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。     

木霉肽类代谢产物对豇豆土著根瘤菌遗传性状的影响

利用RAPD与ISSR分子标记检测手段,分析了哈茨木霉T2-16肽类代谢产物处理豇豆土著根瘤菌,对其遗传性状的影响,同时,比较了RAPD和ISSR两种不同分子标记在检测根瘤菌种间的遗传相似性以及遗传变异性的分辨力.实验中,从100条引物中筛选到具有多态性的ISSR引物5条,从80条引物中筛选到具有多态性的RAPD引物6条,用5条ISSR引物扩增出54条带,多态性条带比率为75.93 %;6条RAPD引物扩增出61条带,多态性条带比率为68.85 %.两种分子标记均能揭示出处理前后根瘤菌间的遗传差异,但ISSR标记比RAPD标记可检测到更大的遗传变异.根据两种标记的结果,对供试的根瘤菌进行聚类分析,结果表明,土著根瘤菌经木霉肽类代谢产物处理后,与出发菌株相比,表现出一定程度的遗传分化和遗传差异性.     

蝴蝶兰种质资源遗传多样性的ISSR分析

利用ISSR分子标记对24个蝴蝶兰(Phalaenopsis)栽培品种进行了遗传多样性分析.筛选出的12个ISSR引物共扩增出301条清晰的谱带,其中多态性条带268条,多态性比率为89.1%.品种间的遗传相似系数在0.37～0.93之间,表明品种间具有较高的遗传多样性.UPGMA聚类分析可将供试材料分为3组,分类结果与材料来源及花器官表型具有密切的关系.     

特有珍稀濒危植物景东报春遗传多样性的ISSR分析

景东报春(Primula interjacens Chen)是一分布在中国云南中南部的稀有的、分布区狭窄的特有种.该种含2个变种:原变种景东报春(P.interjacens Chen var.interjacens),现存一个居群;和光叶景东报春(P.interjacens Chen var.epilosa C.M.Hu),有2个居群.本文用ISSR分子标记对景东报春现存的3个居群进行了遗传多样性分析.和我们最初的预计相反,尽管居群小、分布区狭窄,景东报春的遗传多样性却很高(在居群水平的多态位点率P=59.75%,基因多样性HE=0.236 8,Hpop=0.345 9;在种的水平的多态位点率P=75.47%,遗传多样性HT=0.320 5,总的遗传多样性Hsp=0.4618).推测异型花和杂交可能是维持高的遗传多样性的原因.Nei的遗传多样性水平(26.13%)和Shannon多样性指数(25.09%)显示居群间存在中度分化.尽管景东报春原变种和变种在形态上有区别,但用非加权配对算数平均法(UPGMA)聚类分析的结果表明,两者并没有明显的遗传分化,可处理为一个单一的分类单元.基于观察到的居群遗传信息,建议采取就地保护和迁地保护的保护措施.     

利用ISSR揭示不同类型月季遗传多样性

本研究应用正交设计法对ISSR反应体系中的各个主要影响因子进行了优化筛选,确立了适合月季ISSR-PCR反应的最佳体系.结果表明,25μL的ISSR反应体系中各组分的最适浓度分别为:1×PCR缓冲液、1 U Taq DNA聚合酶、800 pmol/L引物、0.16 mmol/L dNTPs、Mg2+ 1.5 mmol/L.筛选了33个ISSR引物,共得到了11个多态性比较高的ISSR引物,占所筛引物的33.33%.利用筛选出的11条ISSR引物对3种月季类型的23份月季材料进行遗传多样性分析,共扩增出477条DNA带,其中多态性位点有14个,平均每条引物可以检测到4.5个多态性位点.用NTSYS软件对样品进行了UPGMA聚类分析,聚类结果表明丰花月季基本能聚为一类,切花月季与藤本月季交叉聚在一起.这表明月季种质的遗传差异与其应用分类的相关性不紧密.     

特有珍稀濒危植物景东报春遗传多样性的ISSR分析(英文)

景东报春(Primula interjacens Chen)是一分布在中国云南中南部的稀有的、分布区狭窄的特有种。该种含2个变种:原变种景东报春(P.interjacens Chen var.interjacens),现存一个居群;和光叶景东报春(P.interjacens Chen var.epilosa C.M.Hu),有2个居群。本文用ISSR分子标记对景东报春现存的3个居群进行了遗传多样性分析。和我们最初的预计相反,尽管居群小、分布区狭窄,景东报春的遗传多样性却很高(在居群水平的多态位点率P=59.75％,基因多样性H_E=0.2368,Hpop=0.3459:在种的水平的多态位点率P=75.47％,遗传多样性H_T=0.3205,总的遗传多样性Hsp=0.4618)。推测异型花和杂交可能是维持高的遗传多样性的原因。Nei的遗传多样性水平(26.13％)和Shannon多样性指数(25.09％)显示居群间存在中度分化。尽管景东报春原变种和变种在形态上有区别,但用非加权配对算数平均法(UPGMA)聚类分析的结果表明,两者并没有明显的遗传分化,可处理为一个单一的分类单元。基于观察到的居群遗传信息,建议采取就地保护和迁地保护的保护措施。     

栽培沉香遗传多样性的ISSR和AFLP分析比较

探讨2种分子标记技术在沉香属药用植物遗传多样性研究中的应用。用ISSR和AFLP分子标记分析了海南、云南、广东、广西等地17份沉香属植物的遗传多样性。14个ISSR引物、8对AFLP引物分别检测到119、919个位点,多态位点百分率分别为73.95%、86.94%。由于AFLP标记具有较高的多态性位点检测效率,AFLP标记分析的遗传多样性参数高于ISSR。虽然基于Nei’s遗传距离的聚类分析结果存在着一定的差异,但用Mantel检测对两种方法检测的遗传一致度进行相关性分析表明,它们之间存在着明显的相关性（r=0.7705,P=0.0003）。ISSR标记与AFLP标记均能应用于沉香属植物的遗传多样性研究。两种标记的研究结果均揭示出沉香属植物具有较高的遗传多样水平。     

苦瓜种质遗传多样性的RAPD和ISSR分析

采用RAPD和ISSR分子标记技术对38份苦瓜种质进行遗传多样性分析。结果表明:10个RAPD和10个ISSR引物分别扩增出93条和81条带,多态性比率分别为50.54%和61.29%;RAPD和ISSR标记检测供试材料的遗传相似性系数(GS)范围,分别为0.287~1和0.221~1,ISSR(平均GS值0.672)检测多态性效果高于RAPD(平均GS值0.694)。RAPD标记聚类分析将供试种质分为3个类群6组,分类结果与苦瓜瓜瘤的表型分类比较相似;ISSR标记聚类分析将供试种质分为3个类群7组,ISSR标记划分类群与形态上以颜色分类比较接近。RAPD和ISSR标记的遗传相似性系数呈显著相关(r=0.550)。两个标记整合后聚类分析可检测到更大的遗传变异,结果与苦瓜的农艺性状分类和地理分布有一定的相关性。     

云南普通野生稻遗传多样性和亲缘关系

野生稻（Oryza rufipogon）是稻属的重要组成部分,具有许多优良性状,是水稻遗传改良的天然基因库。本研究通过对形态学性状的观测,及ISSR和RAPDUPGMA聚类分析,将云南普通野生稻划分为4个类型,即元江类型、景洪紫杆直立型、景洪绿杆直立型和景洪匍匐型。在供试材料中筛选到具有多态性的ISSR和RAPD引物各11个,ISSR引物扩增出多态带113条,多态性条带比率（PPB）为82．26％,RAPD引物共扩增出多态性条带76条,PPB值为76．77％,两种分子标记的分析结果呈极显著正相关（r=0.951）。此外UPGMA聚类结果表明,云南普通野生稻不同类型与其它地区普通野生稻之间的遗传亲缘关系差异明显。     

利用RAPD和ISSR标记分析烤烟品种间遗传关系

利用RAPD和ISSR标记对22份烤烟(Nicotiana tabacumL.)品种进行了遗传关系研究。在RAPD分析中筛选到13个引物,共扩增出167条带,其中多态性带50条,多态性比率为29.9%;在ISSR分析中筛选出7个引物,共扩增出96条带,其中多态性带44条,多态性比率为45.8%。两种标记相结合估算出的品种间遗传相似系数在0.881~0.979之间,平均为0.933。单独基于RAPD标记和ISSR标记的聚类结果有一定差异;两种标记结合起来的聚类分析结果与系谱信息吻合程度更高。定向选择可能对烤烟品种间遗传关系有较大影响;国外引进品种与国内育成品种并未完全分开,表明分子水平的遗传关系和地理来源间缺乏必然联系。     

Development and characterization of microsatellite loci in Taihangia rupestris (Rosaceae), a rare cliff herb

? Premise of the study: Microsatellite primers were developed for the rare Taihangia rupestris (Rosaceae) to evaluate genetic diversity, population genetic structure, mating system, and demographic events of this species. ? Methods and Results: Ten primer sets were developed using an enriched genomic library and were successfully amplified in T. rupestris var. ciliata and T. rupestris var. rupestris. The number of alleles per locus ranged from 2 to 21; the observed and expected heterozygosities ranged from 0.300 to 0.950 and from 0.328 to 0.956, respectively, in the two varieties. ? Conclusions: The markers described here will be useful for studies of genetic variation, genetic structure, and mating systems of T. rupestris, which are important for the future conservation of this rare species.     

多叶重楼遗传多样性的RAPD分析

应用RAPD技术检测了多叶重楼(Paris polyphyfzo)2个变种4个居群的遗传多样性,并与1个凌云重楼(P．cronquistii)居群进行了比较。选择的16个随机引物在5个居群中共检测到246个多态位点。在居群水平上,滇重楼2个居群的多态位点百分比(PP鳓分别为57．43％和54．67%,Shannon指数分别为0．3080和0．2830;七叶一枝花2个居群的PPB分别为56．33％和57．75%,Shannon指数分别为0、3080和0．3293。在变种水平上,滇重楼的PPB为75．14％,Shannon指数为0．3922,遗传分化系数(Gst)为0．3085;七叶一枝花的PPB为80．31％,Shannon指数为0．3992,遗传分化系数(Gst)为0．3726;在种的水平PPB达92．05%,遗传分化系数Gst达0．5151。聚类分析显示滇重楼和七叶一枝花有较近的亲缘关系,而与凌云重楼遗传距离较远。此结果从分子水平上支持了过去将滇重楼和七叶一枝花划分为1个种下2个变种的形态分类观点。     

云南松及其近缘种的遗传变异与亲缘关系

The authors investigated the genetic diversity of 29 natural populations representing Pinus yunnanensis Franch. and its two close relatives, P. densata Mast. and P. kesiya Royle ex Gordn. var. langbianensis (A Chey.) Gaussen. Horizontal starch gel electrophoresis was performed for macrogametophytes collected from populations in Yunnan, Sichuan and Guangxi. Allozyme data for 33 loci of 14 enzymes demonstrated high levels of genetic variation at both population and species levels in comparison with other conifers, with the mean values for populations being P=0.694, A=2.0 and He=0.145 for P. yunnanensis; P=0.714, A=2.0 and He=0.174 for P. densata; and P=0.758, A=2.1 and He=0.184 for P. kesiya var. langbianensis. Based on Wright's F-statistics, the fixation index of P. yunnanensis, P. densata and P. kesiya var.langbianensis were 0.101, 0.054 and 0.143, respectively, indicating that the populations were largely under random mating. Based on Nei's genetic distance, the genetic differentiation was not obvious among the three species (i.e. the genetic distance was less than 0.075). Because of the wider distribution of P. yunnanensis with greater variety of habitats, it was shown that the genetic differentiation among the P. yunnanensis populations was larger than that of the populations of the other two species. According to morphological, geographic and allozymic evidences, the authors suggested that the three species be better treated as varieties under a single species. In addition, the extensive gene flow among the three pine species resulted in great genetic diversity and evolutionary potential. Also, high level of genetic variation of P. yunnanensis provides important basis for its genetic improvement and breeding in future.     

Microsatellite variation of cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis> Crantz) in home gardens of Chibchan Amerindians from Costa Rica

We used 13 microsatellite marker loci to determine the genetic diversity of cassava (Manihot esculenta Crantz) grown in home gardens in two Chibchan Amerindian reserves in Costa Rica. We compared the levels of genetic diversity in the reserves with that of commercial varieties typically cultivated in Costa Rica. We found high levels of genetic diversity among cassava plants. Overall, 12 of the 13 loci examined were polymorphic in each Amerindian reserve (P = 92.3). Moreover, we found 36 alleles in the Coto Brus Reserve and 33 in the Talamanca Reserve. In the commercial varieties only nine loci were polymorphic (P = 69.2), and we only found 23 alleles. Heterozygosity was high for all groups of cassava (Coto Brus, Talamanca, and commercial varieties), but it was higher among the commercial varieties. The levels of heterozygosity and allele diversity indicate that there is significant genetic diversity in the home gardens that we examined. Another indication of the high diversity found in these gardens is the number of distinct multilocus genotypes, 28 at Coto Brus and 19 at Talamanca. There was also more than one distinct multilocus genotype found within the commercial varieties, as three were found in Valencia and four in Manyi. Our data also revealed low levels of genetic differentiation between the three groups of cassava (F_st = 0.03), and Nei’s genetic distances ranged from 0.0167 to 0.0343. In addition, F estimates (F_is and F_it) indicate excess heterozygotes, both at the subpopulation and the population level. A hierarchical analysis of the genetic variation revealed that variation between sampling locations within each of the three groups of cassava was larger than that between groups (Theta S = 0.0775 and Theta P = 0.0204, respectively). The variety Manyi was the group genetically most distant from all others. We discuss the consequences of these findings for in situ conservation of genetic resources.     

在韩国境内Potentilla fragarioides var .sprengeliana的遗传多样性和种群结构(英文)

根据 2 2个等位酶位点遗传变异 ,探讨了韩国境内委陵菜 (PotentillafragarioidesL .var.sprengeliana)的遗传多样性和种群结构。酶位点的多态位点百分比为 5 9 1%。种和种群水平上的遗传多样性比较高 ,分别为Hes=0 .2 10 ,Hep=0 .199;而种群的分化水平则相对较低 (GST=0 .0 74)。 19个种群中随机交配的偏差为FIS=0 .331。每代迁移数的间接估计 (Nm=3.15 )表明该种在韩国的种群间基因流较高。另外 ,固定指数分析显示在一些种群和位点有轻微的杂合子缺乏。种群间平均遗传一致度为 0 985。这些韩国委陵菜种群存在于较为均一的生境 ,这很有可能是造成其种群遗传一致性较高的原因。     

Patterns of Genetic Diversity and Population Structure of the Clonal Herb,Potentilla fragarioides var. sprengeliana (Rosaceae) in Korea

The genetic diversity and population structure of Potentilla fragarioides var. sprengeliana(Rosaceae) in Korea were investigated using genetic variation at 22 allozyme loci. The percent of polymorphic loci within the enzymes was 59.1%. The genetic diversity at the species level and at the population level was high (Hes=0.210; Hep=0.199, respectively), whereas the extent of the population divergence was relatively low (GST=0.074). FIS, a measure of the deviation from random mating within the 19 populations, was 0.331. An indirect estimate of the number of migrants per generation (Nm=3.15) indicates that gene flow is high among Korean populations of the species. In addition, analysis of fixation indices revealed a slight heterozygote deficiency in some populations and at some loci. The mean genetic identity between populations was 0.985. It is highly probable that the trend of genetic uniformity in a relatively homogeneous habitat is thought to be operated among Korean populations of P. fragarioides var.sprengeliana.     

SSR分子标记检测出的花生类型内遗传变异

花生是我国重要的食用油和蛋白质来源作物,鉴定其DNA分子多态性对品种改良和资源评价具有重要的意义。从已公布的花生Genomic-SSR和EST-SSR引物中筛选出34对引物,用来分别鉴定花生4大类型各24份共96份品种资源的分子变异,其中龙生型资源全部来自广西,普通型资源中有11份从国外引进,有13份来自广西和国内其他省市,多粒型资源只有两份来自中国,其他22份分别来自印度、美国和非洲等地,珍珠豆型资源中有22份是来自中国各地的育成品种或农家品种,有2份来自国外。研究结果为:分别有10～16对SSR引物能在4大类型花生资源中扩增出多态性DNA片段;这些多态性SSR引物都具有多位点特性;首次为SSR分子标记设立了一个新的评价指标——区别指数,多态性SSR引物的区别指数最高达0.992;资源间的平均遗传距离,多粒型为0.59,普通型为0.48,珍珠豆型为0.38,龙生型为0.17。根据遗传距离采用最长距离法对4大类型花生资源分别进行了聚类分析,构建了资源间的遗传关系图,花生4大类型可进一步分成不同类群,资源间的亲缘关系与其来源相关。观察到PM15和PMc297的扩增产物具有类型特异性,PM15能在龙生型、普通型和多粒型花生资源中扩增出多态性条带,而在珍珠豆型花生中扩增条带完全相同,PMc297也有相似的扩增结果。由于在多粒型花生资源中检测出的遗传多样性最丰富,研究结果支持西班牙专家Krapovickas 1994年公布的花生栽培种分类系统。总之在花生4大类型内资源中能检测出丰富的SSR分子标记,开发出更多的SSR分子标记将能充分揭示花生分子水平的变异,从而使花生遗传图谱构建、分子标记辅助育种成为可能。     

Clonal structure and patterns of allozyme diversity in the rare endemic Cycladenia humilis var. jonesii (Apocynaceae)

The rare endemic Cycladenia humilis var. jonesii (Jones cycladenia) has low levels of sexual reproduction. Enzyme electrophoresis was used to explore possible causes of low seed set and high fruit abortion by assessing the clonal structure and genetic diversity in populations. The seven populations studied were composed of multiple, highly interdigitated clones; thus low fruit set is not likely to be due to a scarcity of mates. Genotype frequencies did not differ significantly from Hardy-Weinberg proportions, suggesting that populations are not highly inbred. Jones cycladenia exhibited high levels of genetic diversity at both the population level (A = 1.7; P = 37; He = 0.14) and the taxon level (A = 2.7; P = 60) in comparison to other plants. These data suggest that genetic drift is unlikely to have left this taxon genetically depauperate. Furthermore, we detected little divergence among geographically disjunct populations of Jones cycladenia (FST = 0.10). In comparison, Jones cycladenia populations were highly differentiated from a population of the taxon's close relative, C. h. var. humilis (mean genetic identity = 0.76). Our study suggests that other reasons for low fruit set in Jones cycladenia, such as resource or pollinator limitation, or genetic load, should be explored in future research.