甘蓝型油菜桔红花显性基因候选区域的NGS定位及InDel标记开发  

Location and InDel markers for candidate interval of the orange petal gene in Brassica napus L.by next generation sequencing

作　　者：郭青青 周蓉 陈雪 陈蕾 李加纳[1,2] 王瑞 GUO Qing-Qing;ZHOU Rong;CHEN Xue;CHEN Lei;LI Jia-Na;WANG Rui(College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China;Chongqing Engineering Research Center for Rapeseed,Chongqing 400715,China)

机构地区：[1]西南大学农学与生物科技学院,重庆400715 [2]重庆市油菜工程技术研究中心,重庆400715

出　　处：《作物学报》2021年第11期2163-2172,共10页Acta Agronomica Sinica

基　　金：西南大学校创项目(X202010635478);国家重点研发计划项目“七大作物育种”(2016YFD0101300)资助。

摘　　要：甘蓝型油菜花色的选育和改良已成为种质资源鉴定和材料创制的重要研究方向。目前为止,甘蓝型油菜桔红花显性基因定位的研究还未见报道。本研究以甘蓝型油菜双单倍体(doubled haploid,DH)纯系黄花Y05和桔红花R08杂交,分析桔红花性状遗传模式。在F_(2)群体中选取30株极端桔红花和30株极端纯黄花构建叶片DNA子代池和花瓣RNA子代池,对亲本和DNA子代池进行30×重测序,对RNA子代池进行6G测序。以法国甘蓝型油菜Darmor-bzh为参考序列,QTL-seq流程和MMAPPR流程相互结合鉴定桔红花基因候选区间。利用IGV软件可视化分析候选区间内插入缺失(InDel)变异位点,根据候选区间信息设计InDel引物。结果表明,桔红花性状受1对显性主效基因控制,全基因组重测序定位桔红花性状基因候选区间结果与转录组测序定位结果高度一致,均位于法国甘蓝型油菜Darmor-bzh A07染色体18~19 Mb。聚丙烯酰胺凝胶电泳筛选到3个与桔红花性状紧密连锁共分离的InDel标记。这些研究为精细定位桔红花显性候选基因以及分子标记辅助选育甘蓝型油菜桔红花新材料提供新思路。The petal color has been one of the major goals of breeding and genetic research in Brassica napus L.To date,there have been no reports about interval location of dominant orange petal gene trait in B.napus L.In this study,we con-structed an F_(2) mapping population with 458 individuals from the cross between DH Y05(yellow petal)and DH R08(orange petal).Whole-genome re-sequencing of DNAs and transcriptome sequencing of RNAs were from two populations each composed of 30 individuals showing extreme opposite trait for a given phenotype in a segregating progeny.Then we per-formed 30×and 6G of sequencing.Darmor-bzh as the reference genome was aligned to sequence data from the two bulks and parents.QTL-seq and Mutation Mapping Analysis Pipeline for Pooled RNA-seq(MMAPPR)workflow were applied to identify the candidate region of the orange petal gene.The insertion-deletion(InDel)sites can be visualized in candidate interval by Integrative Genomics Viewer(IGV).Based on these Indel variations,we used Vector and Blast to design InDel primers.The results indicated that the orange petal trait was controlled by a dominant major gene.A major candidate region was identified on chromosome A07(18-19 Mb)of Darmor-bzh.Three InDel markers linked to the orange petal gene were screened by Polyacrylamide gel electrophoresis(PAGE).This study may provide a novel idea for fine mapping dominant orange petal gene as well as marker assisted selection.

关 键 词：甘蓝型油菜 桔红花性状 基因定位 测序 分子标记 

分 类 号：S565.4[农业科学—作物学]