Investigating the genetic diversity of several varieties of Iranian tomato (Solanum lycopersicum) using RAPD and ISSR molecular markers

Background:Numerous studies have demonstrated the existence of approximately 7,500 genetic tomato varieties worldwide.Hence,it is crucial to assess the genetic diversity among tomato cultivars.This study aimed to investigate the genetic diversity of selected Iranian tomato cultivars(Solanum lycopersicum)using RAPD and ISSR molecular markers.Method:Ten RAPD primers and ten ISSR primers were employed to assess the genetic diversity among 10 tomato cultivars:Matin,RFT 112,Hirad,Golsar,Raha,Hengam,Hedah,Fasa,JS12,and Emerald.Data analysis involved the UPGMA algorithm and NTYSYSpc software.Results:RAPD analysis revealed close genetic proximity between Fasa and JS12,as well as between Raha and Hadieh.Conversely,the RFT 112,Hengam,Hirad,and Emerald cultivars exhibited significant genetic diversity within this group.ISSR primer analysis identified Hengam as the most diverse variety,while Matin,Emerald,and Vibrid,as well as Raha and JS12,displayed genetic similarities with minimal observed diversity.Furthermore,the overall analysis of the cultivars using RAPD and ISSR markers indicated that Hengam exhibited the highest diversity among all the varieties.Notably,Raha and JS12 demonstrated limited diversity in this analysis.Conclusion:This research demonstrates substantial genetic diversity among the investigated tomato varieties,with Hengam displaying the highest diversity within this group.Furthermore,ISSR markers proved more effective in determining genetic diversity in tomato plants.

Genetic variation of natural and cultured stocks of Paralichthys olivaceus by allozyme and RAPD

Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were po- lymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P. olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equi- librium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8–8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an ob- viously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect ge- netic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.

野生和养殖牙鲆(Paralichthys olivaceus)遗传差异的RAPD和ISSR研究

利用RAPD和ISSR两种分子标记技术,分析了山东威海近海野生和养殖牙鲆的遗传多样性差异。结果表明,用10个RAPD引物对两个样本各30个个体的基因组DNA进行扩增,共获得110个重复性好且带谱清晰的扩增位点,片段大小在200—2500bp之间,野生样本和养殖样本的多态位点比例分别为59.09%和60·91%,Shannon多样性值分别为16·563和16·917,野生样本内平均遗传距离为0·17427,养殖样本为0·17553,样本间遗传距离为0·17419。用6个ISSR引物共在两样本中检测到161个位点,野生样本有111个(68·94%)为多态位点,养殖样本有112个(69·57%)为多态位点,野生和养殖样本的样本内平均遗传距离分别为0·19996和0·20007,样本间遗传距离为0·20002,Shannon多样性值分别为29·254和29·688。平均位点多态率显著性检验标明,无论是利用RAPD还是ISSR技术,两群体间平均位点多态率皆差异不显著,说明第一代养殖群体的遗传结构尚未发生明显变化。

秦皇岛近海增殖放流区牙鲆(Paralichthys olivaceus)回捕群体的遗传多样性分析

本研究利用12对微卫星标记对秦皇岛近海增殖放流区内的牙鲆(Paralichthys olivaceus)回捕群体的遗传多样性进行分析,并与放流用牙鲆亲本群体和放流前群体的遗传多样性进行对比。结果显示,在3个群体中,等位基因数(N_a)为11.917–22.167个,平均观测杂合度(H_o)为0.800–0.836,平均期望杂合度(H_e)为0.814–0.845,平均多态信息含量(PIC)为0.775–0.818。其中,回捕群体的平均N_a最多,为22.167,放流前群体的平均N_a最少,为11.917。平均H_o最大的是回捕群体(0.836),最小的是放流前群体(0.800)。平均H_e最大的是亲本群体(0.845),最小的是放流前群体(0.814)。在36个群体–位点组合中,有23个群体–位点组合显著偏离哈迪-温伯格平衡状态(P<0.05)。3个群体的基因分化系数G_(st)值为0.005–0.043,遗传分化均为较弱。研究表明,3个群体均保持较高的遗传多样性,放流人工培育苗种对回捕群体的遗传多样性和遗传结构未产生明显的影响。

Assessing genetic diversity of wild populations of Japanese flounderusing AFLP markers

Amplified fragment length polymorphism （AFLP） analysis was used to evaluate the genetic diversity of four wild geographical populations of Japanese flounder （Paralichthys olivaceus）. A total of 775 loci （58.32% of which was polymorphic） in the range between 100 and 1 300 base pairs were detected from 110 individuals using seven primer combinations. The percentage of polymorphic loci detected by single primer combination for each population was calculated, ranging from 19.59% to 53.33%. Genetic similarities within and among the populations were calculated from the binary matrices of presence - absence. Phylogenetic tree of four populations was constructed by using the UPGMA method using PHYLIP Version 3.5. According to intrapopulation genetic similarities, CW population displayed the highest genetic diversity value and KY population had the lowest genetic diversity value. The distance between CW and CF populations was the farthest, which was possibly resulted from the farthest distance of Weihai of Shandong and Fujian of China compared with the geographical distance between other locations of populations. The subpopulation differentiation value （G.,） is 0. 356 5, showing a certain extent of differentiation among the four geographical populations. AFLP technology was confirmed to be an effective tool to assess within- and among-population genetic diversity of Japanese flounder. The present survey provided significant insights for research in the Japanese flounder breeding program.

Study on genetic diversity and resource conservation of amphioxus (Branchiostoma balcheri Gray) population

Amphioxus is the ancestor of vertebrates 5×108 a ago, it is a typical transitional sample of evolution from invertebrates to vertebrates. Inter simple sequence repeats (ISSRs) and random amplified polymorphic DNAs (RAPDs) technologies were applied to detect the genetic variation of 3 bulking samples and individually sampled populations in nowadays Xiamen sea areas (Xiekou, Nanxian and Huangcuo) where the amphioxuses are alive. For the bulking sampled populations, 5 ISSR and 10 RAPD primers generated 357 bands, of which 181 (50.7%) were polymorphic. Nei index and UPGMA statistical analysis indicated that amphioxuses in these 3 areas could be divided into 2 groups. The genetic distance between animals in Nanxian and Huangcuo areas was 0.07 and classified into 1 group, while the population in Xiekou belonged to another group because its genetic distances in Nanxian and Huangcuo were 0.12 and 0.14, respectively. The result was in accordance with the morphological comparison among animals from those areas. For individually sampled population, Shannon's index of genetic diversity was used to partition the diversity of the animals among these 3 sea areas, and the results showed that the indices in populations of Xiekou, Nanxian and Huangcuo, were 0.583, 0.482 and 0.374, respectively. The linear regression equation analysis for amphioxus' genetic diversity versus the environment factors revealed that granularity/sorting coefficient and water depth were the most important factors that affect amphioxus genetic diversity. On the basis of the results, the suggestions for amphioxus resource conservation in Xiamen sea areas are put forward.

Analysis of Genetic Diversity Among Sweetpotato Landraces in China

Genetic diversity of 48 sweetpotato landraces randomly sampled from Anhui,Fujian, Henan and Guangdong provinces in China was analyzed using RAPD, ISSR and AFLP markers. Thirty RAPD primers, 14 ISSR primers and 9 AFLP primer pairs generated 227, 249 and 260 polymorphic bands, respectively. AFLP markers were better than RAPD and ISSR markers in terms of the number of polymorphic bands detected and the experimental stability. These three molecular markers revealed the similar results that Chinese landraces exhibited a high level of genetic diversity, and the genetic variation of Guangdong landraces was significantly higher than those of the landraces from the other three regions. These results supported the hypothesis that China was a secondary center of sweetpotato diversity. The present results also supported the view that sweetpotato was first introduced to Guangdong and from there spread to other regions of China. The dendrogram based on the combined RAPD, ISSR and AFLP dataset could separate the 48 landraces into two groups: One mainly including 8 landraces from Guangdong and the other consisting of the remaining landraces from Guangdong and landraces from the other three regions. Thus, the utilization of Guangdong landraces should be specially considered in sweetpotato breeding.

基于ISSR和RAPD分子标记的15株糙皮侧耳遗传多样性分析

平菇(糙皮侧耳)是我国食用菌栽培主要品种之一,国内各地栽培品种较多,平菇资源的遗传多样性较丰富。为了明确平菇菌株间的亲缘关系及遗传距离,借助ISSR和RAPD两种分子标记技术,对收集到的国内主产区的15个平菇菌株进行DNA遗传多样性分析,为后期平菇适应性试验提供理论依据。通过ISSR和RAPD两种分子标记引物筛选试验,获得9对多肽引物,利用9对引物对15份平菇DNA样品进行多样性PCR扩增,并采用NTSYS-PC分析软件对供试菌株的遗传多样性进行聚类分析。结果显示,参试平菇菌株遗传距离在0.26~0.56;9对引物共扩增产生了102条清晰条带,平均每对引物扩增产生11.3条带;多态性条带90条,多态性条带比例为88%,平均每对引物组合扩增产生10条多态性条带;在遗传距离0.36的水平上,供试的15个平菇品种分为4个类群,P101和P121分别单独分为一个类群,P102、P112、P120、P122、P104、P125、P107、P113、P124归为一个类群,P117、P123、P118、P119归为一个类群。供试材料之间遗传差异大,遗传多样性较丰富,研究结果为本地区平菇产业发展及育种提供新的菌株。

RAPD和ISSR标记检测黄麻属遗传多样性的比较研究

应用RAPD和ISSR标记,分别对来自非洲地区和中国的15份黄麻野生种(10个种),以及来自中国、印度、越南、日本等地的12份黄麻栽培品种基因组DNA的遗传多样性进行检测。结果表明:(1)参与分析的所有RAPD和ISSR引物都对DNA模板浓度的梯度变化不敏感,对比RAPD和ISSR在PCR反应中的稳定性,ISSR优于RAPD;(2)25个RAPD和ISSR引物分别扩增出329条和283条带,多态比率分别为89.36%和92.5%,显然,ISSR检测出的多态性条带的能力优于RAPD;(3)RAPD和ISSR标记检测同组供试材料种间或种内的遗传相似性系数(GS)范围,分别为0.48～0.98和0.33～0.97,ISSR检测出种间遗传多样性的分辨力较RAPD为高,但两者GS相关系数高达0.955;(4)RAPD和ISSR标记的分子聚类获得了趋势相近,但不完全相同的聚类树,两种标记均能准确地把原始黄麻野生种与栽培种中的长果种和圆果种及其近缘野生种聚在不同的种群中,分子分类结果与传统经典分类相符,并揭示出种间或种内基因型的遗传差异与亲缘关系,而ISSR比RAPD能检测到种间更高的遗传差异性;(5)两种标记均可揭示种间与种内的遗传多样性及其进化的亲缘关系,可为进一步开展黄麻分子辅助育种和起源与进化研究提供有价值的理论依据。

正品龙胆遗传多样性的RAPD及ISSR分析

目的用RAPD和ISSR方法分析了4种正品龙胆的亲缘关系及遗传多样性,为正品龙胆的品种鉴定及栽培育种提供了分子生物学依据。方法优化RAPD及ISSR的PCR反应体系,对产物琼脂糖凝胶电泳结果进行数据统计。结果从100个RAPD引物和32个ISSR引物中分别筛选出10个RAPD引物和4个ISSR引物。通过遗传距离系数UPGMA聚类法分析数据,构建类聚关系树系图。结论RAPD及ISSR的PCR反应体系可以获得理想的实验结果,适用于龙胆品种遗传多样性及亲缘关系的分析。

辣椒种质遗传多样性的RAPD和ISSR及其表型数据分析

用RAPDI、SSR分子标记及28个表型性状数据对辣椒属5个栽培种的13份材料进行了分析,结果表明:23条RAPD引物共扩增出209条带,平均每个引物扩增出9.09条,多态性位点比率为83.73%;16条ISSR引物共扩增出94条带,平均每个引物扩增出5.88条,多态性位点比率为79.79%.与RAPD相比,ISSR标记检测到的有效等位基因数(Ne)及Shannon多样性指数(I)、遗传离散度(Ht)和遗传分化系数(Gst)等遗传多样性参数都较大,多态性位点比例在亲缘关系较近的一年生辣椒(Capsicum annuum)种内较高,说明ISSR有更高的多态性检测效率,并且适合亲缘关系较近的种群间遗传多样性分析.基于RAPDI、SSR的聚类与基于表型数据的聚类之间存在极显著正相关,且都能将C.annuum与其它栽培种区分开来.

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ISSR51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G5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。

用RAPD和ISSR检测的橡胶树野生种质和栽培品种的遗传多样性(英文)

用19个RAPD引物和12个ISSR引物对14份野生橡胶树种质和我国的37份栽培品种进行了遗传多样性分析.RA PD 引物共产生132条带, 多态性带占88.6% ,相似系数变化范围在0.432-0.947,ISSR引物共产生101条带,多态性带占87.1% ,相似系数为0.505-0.941。平均基因杂合度分析表明野生种质比栽培品种具有较高的遗传多样性。根据UPGM A 法对51份材料进行聚类分析,结果表明,ISSR分析中所有材料可分为2类:第一类为野生种质,第二类为栽培品种;而RAPD分析中野生种质和栽培品种不能被分为明显的两大类。虽然ISSR和RAPD的聚类分析结果存在差异,但对两种方法进行的相关分析表明,他们之间仍存在极显著相关性,相关系数为0.574。品种PR107、热研217等一些栽培品种可以通过特异带在51份供试材料中被区分开。这些结果可以对橡胶树的育种工作起到一定的指导作用,同时RAPD和ISSR 技术也是进行橡胶树品种鉴定和遗传多样性研究的有效手段。

丹江口水库翘嘴红鲌(Erythroculter ilishaeformis)遗传多样性的RAPD和ISSR分析

利用RAPD和ISSR两种分子标记技术,分析了丹江口水库野生翘嘴红鲌30个个体的遗传多样性。用10个RAPD引物对其基因组DNA进行扩增,共获得66个重复性好且谱带清晰的扩增位点,片段大小在200—2000bp之间,其中多态性位点24个,多态位点比例为36.36%;个体间遗传距离在0.0338—0.1426之间,平均为0.0941;Shannon信息指数为0.1083。用10个ISSR引物共检测到73个位点,其中多态性位点28个,多态位点比例38.36%;个体间遗传距离在0.0376—0.1652之间,平均为0.1058;Shannon信息指数为0.1196。结果显示,丹江口水库野生翘嘴红鲌的多态位点比例、平均遗传距离和Shannon信息指数均较大,说明其有较丰富的遗传多样性。

云南黑籽南瓜种质遗传多样性的RAPD和ISSR分析

采用RAPD和ISSR分子标记技术对来源于云南省6个地州13份黑籽南瓜种质进行遗传多样性分析。结果表明:6个RAPD和6个ISSR引物分别扩增出43条和41条带,多态性比率分别为90.70%和51.21%;RAPD和ISSR标记检测供试材料的遗传相似性系数范围分别为0.340～0.895和0.162～0.941I,SSR检测多态性效果高于RAPD。RAPD标记聚类分析将供试种质分为3个类群5组;ISSR标记聚类分析将供试种质分为4个类群6组,RAPD和ISSR标记的遗传相似性系数呈显著相关。基于UPGMA聚类结果,可为黑籽南瓜的引种栽培或品种改良提供参考。

利用RAPD和ISSR分子标记分析地黄种质遗传多样性

用RAPD与ISSR技术对地黄的 8个品种和 2个脱毒品系进行了种质遗传多样性分析。分别从 80条RAPD引物和 4 4条ISSR引物中筛选出适合地黄种质分析的 17条RAPD引物和 10条ISSR引物用于RAPD和ISSR分析。 17条RAPD引物共扩增出 177条带 ,多态性位点数为 10 9;多态性位点比率为 6 1 5 8% ;平均多样性指数 (I)为0 3135 ;每个位点的有效等位基因数 (Ne)是 1 36 4 1;10条ISSR引物共扩增出 110条带 .多态性位点数为 79;多态性位点比率为 71 5 8% ;平均多样性指数 (I)为 0 35 77;每个位点的有效等位基因数 (Ne)是 1 4 0 37。基于扩增条带数据库建立了各自的Jaccard遗传相关系数矩阵 ,构建了相似的分子树状图 ,将 10个供试材料分为 2类 :一类群含组培 85 5、大田 85 5、组培 930 2、大田 930 2、金状元和金白 6个材料 ;另一类群含北京 1号、大红袍、地黄 910 4和野生地黄 4个材料。两种分子标记的分析结果呈极显著正相关 (r=0 6 4 9)。结果表明 ,RAPD与ISSR标记适合于地黄种质遗传多样性分析 。

野生地黄种内遗传多样性的RAPD，ISSR分析

目的：分析野生地黄群落之间的遗传差异类型，比较RAPD和ISSR分子标记在分析地黄野生种质遗传多样性中的应用。方法：应用RAPD和ISSR分子标记技术，对55份地黄材料进行了遗传差异分析。结果：平均每条RAPD引物扩增出16．00条带，平均每条ISSR引物扩增出19．08条带，多态性位点百分率分别为89．58％和94．32％；聚类分析表明，55份地黄分成7大类群。结论：野生地黄具有丰富的遗传多样性；不同地理居群的相似性为0．63—0．98；ISSR标记比RAPD标记能检测到地黄种内更高的遗传差异性。

山楂(Crataegus pinnatifida Bge．)遗传多样性的RAPD和ISSR标记分析

利用RAPD和ISSR标记对35份山楂（Crataegus pinnatifida Bge．）资源进行了DNA多态性分析。12个RAPD引物共扩增出110条清晰的谱带，其中89条显示多态性，平均每个引物扩增出7．4条多态性谱带。13个ISSR引物共扩增出110条清晰的谱带，其中94条显示多态性，平均每个引物扩增出7．2条多态性谱带。基于RAPD和ISSR标记，利用UPGMA分别构建了35份山楂资源的聚类树状图。距离系数分别为0～0．62（RAPD）和0～0．64（ISSR），表明山楂具有较高的遗传多样性。

鸢尾属植物遗传多样性的RAPD和ISSR分析

应用RAPD和ISSR标记技术,对来自不同产地的鸢尾属(IrisL.)4个野生种的遗传多样性进行了分析。结果表明,12个RAPD和ISSR引物分别扩增出225和196条带,多态性条带分别为215和196条,多态性条带百分率分别为95.56%和100.00%;种间总基因多样度分别为0.368 9和0.357 5、种内基因多样度分别为0.107 7和0.138 0,表明鸢尾属种间遗传多样性较高,且种间变异大于种内变异。4个野生种中,蝴蝶花(I.japonicaThunb.)的遗传组成较为丰富。此外,种内遗传关系与地理分布和环境差异有一定的相关性。

基于ISSR和RAPD标记的四个夏蜡梅种群的遗传多样性研究

利用ISSR和RAPD技术对4个夏蜡梅种群25个单株的遗传多样性进行研究。从60条简单重复序列引物中筛选出了10条引物在25个个体中共检测到62个可重复的位点,其中多态位点为41个,总的多态位点百分率为65.60%;从60条寡居核苷酸引物中筛选出了10条引物共扩增出52个位点,其中多态性位点31个,多态性位点的百分率为57.50%。Shannon指数估算的夏蜡梅群体总遗传多样性为0.3737,各群体平均遗传多样性为0.1645。Nei’s指数估算的夏蜡梅群体总遗传多样性为0.2528,各群体平均遗传多样性为0.1117;物种水平的有效等位基因数为1.4473,平均值为1.1972。4个自然群体的基因分化系数Gst=0.5297,即总的变异中有52.97%的变异存在于群体间,而群体内的遗传变异占总变异的47.03%,群体间遗传距离平均值0.2187。也表明了夏蜡梅4个群体间出现了遗传分化。夏蜡梅群体间的基因流较低,Nm=0.4450。聚类分析将4个种群聚为2支,且聚类结果表现出明显的地域性特征。作为狭域分布种,夏蜡梅各群体间存在遗传分化和多样性程度不高,对夏蜡梅种群多样性的就地和迁地保护势在必行。