基于中华猕猴桃“红阳”转录组序列开发EST-SSR分子标记(英文)  

Development of EST-SSR markers in Actinidia chinesis cv ‘Hongyang' based on transcriptomic sequences

作　　者：孟蒙[1] 唐维[1,2] 刘嘉[2] 黄胜雄[2] 余进德[1] 刘方方[1] 林琳[1] 张霞[1] 刘永胜[1,2] 

机构地区：[1]四川大学生命科学学院/水力学与山区河流开发保护国家重点实验室/生物资源与生态环境教育部重点实验室,成都610064 [2]合肥工业大学生物与食品工程学院,合肥230009

出　　处：《应用与环境生物学报》2014年第4期564-570,共7页Chinese Journal of Applied and Environmental Biology

基　　金：Supported by the Key Project from the Government of Sichuan Province(2013NZ0014);the Key Project from the Government of Anhui Province(2012AKKG0739);the National Basic Research Program of China(973 Program)(2011CB100401)~~

摘　　要：为开发猕猴桃EST-SSR标记,了解28个品种猕猴桃间的遗传多样性和遗传关系,对中华猕猴桃"红阳"的转录组序列进行分析,并根据分析结果设计SSR引物.之后采用CTAB法提取28个品种猕猴桃的DNA作为SSR-PCR的扩增模板,并根据扩增结果进行聚类分析.研究中共得到包含SSR的序列21 848条,其中重复单元为单碱基、双碱基、三碱基、四碱基、五碱基和六碱基的序列分别为1 642、15 965、3 141、248、368和484条,随机选择其中46条序列设计SSR引物.根据初步的PCR扩增,筛选出32对条带较少且明亮的引物分别对28个品种猕猴桃的DNA样本进行扩增,并对引物对应的SSR序列进行定位.结果显示,32对引物对应的序列中有19条能够得到完整的所在基因、染色体以及染色体中具体位置的信息.这些引物中有26对具有多态性,共统计到等位基因120个,每对引物得到1-11个等位基因,平均3.75个.28个猕猴桃品种之间的遗传相似性系数在0.53-0.97之间,在遗传相似系数为0.72的水平上,可将它们分为5大类,分类结果与传统形态学的划分基本一致.本研究揭示的各样本间的遗传关系可为未来猕猴桃的种质改良提供依据.A transcriptome analysis of kiwifruit（Actinidia chinese） cv ‘Hongyang＇ was conducted to develop EST-SSR markers and analyze the genetic diversity and relationship among the 28 kiwifruit samples. Genomic DNA of 28 Actinidia samples were extracted with CTAB method and used as a template in SSR-PCR analysis and cluster analysis. In total, 21 848 SSR-containing sequences were obtained, of which 1 642, 15 965, 3 141, 248, 368 and 484 were mono-, di-, tri-, tetra-, penta- and hexa-nucleotide, respectively. Forty-six pairs of flanking SSR primers were developed based on the SSR sequences detected. Based on the results of the PCR analysis, 32 pairs of SSR primers were selected because they provided a relatively low number of clearly amplificated bands. We obtained the precise gene ID, chromosome No. and both start and end sites of 19 sequences. Out of the 32 SSRs, 26 were polymorphic and 120 alleles were observed. Each pair of primers amplified 1-11 alleles with an average of 3.75 alleles per SSR. Fifteen specific bands were detected in 11 samples. The dice similarity coefficient of 28 samples ranged from 0.53 to 0.97. Clustering analysis suggested that the 28 Actinidia samples could be divided into 5 groups with a dice similarity coefficient 0.72. The grouping result was in general agreement with the traditional morphology-based classification. The genetic relatedness identified in the analyzed Actinidia samples provides potential clues for the selection of germplasm of interest for crossbreeding and variety improvement.

关 键 词：中华猕猴桃 遗传分析 转录组 重复单元 SSR 种质资源 

分 类 号：S663.4[农业科学—果树学]