利用SSR分析小豆种质遗传多样性

摘要：小豆是一类重要的食用豆,本研究利用45对SSR引物对小豆基因组DNA进行了SSR标记的筛选鉴定,共筛选出多态性良好、扩增效果稳定的SSR引物18对。利用筛选出的18个SSR标记,分析了来自我国栽培小豆优异种质53份和日本引进种质27份,旨在阐明其遗传多样性特点,为育种利用提供理论依据。结果表明,在所有参试的80份小豆种质中共鉴定出等位变异92个,平均每个位点为5.1个;其中53份国内小豆和27份日本小豆的等位变异数分别为89个和74个,平均每个位点分别为4.9个和4.1个。所有供试小豆平均每个位点的多态信息含量（PIC）为0.64,变化范围为0.23～0.83,其中国内小豆的平均PIC值为0.63,变化范围为0.23~0.86;日本小豆平均PIC值为0.61,变化范围为0.20~0.81。国内栽培小豆和日本小豆在等位变异数、多态信息含量（PIC）、遗传相似性系数均存在差异,UPGMA聚类分析将参试的80份小豆明显分为五大类,聚类结果与小豆种质地理起源呈现出一定的相关性。试验表明,在小豆遗传育种中,可以通过种质资源相互利用来拓宽育成品种的遗传基础,同时这些SSR标记对于小豆资源DNA指纹图谱构建、遗传作图、基因型鉴定及分子标记辅助育种具有重要意义。     

不同生态区谷子品种表型鉴定及SSR遗传多样性分析

为了明确不同生态区谷子品种的遗传多样性和遗传结构,本研究对175份来源不同的谷子品种进行29个表型性状鉴定和36对简单序列重复（SSR）标记分析。结果表明,谷子数量性状的平均遗传多样性指数最高,质量性状最低;不同类性状中,单穗码数、开花至成熟日数和粒色遗传多样性指数最高,分别为2.082 8、2.012 5和1.078 7。36对SSR引物中,共检测到401个等位基因,平均11.4个,Shannon指数和多态性信息含量（PIC）平均为2.017 2和0.810 6;标记B142、B225的Shannon指数和PIC值最高,是评价谷子遗传多样性的理想SSR标记。不同生态区表型和SSR标记多样性比较结果表明,春谷中晚熟区（ML）遗传多样性指数最高,春谷早熟区（EM）次之,夏谷区（SS）最低;春谷中晚熟区谷子品种有10个性状指标最高,且全生育期最长。春谷中晚熟区不同类型材料中,汾阳的质量性状遗传多样性指数和SSR标记Shannon指数均最高,长治数量性状遗传多样性指数最高,太原Ⅱ的生育期遗传多样性指数最高;不同类型材料间差异较低,其中7个数量性状指标差异未达显著水平;汾阳的株高最高,出谷率最...     

吉富罗非鱼雌雄群体遗传差异的SSR分析

为探讨吉富罗非鱼（genetic improvement of farmed tilapia,GIFT）雌、雄群体间的遗传差异,本研究对国家级广西南宁罗非鱼良种场雌、雄吉富罗非鱼进行了遗传差异分析。研究结果表明,选取的11对SSR引物中有10对能获得稳定的目的条带;每个SSR基因座的等位基因数在2～4个之间,雌性罗非鱼的平均等位基因（Na）2.9个,稍高于雄性的2.8个;雌、雄吉富罗非鱼平均观察杂合度（HO）分别为0.4183和0.4154,多态信息含量（PIC）分别为0.4048和0.3932,属中度多态;雌雄个体间的遗传距离和相似性指数分别为0.0908和0.9132。此外,SSR基因座PRL-SO2在雄鱼中偏离Hardy Weinberg平衡（P〈0.005）。上述结果表明,吉富罗非鱼雌、雄群体的SSR多态性基本相同,推测这两者基因组间的差异较小。     

东南亚62个籼型水稻亲本SSR遗传多样性分析

为探讨东南亚水稻亲本间的遗传多样性,以东南亚62份籼型水稻亲本为材料,选择29对水稻功能基因标记以及平均分布于12条染色体、多态性较高和条带清晰的72对SSR引物进行遗传多样性和聚类分析。结果表明,32对多态性引物在62份水稻材料中检测到201个等位基因,每个位点等位基因数(Na)变异范围为2~12,平均为6.281;有效等位基因数(Ne)变异范围为1.067~5.399,平均为2.867;多态信息含量(PIC)变异范围为0.061~0.789,平均为0.515;平均Shannon信息指数(I)为1.176,变幅为0.143~1.908;观察杂合度(Ho)为0.977,范围为0.936~1.000;期望杂合度(He)为0.439,范围为0.179~0.937;固定指数(Fis)的范围为0.882~1.000;基因流(Nm)的范围为0~0.0157。聚类分析表明,在遗传相似系数为0.805处,东南亚水稻亲本被分为6大类,同一国家大多数品种聚为一类。相对于普通分子标记,功能基因标记的平均等位基因数不高,其遗传多样性参数也低于普通分子标记,东南亚各国水稻品种间的亲缘关系较近,遗传多样性程度较低。研究结果为水稻育种亲本选配、新种质资源的创制以及杂种优势利用等提供了科学依据。     

抗麦长管蚜小麦的遗传多样性及SSR标记与麦长管蚜抗性的关联分析

麦长管蚜(Sitobion avenae)是影响中国小麦(Triticum aestivum)生产的主要害虫之一。本研究选用99个简单重复序列(simple sequence repeat,SSR)标记对54份苗期和成株期麦长管蚜抗性一致的小麦品种进行聚类分析和麦长管蚜抗性关联分析。结果表明,99个SSR标记共鉴定出1 038个等位变异,平均每个位点的等位变异数为10.48个,变异数目在2~30个之间;SSR标记位点的多态性信息含量(PIC)的变异范围为0.178 6~0.973 8,平均为0.717 8。聚类分析表明,54份小麦品种聚成2大类群,大部分地理来源相近或麦长管蚜抗性相似的材料聚于同一亚类群。通过一般线性模型(general linear model,GLM)和混合线性模型(mixed linear model,MLM)两种关联分析模型,分别获得16个和4个麦长管蚜抗性相关联的标记,这些标记分别位于11条染色体的14个染色体臂上,GLM分析中各标记对表型变异解释率为0.221 9~0.556 7,MLM分析中各标记对表型变异解释率为0.029 5~0.063 3。研究结果为小麦抗麦长管蚜杂交育种及分子标记辅助育种提供理论依据。     

利用SSR标记分析部分粳稻品种的遗传多样性

以我国16个粳稻品种为材料,包括水稻雄性不育系、恢复系、杂交种以及少量常规稻,从分布在水稻基因组的12条染色体上300对SSR引物中筛选出14对多态性丰富、重复性高的引物,这些引物在16份材料间共扩增出61个多态性片段,平均每对SSR引物可检测到4.36个等位基因。聚类分析表明:品种间遗传相似系数在0.57~0.97之间。生产中应用的BT型水稻雄性不育系与恢复系分别聚类于不同的类群,遗传关系较远。     

利用简单重复序列(SSR)标记分析太湖稻区现代粳稻品种的遗传多样性

太湖地区有丰富的粳稻(Oryza satiua ssp.japonica)种植资源,随着品种大面积推广应用,育种材料的遗传基础趋窄,相似性增高,使粳稻育种突破困难。本研究利用分布于水稻(Oryza satiua L.)12条染色体上的24对简单重复序列(simple sequence repeat,SSR)引物对太湖地区的42份粳稻品种进行遗传多样性分析。结果表明,有23对SSR引物在42份粳稻材料间表现出多态性;23对引物共检测到105个等位基因,每对SSR引物检测到等位基因为2~8个,平均为4.57个,有效等位基因共有56.43个,平均每个位点为2.45个。每个多态位点的多态信息含量(polymorphism information content,PIC)变幅为0.083 0~0.8079,平均为0.496 6;每个粳稻材料多态性位点数的变幅为6~19,等位基因总数的变幅为27~55;42份粳稻品种间的遗传相似系数变幅为0.391~0.990,平均为0.610,遗传相似系数在0.50~0.80之间的材料占全部的74.45%,供试材料相似度高;非加权配对算术平均法(unweighed pair group method with arithmetic mean;UPGMA)聚类结果显示,遗传相似系数为0.50,42份粳稻材料可以分为两个类群,一个类群包含19份常规粳稻,另一个类群包括其他23份杂交粳稻。结果显示,太湖地区的粳稻品种总体上遗传背景相似度高,遗传多样性不够丰富,育种工作有待进一步加强新的基因资源引进和利用,创新水稻育种材料。本研究结果为新品种选育提供技术支持和理论根据。     

山西芝麻种质资源SSR遗传多样性及群体结构分析

为探明山西芝麻种质资源的遗传特性,本研究利用30对简单重复序列标记(SSR)对71份山西芝麻种质资源进行遗传多样性分析及群体结构分析。结果表明,30对SSR标记共检测到144个等位基因位点,平均每个SSR标记4.800个等位基因;有效等位基因数在1.058~5.149之间,平均2.805个;Shannon指数变幅为0.128~1.813,平均为1.096;Nei's遗传多样性指数变幅为0.055~0.806,平均为0.558;多态性信息含量变幅为0.053~0.783,平均为0.515。基于SSR标记对参试材料进行聚类分析,遗传相似系数为0.21~0.67,在遗传相似系数0.27处将参试材料分为6个类群;基于SSR标记对参试材料进行群体结构分析,将参试材料划分为5个组群。综上所述,山西芝麻种质资源间遗传差异相对较高,具有丰富的遗传多样性,在今后芝麻种质资源创制利用中,加大山西芝麻种质资源的开发与利用,可为芝麻品种遗传改良和优异基因发掘奠定良好的基础。     

利用SSR标记对中国柚类资源及近缘种遗传多样性研究

利用SSR标记研究了122份我国柚(CitrusgrandisOsbesk)类资源及近缘种遗传多样性。31对SSR引物从供试材料中检测出335个等位基因变异,平均每个位点可检测到9.85个等位基因。位点多态信息量(PIC)变幅为0.1939!0.9073,平均为0.7085。用UPGMA方法将122份材料分成7个组群,110个柚类品种在相似系数0.712,可细分成18个亚组,主要由沙田柚品种群、文旦柚品种群及庞大的杂种柚品种群组成。     

华南沿海地区南方夏大豆遗传多样性的SSR分析

利用60个SSR位点对来自华南地区9个省份的191份南方夏大豆（Glycine max）进行分析，结果显示：在191份材料中共检测到663个等位变异，平均每个位点的等位变异数为11．05个；Shannon-Weaver指数平均值为1．67，材料的成对相似系数范围为0．19-0．95，平均相似系数为0．29，说明华南地区南方夏大豆材料间的相似程度高，与表型性状表现相一致；在聚类分析中，以相似系数0．26为划分标准，可将夏大豆分为4个类别，但9个省（市）之间并没有明显的界限，相近纬度和同一流域夏大豆趋向于聚在一起；通过等位变异数及遗传多样性指数分析发现，华南地区南方夏大豆分子遗传多样性中心可分成2个：一个以安徽省为中心的长江流域．另一个以广西壮族自治区为中心的珠江流域。由于多数相同来源的夏大豆分别聚在不同的类别中，为充分利用本地资源拓宽育种遗传基础提供了理论依据。     

Genetic diversity comparison between Chinese and Japanese soybeans (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.) revealed by nuclear SSRs

Soybean (Glycine max (L.) Merr.) has a long planting history in both China and Japan. In order to investigate the genetic relationship between Chinese and Japanese soybeans, 205 Chinese soybean accessions, that represent the seven different soybean ecotypes, and 39 Japanese soybean accessions from various regions were analyzed by using 46 SSR loci. In total 745 alleles were detected with an average allele number of 16.2 per locus. The allelic frequencies varied from 0.002 to 0.554 with an average of 0.06. Cluster analysis with UPGMA separated the Chinese accessions from Japanese accessions, suggesting that soybean in these two countries form different gene pools. When comparing the Japanese soybean with that from seven different Chinese soybean ecotypes, 164–200 alleles were specific to the Chinese accessions and 64–112 specific to the Japanese accessions. The comparison of SSRs diversity revealed that accessions from China exhibited more genetic diversity than those from Japan. The data were analyzed to resolve the genetic structure and to interpret the evolutionary relationships between groups. Three distinct groups were identified, corresponding to Japanese soybean, Northern China soybean, Southern China soybean and a mixed group in which most accessions were from central China. The results indicate that accessions from Japan are distinct from Chinese ones, and Japanese accessions had more close relationship with Chinese northeast spring and southern spring ecotypes. We further analyzed five agronomic trait-related SSR loci and found that the preponderant alleles are different in Chinese and Japanese soybean. Our study provides important insights into further utilization of Japanese soybean in Chinese soybean breeding.     

Genetic diversity of melon landraces (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) in the Xinjiang Uygur Autonomous Region on the basis of simple sequence repeat markers

We investigated the genetic variation and relationships among 35 melon landraces collected from the Xinjiang Uygur Autonomous Region in northwestern China by using 19 polymorphic simple sequence repeat markers (SSRs). A total of 55 polymorphic alleles were amplified. The number of alleles per SSR locus ranged from 2 to 5 with an average of 2.89 alleles per locus. The average gene diversity (GD) was 0.42 with a range of 0.06–0.71, and the average observed heterozygosity was 0.22 with a range of 0.06–0.97, indicating that the genetic diversity among the Xinjiang melon landraces was abundant. Genetic variation was also detected between the landrace populations in different regions in Xinjiang. The most abundant genetic diversity was observed among the landraces in Eastern Xinjiang, with the highest GD of 0.45 and PIC of 0.39. Eleven alleles (20 %) were found exclusively in the landraces from Eastern Xinjiang, and two alleles (3.6 %) were unique to the landraces from Southern and Northern Xinjiang. The genetic similarity matrix was defined on the basis of Jaccard coefficient to determine the genetic relationships among Xinjiang landraces. Cluster analysis was performed using the unweighted pair group method with arithmetic means, showing that the ‘wild Hami’ (XJ-34) landrace was distinct from the 34 other landraces that were divided into three clusters. Therefore, the genetic background of XJ-34 differed from that of the other landraces. The landraces were not precisely separated on the basis of their geographic origins, although most of these landraces were likely grouped near one another, as visualized through principal coordinate analysis. Thus, western China is one of the primary or secondary centers of melon diversity because of the relatively higher genetic variation detected among Xinjiang landraces. Except the ‘wild Hami’ landrace, Xinjiang melon landraces could be classified into two botanical varieties, namely, var. inodorus and var. cantalupensis. However, the distinction between these two genotypes was not significantly different.     

利用SRAP和ISSR标记分析广东茶树种质资源的遗传多样性

利用EST-SSR标记对云南134份茶树资源遗传多样性和亲缘关系进行了分析。30对引物共检测到等位位点127个,平均每对引物产生423个;共检测到263个基因型,平均每对引物所扩增的基因型有88个;遗传多态性信息含量平均达0501,高于其它地区的相关研究结果,表明云南茶树资源具有丰富的遗传多样性。资源间的平均遗传距离和相似系数分别为0413和0597,说明资源间的遗传差异比较大,遗传基础较宽。聚类可将134份资源划分为4大组。8个种群间的遗传相似系数变异范围为0753～0981,平均遗传相似系数为0891,表明不同种群间的遗传差异比较小。云南茶树资源间亲缘关系的揭示为今后茶树资源保存和新品种选育提供了一定理论依据。     

EST-SSR分析云南茶树资源的遗传多样性和亲缘关系

利用EST-SSR标记对云南134份茶树资源遗传多样性和亲缘关系进行了分析。30对引物共检测到等位位点127个,平均每对引物产生423个;共检测到263个基因型,平均每对引物所扩增的基因型有88个;遗传多态性信息含量平均达0501,高于其它地区的相关研究结果,表明云南茶树资源具有丰富的遗传多样性。资源间的平均遗传距离和相似系数分别为0413和0597,说明资源间的遗传差异比较大,遗传基础较宽。聚类可将134份资源划分为4大组。8个种群间的遗传相似系数变异范围为0753～0981,平均遗传相似系数为0891,表明不同种群间的遗传差异比较小。云南茶树资源间亲缘关系的揭示为今后茶树资源保存和新品种选育提供了一定理论依据。     

EST-SSR based estimates on functional genetic variation in a barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) collection from Egypt

Genetic diversity was investigated in a set of 27 barley genotypes, originated from Egypt, using 23 expressed sequence tag (EST)-derived simple sequence repeats (eSSRs), representatives of the seven barley chromosomes. Ninety-five alleles were detected among all the barley accessions. For 22 polymorphic eSSRs, the number of alleles per locus varied from 2 to 12, with a mean of 4.318 alleles per locus. The lowest and the largest number of alleles per locus among the seven homeologous groups was observed in homeologous group 5H and 4H with 3.00 and 6.33, respectively. The gene diversity increased as the number of alleles increased. Gene diversity for 22 ESTs loci varied from 0.137 for GBM1404 to 0.896 for GBM1015 with an average of 0.563. A significant correlation coefficient between gene diversity and the number of alleles was high, r = 0.741 (P < 0.01). Cluster analysis was conducted based on eSSRs data to group the barley genotypes and to construct a dendrogram. Four groups can be distinguished by truncating the dendrogram at gs value of 0.77.     

Diversity in Phenotypic Profiles in Landrace Populations of Vietnamese Rice: A Case Study of Agronomic Characters for Conserving Crop Genetic Diversity on Farm

Phenotypic variation of agronomic characters in aromatic rice (Oryza sativa L.) was analyzed in order to elucidate the genetic diversity of the populations of rice landraces. Thirteen populations in Namdinh province, in the Red River Delta of Vietnam, were studied in both of farmers fields and an experimental field. Only small differences were found in agronomic characters among the populations, including populations containing different varieties. Variations among the populations in the frequency distributions of several characters suggested different degrees of diversity among the populations even among populations with the same variety name. The phenotypic profiles of farmers fields were different from those in an experimental field suggesting the environmental differences between them. One variety cultivated in the Myloc district had different phenotypic characters from the other varieties in the Haihau district more clearly in the experimental field. Since the small phenotypic variation in each of farm might be due to the genetic drift and selection by the farmers, on farm conservation of the landraces of rice is considered to be under a force to decrease phenotypic diversity. The present study suggests that the targeting several farms with different phenotypic profiles contribute to the conservation of regional genetic diversity of the landraces of rice.     

Evaluation of Genetic Diversity Among Bulgarian Winter Wheat (Triticum aestivum L.) Varieties During the Period 1925–2003 Using Microsatellites

Simple sequence repeats (SSRs) were used to study the genetic diversity within old and modern Bulgarian winter wheat (Triticum aestivum L.) varieties released in 20th century. A set of 91 varieties were screened by 19 wheat microsatellite markers (WMS), covering 17 wheat chromosomes, and one secalin-specific marker for rye chromosome arm 1RS. A total of 136 allelic variants were detected at 22 loci, ranging from 2 to 11, with an average of 6.8 alleles per marker. For 7 markers, null alleles were detected. The occurrence of rare alleles (frequency <2%) was observed for 13 markers. The polymorphism information content (PIC) values of the markers ranged from 0.10 (WMS0165 on 4AS) to 0.81 (WMS0437 on 7DL) with an average of 0.51. Approximately 74% of the varieties, mostly non-commercial, showed heterogeneity, with an average level of 10.1%. For the majority of markers, the relative frequencies of alleles varied considerably among different groups of varieties, revealing the effects of different selection between breeding centres. Some alleles, present in old genotypes, were lost, and new alleles have been introduced into modern varieties. Genetic diversity values over different periods of release were high, starting at 0.64 for varieties developed before 1960 to reach 0.71 in 1990s, revealing no declining trends in the diversity due to breeding activity. The cluster analysis discriminated all varieties (except for two) and revealed distinct groups of old and modern varieties, released from the main breeding centres in Northern, Southern and Western Bulgaria.     

Assessment of genomic diversity among and within Iranian confectionery sunflower (Helianthus annuus L.) populations by using simple sequence repeat markers

Abstract

Genetic variation in 15 confectionery sunflower populations from different geographical regions of Iran was evaluated using microsatellites. Forty-three alleles were scored at 20 SSR loci. The average number of alleles per locus (na) and the effective allele number (ne) were 2.15 and 1.72, respectively. Observed heterozygosity (0.55) was higher than the expected (0.38) indicating lack of Hardy–Weinberg equilibrium. The genetic similarity matrix for individuals was constructed using Kosman and Leonard similarity coefficient, and Nei's Genetic Identity as well as Kosman's genetic distance was calculated among populations. Dendrograms were constructed by the unweighted pair-group method using arithmetic average (UPGMA) algorithm. The analysis of molecular variance revealed lower genetic variation among populations (14%), than within populations (86%). The average number of migrants (Nm) between populations based on AMOVA (Fst = 0.062) was 4.79. Results displayed high variation among individual plants, possibly because of the high allogamy. Genetic similarity between populations in this study could be considered as consequences of genetic equilibrium that has occurred over the long period of cultivation of confectionery sunflower in this region as well as seed exchange among provinces. The traditional assumption that selecting genotypes of different geographical origin will maximize the diversity available to a breeding project does not hold in confectionery sunflower.     

微卫星标记分析我国南方常规水稻品种的遗传差异

利用12个DNA微卫星标记对2005-2007年参加南方稻区国家水稻品种区域试验的23个长江中下游早籼、晚粳和华南早籼常规品种进行标记检测,结果共检测到等位基因47个,平均每个标记3.9个,变幅2～7个,多态性频率（FP）平均值0.632,变幅0.403～0.842。遗传差异分析表明：长江中下游早籼与长江中下游晚粳(平均遗传相似系数AGSC=0.036)、华南早籼与长江中下游晚粳（AGSC=0.125）品种间的遗传差异明显,长江中下游早籼与华南早籼（AGSC=0.449）品种间的遗传差异中等,华南早籼（AGSC=0.606）、长江中下游早籼（AGSC=0.682）、长江中下游晚粳（AGSC=0.761）品种间的遗传差异较小。