基于叶绿体InDel标记对玉米S型胞质不育制种鉴定的研究
本研究精选表型和基因型丰富的29份代表性玉米自交系以及三联体验证材料,基于叶绿体基因组中11个InDel位点进行引物设计、筛选评估,确定2对玉米S型不育鉴定特异引物。两对引物CPMIDP01CE和CPMIDP02CE的多态分别为83和5个碱基的插入缺失变异,属于trnS-trnG和ndhJ-ndhK基因间区。CPMIDP01CE和CPMIDP02CE引物的扩增产物如果含有83bp插入序列(片段长度为339bp),不含有5bp的插入序列(片段长度为239bp),所分析样品即为S型胞质不育材料。两对特异鉴定引物基于变性荧光毛细管、非变性凝胶毛细管、琼脂糖等不同电泳平台均建立了检测方法；并且具有兼容更高效的KASP、TaqMan等技术体系的潜力。本研究基于叶绿体InDel标记建立玉米S型胞质不育制种鉴定的一种简便快捷、稳定高效、低成本、创新性方法,为规模化杂交种生产提供重要的技术保障。
英文摘要:
In this study, using 29 representative maize inbred lines with rich phenotypes and genotypes and triplets materials, the primers of 11 InDel loci from chloroplast genome were designed, screened and evaluated, and two specific pairs primers to identify the maize S- type CMS were obtained. The variations of the two primers CPMIDP01CE and CPMIDP02CE are the insertion/deletion of 83 and 5 base pairs (bp), and belong to the intergenic region of trnS-trnG and ndhJ-ndhK. The amplified products of CPMIDP01CE and CPMIDP02CE primers contain 83bp insertion sequence (fragment length is 339bp), do not contain 5bp insertion sequence ( fragment length is 239bp), the analysis samples are S-type CMS materials. The detection methods were established using the two specific primers based on different electrophoresis platforms, such as denaturing fluorescence capillary, non-denatured gel capillary, agarose electrophoresis platform. Moreover, the two primers have the potential to be compatible with more efficient KASP, TaqMan and other technical systems. In this research, in order to identify maize S-type CMS materials, a simple-fast, stable-efficient, low cost and innovative method was constructed by chloroplast InDel markers, and it will provide key technical support for the production of maize hybrids in China.
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