基于10个棉花腺体相关材料转录组的EST-SSR标记开发  

Development of EST-SSR Markers in Cotton Based on the Transcriptome Information of 10 Gland-related Materials

作　　者：程海亮[1] 陆才瑞[1] 邹长松[1] 余道乾[1] 姜鹏飞[1] 杨文翠[1] 张友平[1] 王巧连 宋国立[1] 

机构地区：[1]中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000

出　　处：《棉花学报》2015年第1期1-8,共8页Cotton Science

基　　金：国家自然科学基金(31271768;31301369)

摘　　要：棉花是重要的经济作物,但有毒棉酚的存在,使棉子得不到充分的利用,所以低酚棉育种是棉花育种的重要内容之一。色素腺体是棉酚的储存器官,开发功能型分子标记,加密棉花遗传图谱对色素腺体和其他重要性状相关基因的研究具有重要作用。本试验以10个有腺体和无腺体材料转录组信息为基础开发EST-SSR标记,在12895条1 kb以上的Unigene中共搜索到1546个SSR位点,发生频率和平均距离分别是11.99%和15.99 kb。得到的EST-SSR中,二碱基和三碱基重复是主要的重复类型,分别占30.85%和48.97%,AT/TA和GAA/CTT分别是二碱基和三碱基的优势重复类型。对长度≥20 bp的SSR共设计合成了56对引物,在这10个材料中进行检测,能扩增出清晰条带的有38对,占67.86%,其中呈多态性的有9对,占23.68%,并对其所在的Unigene功能和表达量进行了初步分析。本研究进一步证明了棉花EST-SSR标记开发的可行性,并且为棉花高密度遗传图谱和腺体相关基因的研究奠定了基础。Cotton is an important economic crop, however, the cottonseed is underutilized because of the presence of toxic gossy- pol, so low gossypol cotton breeding is an important part of the cotton breeding. The development of molecular markers and construction of high resolution genetic map will play an important role in research of genes relative to pigment glands, which are storage organ of gossypol, and other important traits. In this study, we developed EST-SSR markers based on the transcriptome information from 10 gland and glandless materials. 1546 EST-SSRs were obtained from 12895 unigenes more than 1 kb in length, which accounts for 11.99%, and the average distance of SSRs was 15.99 kb. Among the obtained EST-SSRs, dinu- cleotide and trinucleotide repeats were the dominant types, accounting for 30.85% and 48.97% respectively. AT/TA and GAA/CTT were the most abundant motifs respectively in dinucleotide and trinucleotide repeats. Total of 56 pairs of primers were designed and synthesized for the SSRs more than 20 bp and tested in the 10 materials to evaluate the application and poly- morphism. 38 pairs of primers could produce clear bands in PAGE electrophoresis, accounting for 67.86%, and 9 EST-SSRs a- mong them showed polymorphism, accounting for 23.68%. Further more, the functions and expression of the unigenes contain- ing polymorphic EST-SSR were analyzed preliminarily. This study further proves that it is feasible to develop EST-SSR markers in cotton, and also basic work for the research of cotton high resolution genetic map and pigment gland related genes.

关 键 词：棉花 EST.SSR标记 多态性 色素腺体 

分 类 号：S562[农业科学—作物学]