不同倍性桤木属植物遗传差异的AFLP分析

采用AFLP技术对桤木属4种不同倍性的116份材料指纹图谱进行了分析,5对引物组合共得到301个位点,282个为多态位点,多态位点百分率为93.7%。4个种群多态位点百分率为35.2%~72.4%,平均多态位点百分率为54.6%。总的Nei's基因多样性为0.3043,总Shannon多态信息指数为0.4578。种群间及种群内部均具有较丰富的遗传多样性,种群间的遗传差异大于种群内部的遗传差异。NJ聚类分析与主坐标分析结果一致,所有的供试材料明显的分为3个组,地理分布距离较近的种群优先聚在一起。     

蜡梅种质资源遗传多样性的ISSR分析

利用ISSR分子标记技术,对蜡梅种质资源7个野生种群和2个栽培种群的遗传多样性进行了研究。用11条引物,共扩增出124条谱带,其中110条多态带,多态位点占88.70％。用POPEGEN1.31版软件对数据进行分析,结果显示：种群总的Nei’s基因多样性指数为0.272 6,Shannon信息多态性指数为0.411 7,蜡梅总的遗传多样性水平较高。蜡梅不同种群遗传多样性水平差异较大,种群多态位点百分率在52.94％~90.00％之间,Nei’s基因多样性指数为0.143 4~0.378 2,Shannon信息多态性指数为0.232 2~0.546 6。神农架种群（SN）和保康种群（BK）的遗传多样性水平较高。种群间的基因分化系数为0.353 6,种群内的遗传变异大于种群间的遗传变异。用NTSYS2.01版软件对样品进行UPGMA聚类分析,结果9个种群并没有按地理距离进行聚类。种群间的地理距离和遗传距离之间没有显著的相关性(r＝0.437 1,P＝0.921 3)。     

秦岭秀雅杜鹃野生种群遗传多样性和遗传分化的AFLP分析

利用AFLP分子标记技术,对秦岭地区7个秀雅杜鹃野生种群的遗传多样性和遗传分化进行研究.结果表明:M52E41、M62E46和M64E94等3个引物组合共扩增出182条DNA片段,其中151条是多态的,多态位点比率为83.1％.7个秀雅杜鹃种群所检测的多态位点百分率(PPL)、Nei的基因多样性指数(h)和Shannon信息指数(I)的变化趋势一致,其排序为眉县种群＞柞水种群＞镇安种群＞户县种群＞宁强种群＞南郑种群＞周至种群.POPGENE分析表明,秀雅杜鹃在物种水平(PPL=91.22％,I=0.7217,h=0.5095)和种群水平(PPL=77.56％,I=0.6409,h=0.4725)都具有较高的遗传多样性.种群间的遗传分化系数(Gst)为7.26％,说明有92.74％的变异存在于种群内部.AMOVA分析表明,在总的遗传变异中,85.3％的变异发生在种群内,14.7％的变异发生在种群间,说明秀雅杜鹃种群的遗传变异主要存在于种群内部.UPGMA聚类分析表明,种群间的遗传距离与种群间的地理距离没有明显的相关性.最后提出了秦岭地区秀雅杜鹃种质资源的保护策略.     

大猿叶虫四地理种群遗传多样性的RAPD分析

以大猿叶虫Colaphellus bowringi Baly 4个地理种群的基因组DNA为材料, 进行RAPD分析。从80条引物中筛选出11条稳定性好、多态性高的引物进行扩增, 共得到65个扩增位点, 53个多态位点, Nei氏遗传多样性指数为0.1049~0.2061, Shannon多样性指数为0.1641~0.3167。结果表明所分析的大猿叶虫遗传变异很高, 其中江西龙南种群遗传变异最小, 山东泰安种群遗传变异最高。种群间的遗传距离范围为0.0636~0.3200, 其中江西龙南种群和江西修水种群间的遗传距离最小, 哈尔滨种群与江西龙南种群间的遗传距离最大, 种群遗传距离的大小与其相对地理距离的远近吻合。结果提示种群遗传距离的大小与它们生物学上的相似性有关联。     

枫香自然种群遗传多样性的ISSR分析

利用ISSR分子标记技术,对浙江省内枫香自然种群的遗传多样性进行分析。用10个引物对5个枫香种群共100个个体的样品DNA进行扩增,共测得135个位点,其中多态位点为118个,多态位点百分率(P)为87.41%,Shannon指数(I)为0.4646,Nei指数(h)为0.3122,表明枫香总体水平的遗传多样性较高。各种群的多态位点百分率平均为59.11%,Shannon指数平均为0.3660,Nei指数平均为0.2543。P、I、h均显示北山种群最高,天台山种群最低。AMOVA分子差异分析显示:85.49%的变异存在于种群内,14.51%存在于种群间,基因分化系数(Gst)为0.1856,种群间的遗传分化程度较低。种群间的基因流(Nm)为2.1944。5个种群间的平均遗传距离为0.1199。利用UPGMA法对5个种群进行聚类分析,结果分为两大类群:白云山、天台山、北山和安岱后4个种群组成一大类群;大明山种群单独为另一类群。     

东南石栎种群在演替系列群落中的遗传多样性

在浙江省天台山,利用RAPD技术对东南石栎种群演替系列群落针叶林、针阔混交林、常绿阔叶林中的遗传多样性和遗传分化进行了研究。结果表明,12个随机引物在60株个体中共检测到173个可重复位点,其中多态位点152个,总多态位点百分率为87.86%,3个种群平均多态位点百分率为65.32%。采用Shannon信息指数计算的3个种群总的遗传多样性为0.4529,平均为0.3458。采用Nei指数计算的3个种群总的基因多样性为0.3004,平均为0.2320。3个种群的多态位点百分率、Shannon信息指数、Nei指数大小顺序均为针叶林种群>针阔混交林种群>常绿阔叶林种群。AMOVA分子变异显示,72.85%变异来源于种群内,27.15%变异来源于种群间。种群间的遗传分化系数为0.2277,基因流为1.6949。东南石栎这种遗传结构是其本身生物学特性的反映,同时也与群落的微环境密切相关。3个东南石栎种群间的遗传相似度平均为0.8662,遗传距离平均为0.1442。针阔混交林种群与常绿阔叶林种群的遗传相似度最高,常绿阔叶林种群与针叶林种群最低。根据Nei的遗传距离对不同种群进行非加权成组配对法(UPGMA)聚类表明,针阔混交林种群与常绿阔叶林种群先聚合,再与针叶林种群聚在一起。     

山稻蝗不同种群的遗传多样性研究

应用RAPD技术对我国长江以南丘陵山区6个种群山稻蝗Oxya agavisa进行了遗传多样性分析.10条随机引物共扩增出90条清晰稳定的片段,其中多态性位点为87个,总的多态位点百分率96.7%.采用Shannon's遗传多样性值和Nei's基因多样性指数分析种群内的遗传多样性及种群间的遗传分化,结果表明山稻蝗湖南通道种群遗传多样性最高;两种指数估算的种群间遗传分化系数(44.5%,41.3%)均表明:山稻蝗种群内的遗传多样性稍大于种群间的遗传多样性.分别用Between-groups linkage、UPGMA和NJ法构建所有个体间的聚类关系图,聚类结果分析表明:位于同一纬度附近且地理上相距较近的种群优先相聚,聚类关系与生境条件、地理距离间呈现出一定的相关趋势.     

重金属污染芒萁居群遗传分化的RAPD分析

利用随机扩增多态DNA标记技术(random amplified polymorphic DNA,RAPD)对自然生长的芒萁洁净居群和铅锌矿尾矿居群的遗传多样性和遗传分化进行分析。结果表明在170个检测位点中发现有102个位点呈多态性,在尾矿居群中有2个RAPD标记位点发生了丢失,推测可能为重金属的敏感基因片段;尾矿居群的多态位点百分率(52.94%)、Shannon信息指数(0.3059)和Nei基因多样性(0.2084)均略低于洁净居群(分别为53.53%、0.3196和0.2198),总的遗传多样性水平较高(分别为70.59%、0.3723和0.2472)。尾矿居群与洁净居群之间发生了一定的遗传分化(Φst=0.1900,Gst=0.1339),但居群间的变异程度远远小于居群内的变异程度,居群间的基因流较大。重金属胁迫对芒萁居群的遗传分化有较大程度的影响,可能是导致芒萁适应重金属污染的分化和微进化的一个重要驱动力。     

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。     

内蒙古中东部不同草原地带羊草种群遗传分化

运用 RAPD技术对内蒙古不同草原地带分布的 5个羊草种群 (共 10 0个个体 )的遗传多样性进行分析。 31个随机引物(10 nt)在 5个羊草种群中共检测到 4 96个扩增片断 ,其中多态性片断 4 89个 ,总的多态位点百分率达 98.6 %。利用 Nei指数和Shannon指数估算了 5个种群的遗传多样性 ,并计算种群相似系数和遗传距离运用 UPGMA法进行聚类分析。结果表明 :无论是种群内还是种群间 ,羊草均存在较高的遗传变异 ,大部分的遗传变异存在于种群内 (Nei指数和 Shannon指数估算结果分别为 85 .4 %和 72 .5 % ) ,只有少部分的遗传变异存在于种群间 ;不同种群的遗传多样性存在差异 ,各种群的遗传多样性与其所处的地理位置具有显著的相关性 ;5个种群的平均遗传距离为 0 .5 0 95 ,变异范围为 0 .4 6 84～ 0 .5 4 76 ;聚类分析结果显示地理距离较近的种群遗传距离较小 ,首先聚在一起 ,而地理距离较远的种群遗传距离较大 ,说明羊草种群间的遗传分化与地理距离存在一定相关性 ;但地理距离最近的两个种群并未最先聚集 ,说明羊草种群间的分化还与其生境的异质性有关     

The effects of heavy metal pollution on genetic diversity in zinc/cadmium hyperaccumulator Sedum alfredii populations

The genetic diversity and population structure of seven populations of Sedum alfredii growing in lead/zinc (Pb/Zn) mine spoils or in uncontaminated soils from eastern and southern China were investigated using random amplified polymorphic DNA (RAPD) technology. Four of the sampled sites were heavily contaminated with heavy metals (Zn, Cd, Pb), and extremely high concentrations of Zn, Cd, and Pb were found among these corresponding populations. A significant reduction of genetic diversity was detected in the mining populations. The reduction of genetic diversity could be derived from a bottleneck effect and might also be attributed to the prevalence of vegetative reproduction of the mining populations. Analysis of molecular variance (AMOVA) and the unweighted pair group method with arithmetic mean (UPGMA) tree derived from genetic distances further corroborated that the genetic differentiation between mine populations and uncontaminated populations was significant. Polymorphism with the heavy metal accumulation capability of S. alfredii probably due to the genetic variation among populations and heavy metal contamination could have more impact on the genetic diversity and population structure of S. alfredii populations than geographic distance.     

Patterns of rapd markers and heavy metal concentrations in Perna viridis (L.), collected from metal-contaminated and uncontaminated coastal waters: are they correlated with each other?

Genetic variation due to heavy metal contamination has always been an interesting topic of study. Because of the numerous contaminants being found in coastal and intertidal waters, there is always much discussion and argument as to which contaminant(s) caused the variations in the genetic structures of biomonitors. This study used a Single Primer Amplification Reaction (SPAR) technique namely Random Amplified Polymorphic DNA (RAPD) to determine the genetic diversity of the populations of the green-lipped mussel Perna viridis collected from a metal-contaminated site at Kg. Pasir Puteh and those from four relatively' uncontaminated sites (reference sites). Heavy metal levels (Cd, Cu, Pb and Zn) were also measured in the soft tissues and byssus of the mussels from all the sites. Cluster analyses employing UPGMA done based on the RAPD makers grouped the populations into two major clusters; the Bagan Tiang, Pantai Lido, Pontian and Kg. Pasir Puteh populations were in one cluster, while the Sg. Belungkor population clustered by itself. This indicated that the genetic diversity based on bands resulting from the use of all four RAPD primers on P. viridis did not indicate its potential use as a biomarker of heavy metal pollution in coastal waters. However, based on a correlation analysis between a particular metal and a band resulting from a specific RAPD primer revealed some significant (P < 0.01) correlations between the primers and the heavy metal concentrations in the byssus and soft tissues. Thus, the correlation between a particular metal and the bands resulting from the use of a specific RAPD primer on P. viridis could be used as biomonitoring tool of heavy metal pollution.     

鼠尾藓不同居群间形态及RAPD 分析

选取采自黑龙江、内蒙古、陕西以及浙江地区的8个居群的鼠尾藓为实验材料,对不同居群的鼠尾藓叶片的形态、叶细胞的大小及植物体的形态进行了观察比较,同时运用随机扩增多态性DNA（RAPD）探讨了鼠尾藓的遗传多样性。利用13条随机引物共获得104个条带,其中多态性条带占84．62％。鼠尾藓各居群间的Dice遗传距离为0．37～0．66。POPGENE32软件分析得到种的Nei基因多样性指数（H）为0．3326,Shannon指数（I）为0．4877,遗传分化系数（GST）为0．3303。形态学观察的结果表明,鼠尾藓的植物体及叶片的形态在居群间变异较小,很好地代表了这个种的特征;而叶细胞的大小及叶尖长度的变异程度较大,说明这些形态特征易受环境影响,代表了种下水平的差异。不同居群的鼠尾藓无论在遗传上还是在形态上都表现出明显的多样性,说明鼠尾藓具有较强的适应环境的能力。     

Patterns of RAPD markers and heavy metal concentrations in <Emphasis Type="Italic">Perna viridis</Emphasis> (L.), collected from metal-contaminated and uncontaminated coastal waters: Are they correlated with each other?

Genetic variation due to heavy metal contamination has always been an interesting topic of study. Because of the numerous contaminants being found in coastal and intertidal waters, there is always much discussion and argument as to which contaminant(s) caused the variations in the genetic structures of biomonitors. This study used a Single Primer Amplification Reaction (SPAR) technique, namely Random Amplified Polymorphic DNA (RAPD), to determine the genetic diversity of the populations of the green-lipped mussel Perna viridis collected from a metal-contaminated site at Kg. Pasir Puteh and those from four relatively uncontaminated sites (reference sites). Heavy metal levels (Cd, Cu, Pb, and Zn) were also measured in the soft tissues and byssus of the mussels from all the sites. Cluster analyses employing UPGMA based on the RAPD markers grouped the populations into two major clusters; the Bagan Tiang, Pantai Lido, Pontian, and Kg. Pasir Puteh populations were in one cluster, while the Sg. Belungkor population clustered by itself. This indicated that the genetic diversity based on bands resulting from the use of all four RAPD primers on P. viridis did not indicate its potential use as a biomarker of heavy metal pollution in coastal waters. However, based on a correlation analysis between a particular metal and a band resulting from a specific RAPD primer revealed some significant (P < 0.01) correlations between the primers and the heavy metal concentrations in the byssus and soft tissues. Thus, the correlation between a particular metal and the bands resulting from the use of a specific RAPD primer on P. viridis could be used as biomonitoring tool of heavy metal pollution. The text was submitted by the authors in English.     

Molecular and cytological analyses of Deschampsia cespitosa populations from Northern Ontario (Canada).

Deschampsia cespitosa is widely dispersed around the globe, particularly in the northern hemisphere. A high tolerance to adverse environmental conditions allows D. cespitosa to colonize and dominate plots of land that are uninhabitable by other plants. The main objective of the present study was to determine the degree of genetic variation and relatedness among D. cespitosa populations from heavy metal contaminated sites and uncontaminated sites in Northern Ontario, using RAPD markers. Genomic DNA samples from individual plants were analyzed using 35 oligonucleotides of random sequence. Twenty-eight of these primers allowed amplification of random polymorphic DNA (RAPD) loci. Overall, 90% of RAPD bands were polymorphic. Analysis of molecular variance revealed that 72% of the variation could be attributed to individual differences within each of the populations. The within- and among-region variations accounted for 14 and 15% of the total molecular variance, respectively. Population-specific RAPD markers were identified. RAPD markers specific to D. cespitosa were isolated, cloned, and characterized. Cytogenetic analysis revealed a high level of aneuploidy in all the populations from Northern Ontario, with chromosome numbers varying from 2n = 18 to 2n = 26.     

Genetic structure of Suillus luteus populations in heavy metal polluted and nonpolluted habitats

The genetic structure of populations of the ectomycorrhizal basidiomycete Suillus luteus in heavy metal polluted and nonpolluted areas was studied. Sporocarps were collected at nine different locations and genotyped at four microsatellite loci. Six of the sampling sites were severely contaminated with heavy metals and were dominated by heavy metal-tolerant individuals. Considerable genetic diversity was found within the geographical subpopulations, but no reduction of the genetic diversity, current or historic, was observed in subpopulations inhabiting polluted soils. The genetic differentiation between the geographical subpopulations was low, and no evidence for clustering of subpopulations from polluted soils vs. subpopulations from nonpolluted soils was found. These results indicate that heavy metal pollution has a limited effect on the genetic structure of S. luteus populations, and this may be due to the high frequency of sexual reproduction and extensive gene flow in S. luteus, which allows rapid evolution of the tolerance trait while maintaining high levels of genetic diversity.     

陕西大豆资源遗传多样性及变异特点研究

利用42个PAPD引物对75份陕西大豆种质进行遗传多样性分析,共扩增出310个条带,平均每个引物扩增7.3个条带,多态性比率为96%;田间试验考察了13个农艺性状。陕西大豆的遗传多样性在秦岭南、北两个地区有所不同,秦岭北品种遗传多样性指数较高的性状数目和性状遗传多样性指数都大于秦岭南品种,RAPD分子标记遗传多样性指数也是秦岭北品种大于秦岭南品种,但秦岭南品种RAPD分子标记的遗传多样性指数较高的个数大于秦岭北品种。聚类分析将参试大豆材料分为三大类,基本上反映了材料的地理来源。主成分分析结果显示,前两个主成分反映了10.95%的遗传变异,基于前两个主成分值的二维散点图可以将两个地区的材料基本区分开来。AMOVA分析显示,陕西大豆品种个体间的遗传变异占总变异的92.06%,地区间的遗传变异占总变异的7.94%,二者都达到了极显著水平。研究结果表明,陕西大豆资源存在丰富的遗传多样性,秦岭北品种遗传多样性较高,但秦岭南品种有着广泛的微小变异。     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D.dyeriana.Based on random amplified polymorphic DNA (RAPD) markers,a comparative study of the genetic diversity and genetic structure of Dipteronia was performed.In total,128 and 103 loci were detected in 17 D.sinensis populations and 4 D.dyeriana populations,respectively,using 18 random primers.These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%,respectively,indicating that the genetic diversity of D.sinensis was higher than that of D.dyeriana.Analysis,based on similarity coefficients,Shannon diversity index and Nei gene diversity index,also confirmed this result.AMOVA analysis demonstrated that the genetic variation of D.sinensis within and among populations accounted for 56.89% and 43.11% of the total variation,respectively,and that of D.dyeriana was 57.86% and 42.14%,respectively.The Shannon diversity index and Nei gene diversity index showed similar results.The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high.Analysis of the genetic distance among populations also supported this conclusion.Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon.The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D.sinensis (p＜0.01),while no significant correlation was found between genetic and geographical distances among populations of D.dyeriana.This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale.This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges.We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats.Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination,and to help conserve genetic diversity.     

Genetic diversity of Mexican brook lamprey <Emphasis Type="Italic">Lampetra (Tetrapleurodon) geminis</Emphasis> (Alvarez del Villar, 1966)

Lampreys are the only surviving representatives of the oldest known vertebrates. The Mexican lamprey L. geminis (nonparasitic), is particularly interesting, because it is an endemic, biogeographical relict, and a threatened species. RAPD markers were used to describe genetic diversity in L. geminis A total of 77 specimens were collected from five populations, three in the R'o Grande de Morelia-Cuitzeo basin and two in the R'o Duero-Lerma-Chapala basin, Mexico. Eighty-eight RAPD markers were obtained from eight primers. Genetic diversity within each population was estimated using Shannon's index (S), heterozygosity (H) and gene diversity (h). These estimates revealed significant variation within populations, although a variance homogeneity test (HOMOVA) showed no significant differences among populations or between basins. Nei genetic distance values indicate a low genetic differentiation among populations. Analysis of molecular variance (AMOVA) indicates that most of the genetic diversity occurs within populations (91.4%), but that a statistically significant amount is found among populations (P0.001). Principal coordinates and cluster analyses of RAPD phenotypes show that specimens are not grouped by geographical origin. The genetic diversity found within L. geminispopulations may be explained by its breeding system and an overlapping of generations. The scarce genetic differentiation among populations is likely to the low rate of DNA change that characterizes the lamprey group.     

苹果属山荆子遗传多样性的RAPD分析

采用RAPD分子标记对东北、华北地区山荆子8个天然居群的137株个体进行了遗传多样性研究,10个引物共得到72个扩增位点,其中多态性位点63个.多态位点百分率为87.50%,有效等位基因数(Ne)为1.602 1,Nei's基因多样性(H)为0.338 6,Shannon信息指数(J)为0.496 1,表明山荆子遗传多样性水平较高.基因流(Nm)为1.735 3,说明山荆子各居群间存在一定的基因交流.居群间基因分化系数(Gst)值为0.223 7,说明虽然山荆子居群的遗传变异主要存在于居群内,但各居群间也存在着较高的遗传分化.