黄瓜线粒体基因组SSR标记开发及其在种子纯度鉴定中的应用  

Development of mt SSR markers in cucumber(Cucumis sativus L.)and their application in the identification of seed purity

作　　者：李海梅[1] 沈佳[1] 赵娟[1] 娄群峰[1] 李季[1] 陈劲枫[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室/园艺学院,江苏南京210095

出　　处：《南京农业大学学报》2015年第5期764-771,共8页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金重点项目(31430075);国家重点基础研究发展计划项目(2012CB113900);国家863计划项目(2012AA100202);国家公益性行业(农业)科研专项(201403032);"十二五"国家科技支撑计划项目(2013BAD01B04-10);江苏省农业科技自主创新基金项目(CX(11)1002)

摘　　要：[目的]基于黄瓜线粒体父系遗传特性,研究线粒体基因组SSR(mitochondria genome simple sequence repeat,mt SSR)标记技术在黄瓜杂交种纯度鉴定上的可行性。[方法]根据黄瓜线粒体基因组序列开发特异性标记,选用不同生态型的黄瓜品种对标记进行筛选,利用多态性引物对人为掺假的黄瓜F1种子进行纯度检测。[结果]从72对标记中,共筛选出4对在30份黄瓜品种呈现出多态性的引物,其中mt SSR4引物扩增出102和107 bp的多态性片段;mt SSR10引物扩增出196、224和242 bp的多态性片段;mt SSR29引物扩增出226、239和280 bp的多态性片段;mt SSR44引物扩增出184和194 bp的多态性片段。利用引物mt SSR4和mt SSR10对4份黄瓜F1种子纯度进行鉴定,结果显示黄瓜F1种子的带型与父本的一致,可将掺入杂交组合中的假种子区分开。田间试验数据表明:分子标记检测与田间鉴定结果基本一致,可用于黄瓜杂交种种子纯度的鉴定。[结论]线粒体基因组特异序列SSR标记能够准确地鉴定黄瓜杂种一代种子,在保证黄瓜种子生产质量方面具有广泛的应用前景。[Objectives]Based on the paternal genetic characteristics of cucumber mitochondria,the purpose of this paper is to study the feasibility of mitochondria genome simple sequence repeat（ mt SSR） in hybrid seed purity identification of cucumber. [Methods]The specific markers were developed according to the mitochondrial genome sequence of cucumber. Markers were screened with different ecotypes of cucumber varieties,and then the purity of F1 hybrid seeds was detected with polymorphic primers. [Results]Four pairs of polymorphic primers were screened from 72 pairs of mt SSR markers in 30 cucumber varieties. The PCR amplification of mt SSR4 primer could distinguish two polymorphic fragments of 102 and 107 bp. Three polymorphic fragments of 196,224 and 242 bp were obtained with mt SSR10 primer. Three polymorphic fragments of 226,239 and 281 bp could be amplified by mt SSR29 primer.Besides,mt SSR44 primer could amplify two polymorphic fragments of 184 and 194 bp. Cucumber purity test of four F1 hybrids showed that the bands of F1 hybrids were consistent with the male parent respectively and the results tested in field. Contamination of F1 hybrid seeds caused by self-inbred and other unknown pollens can be detected. [Conclusions]The purity of cucumber F1 hybrids was accurately identified by mitochondrial genome sequence-specific SSR markers. It is suggested that mitochondrial genome SSR technique has application prospects to ensure production quality of cucumber seeds.

关 键 词：黄瓜 线粒体基因组 SSR分子标记 种子 纯度鉴定 

分 类 号：S642.2[农业科学—蔬菜学]