基于BSA-Seq和RNA-Seq方法鉴定大豆抗豆卷叶螟候选基因  

Identification of the candidate genes of soybean resistance to bean pyralid(Lamprosema indicata Fabricius)by BSA-Seq and RNA-Seq

作　　者：曾维英[1] 赖振光 孙祖东[1] 杨守臻[1] 陈怀珠[1] 唐向民[1] ZENG Wei-Ying;LAI Zhen-Guang;SUN Zu-Dong;YANG Shou-Zhen;CHEN Huai-Zhu;TANG Xiang-Min(Institute of Economic Crops,Guangxi Academy of Agricultural Sciences/Southwest Experimental Station of Maize-Soybean Intercrop,Ministry of Agriculture and Rural Affairs,Nanning 530007,Guangxi,China)

机构地区：[1]广西农业科学院经济作物研究所/农业农村部西南玉米大豆间套作区农业科学观测实验站,广西南宁530007

出　　处：《作物学报》2021年第8期1460-1471,共12页Acta Agronomica Sinica

基　　金：广西自然科学基金项目(2017GXNSFDA198037);广西农业科学院科技发展基金项目(桂农科2020YM116,2015YT58)资助。

摘　　要：豆卷叶螟(Lamprosema indicata Fabricius)是重要的大豆食叶性害虫,挖掘大豆抗豆卷叶螟相关基因对大豆抗虫品种选育和遗传改良至关重要。本研究用大豆高抗豆卷叶螟材料赶泰-2-2和高感豆卷叶螟材料皖82-178进行杂交构建F2代分离群体,从303个F2代单株中挑选出高抗豆卷叶螟和高感豆卷叶螟的单株各30株,分别构建2个极端性状的DNA混合池用于全基因组重测序以分析控制豆卷叶螟相关的候选基因。结果表明,4个样本中共有11,963,077个单核苷酸多态性(SNPs)标记,根据SNP-index方法关联分析,共有329个基因位于99%置信区间外,这些基因主要集中在7号染色体5,601,065~5,865,237 bp区间(总长为0.26 Mb)、16号染色体2,975,110~6,336,096 bp区间(总长为3.36 Mb)、18号染色体44,366,115~54,297,600 bp区间(总长为9.93 Mb)等区域内。将BSA-Seq结果与转录组测序结果进行联合分析发现,有12个基因相关联;最后,结合生物信息学分析、候选基因的表达模式和基因的同源注释,锁定CNGC4、WRKY16转录因子、AAP7、丝氨酸/苏氨酸蛋白激酶、ZPR1B等12个基因为控制豆卷叶螟性状相关的候选基因。本研究结果不仅为解析大豆抗豆卷叶螟的分子机理奠定重要的基础,也将为大豆抗虫基因的克隆奠定坚实的理论基础。Bean pyralid is an important leaf-feeding insect in soybean.Identification of insect-tolerant genes from soybean has great significant to the crop insect-tolerant breeding and genetic improvement.In this study,an F2 population with 303 individuals was constructed using insect-resistant line Gantai-2-2 and insect-sensitive line Wan 82-178.30 F2 insect-resistant individuals and 30 insect-sensitive individuals were selected respectively to construct two DNA pools which were used for the whole-genome re-sequencing.The results showed that there were a total of 11,963,077 single nucleotide polymorphism(SNPs)markers identified in two parental lines and two mixed pools.According to the association analysis of SNP-index method,a total of 329 genes were located outside the 99%confidence interval.These genes were mainly concentrated in the regions of 5,601,065–5,865,237 bp with a total of 0.26 Mb on chromosome 7,2,975,110–6,336,096 bp with a total of 3.36 Mb on chromosome 16,and 44,366,115–54,297,600 bp with a total of 9.93 Mb on chromosome 18.Correlation analysis of BSA-Seq and transcriptome sequencing showed that 12 genes were correlated.Then,12 candidate genes,including CNGC4,WRKY transcription factor 16,AAP7,serine/threonine protein kinase and ZPR1B were identified by bioinformatics analysis,differential expression analysis,and homologous annotation.This study laid an important foundation for the analysis of the molecular mechanism of soybean resistance to bean pyralid and the cloning of anti-insect genes.

关 键 词：大豆 豆卷叶螟 BSA-Seq RNA-SEQ 候选基因 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]