基于叶绿体InDel标记对玉米S型胞质不育制种鉴定的研究

Identification of Maize S-type Cytoplasmic Male Sterile(CMS) Using Two Chloroplast InDel Markers in Seed Production

以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个和5个碱基的插入缺失变异,属于trnS-trnG和ndhJ-ndhK基因间区。CPMIDP01CE和CP?MIDP02CE引物的扩增产物如果含有83 bp插入序列(片段长度为339 bp),不含有5 bp的插入序列(片段长度为239 bp),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。

Using 29 representative maize inbred lines with rich phenotypical and genotypical diversity together with triplets materials,the primers of 11 In Del loci from chloroplast genome were designed,screened and evaluated,and two specific pairs of primers(CPMIDP01 CE and CPMIDP02 CE)for identifying maize S-type cytoplasmic male sterile(CMS)were obtained.The variations identified by CPMIDP01 CE and CPMIDP02 CE result from the insertion/deletion of 83 and 5 base pairs(bp)locating on the intergenic region of trn S-trn G and ndh J-ndh K.If the amplified products of CPMIDP01 CE and CPMIDP02 CE primers were 339 and 239 bp respectively,meaning the amplified ma-terials contain a 83-bp insertion sequence,but lack of a 5-bp insertion sequence,the analysis samples are S-typeCMS 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 Kompetitive allele specific PCR(KASP),Taq Man 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 established by two chloroplast In Del markers,and it will provide key technical support for the production of maize hybrids in China.