32软件计算多样性指数,用SPSS 16.0软件构建系统发育树系图。[结果]黄杨叶栒子种内表型性状在居群间存在着丰富的变异,居群间表型性状叶片大小变异系数达25.99%,萼裂片相关3个性状变异达40.66%,差异较大;24条ISSR引物扩增出228条带,其中,153条具有多态性,平均多态率为81.56%,Nei’s基因多样性指数和Shannon多样性指数分别为0.352 0和0.505 9,具有较高的遗传多样性;采用UPGMA法构建遗传关系聚类图,形态和分子标记的2种聚类结果基本一致:分布在同一地区的各个居群多数能聚在一起,地理位置的隔离促进了黄杨叶栒子居群间的遗传分化;个别居群未能与同一产区的其他居群聚在一起,而聚进了其他产区的居群之中,其原因可能是同一产区内各居群的小生境不完全相同,在不同产区内也有相似的小生境,导致黄杨叶栒子居群间发生了趋同进化。[结论]黄杨叶栒子遗传多样性丰富,居群间的亲缘关系及分布与地理位置、海拔位置有密切关系。