InDel分子标记WC656的开发及其在鉴别小麦-簇毛麦5VS纯合易位中的应用  

Development of Insertion-Deletion Molecular Marker WC656 and Its Application in Identifying Wheat-Dasypyrum villosa 5VS Homozygous Translocation

作　　者：郭江涛 吕远大[2] 赵姝楠 周淼平[1] 姚金保[1] 杨学明[1] GUO Jiangtao;L Yuanda;ZHAO Shunan;ZHOU Miaoping;YAO Jinbao;YANG Xueming(Institute of Food Crops/Jiangsu Provincial Key Laboratory of Agrobiology,Jiangsu Academy of Agricultural Sciences,Nanjing,Jiangsu 210014,China;Excellence and Innovation Center,Jiangsu Academy of Agricultural Sciences,Nanjing,Jiangsu 210014,China;State Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing Agricultural University,Nanjing,Jiangsu 210095,China)

机构地区：[1]江苏省农业科学院粮食作物研究所/江苏省农业生物学重点实验室,江苏南京210014 [2]江苏省农业科学院卓越创新中心,江苏南京210014 [3]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《麦类作物学报》2023年第9期1115-1124,共10页Journal of Triticeae Crops

基　　金：江苏省农业科技自主创新资金项目[CX(19)1001];江苏省重点研发计划(现代农业)项目(BE2018350)。

摘　　要：簇毛麦是小麦品种改良重要的遗传资源,其5VS上含有硬度基因Dina/Dinb、抗白粉病基因Pm55和抗条锈病基因Yr5V,创制普通小麦-簇毛麦5VS易位系新种质,对于高效利用5VS上的有益基因进行小麦品种遗传改良具有重要意义。为了便于鉴别普通小麦-簇毛麦5VS纯合易位,本研究以mInDel软件分析普通小麦中国春基因组序列,开发了小麦第5同源群短臂的InDel特异分子标记WC656,对小麦品种以及普通小麦-簇毛麦5VS.5AL和5VS.5DL易位系及其衍生后代进行PCR扩增,根据扩增片段大小来区分5AS、5BS、5DS染色体臂以及5VS易位系。结果表明,在21个普通小麦品种、小麦-簇毛麦5VS回交F 2代中杂合易位单株,均扩增出379 bp(5AS)、471 bp(5BS)、422 bp(5DS)3条带。在T5VS.5AL高代品系、回交F 2代中纯合易位单株扩增出471 bp(5BS)、422 bp(5DS)2条带,379 bp(5AS)条带缺失。T5VS.5DL高代品系、回交F 2代中纯合易位单株扩增出379 bp(5AS)、471 bp(5BS)2条带,422 bp(5DS)条带缺失。WC656鉴别的5VS纯合易位结果与顺序FISH-GISH分析结果一致。T5VS.5AL、T5VS.5DL具有抗白粉病、籽粒硬度指数低等优良特性。因此,利用InDel分子标记WC656可以鉴别普通小麦-簇毛麦T5VS.5AL、T5VS.5DL衍生后代中的纯合易位,PCR扩增实验操作相对简便,可以为分子标记辅助选育携有5VS纯合易位的软质弱筋小麦提供帮助。Dasypyrum villosum(2n=14,VV)is one of important genetic resources for wheat cultivar improvement.Previous studies indicated that D.villosum chromosome arm 5VS confers genes including grain hardness gene Dina/Dinb,powdery mildew resistance gene Pm55 and yellow rust resistance gene Yr5V.It is of great significance to create Triticum aestivum-D.villosum 5VS homozygous translocation for wheat improvement utilizing these desired genes.In order to identify wheat-D.villosum 5VS homozygous translocation,a total of 265 InDel molecular markers were designed for detecting chromosome short arms for wheat homologous chromosomes 5 by mInDel software based on Chinese Spring reference genome.In this study,marker WC656 was finally used to discriminate 5AS,5BS,and 5DS of wheat chromosome according to the fragment size 379 bp,471 bp and 422 bp of PCR amplification detected on agarose gel electrophoresis,respectively.Three bands specific to 5AS,5BS,and 5DS were detected in 21 common wheat cultivars and wheat-D.villosum 5VS heterozygous translocation of two BC 4 F 2 individuals.The bands specific to 5BS and 5DS were also detected in homozygous translocation T5VS.5AL of BC 4 F 2 individuals and advanced lines,and the band of 5AS was absent.Similarly,the bands specific to 5AS and 5BS were detected in homozygous translocation T5VS.5DL of BC 4 F 2 individuals and advanced lines,and the band of 5DS was absent.The sequential FISH and GISH analyses further confirmed the results based on molecular marker WC656.Consequently,wheat-D.villosum 5VS homozygous translocation can be identified efficiently by marker WC656 in this study.T5VS.5AL and T5VS.5DL showed higher resistance to powdery mildew,and significantly lower grain hardness index compared to their recurrent parents.The PCR products can be detected by agarose gel electrophoresis,and the experimental process is relatively efficient.It will be helpful to expedite marker-assisted selection in soft and weak gluten wheat breeding program with wheat-D.villosum 5VS.5AL and 5VS.5DL translocation

关 键 词：小麦 簇毛麦 5VS纯合易位 InDel分子标记 

分 类 号：S512.1[农业科学—作物学] S330