绿豆窄叶突变体vrnl9基因的精细定位与转录组分析  

Fine mapping and transcriptome analysis of a narrow leaf mutant gene vrnl9 in mungbean

作　　者：叶卫军 吴泽江 田东丰[1] 张阴 张玲玲 周斌[1] YE Wei-jun;WU Ze-jiang;TIAN Dong-feng;ZHANG Yin;ZHANG Ling-ling;ZHOU Bin(Crop Research Institute,Anhui Academy of Agricultural Sciences,Hefei 230031,China)

机构地区：[1]安徽省农业科学院作物研究所,安徽合肥230031

出　　处：《江苏农业学报》2024年第2期203-212,共10页Jiangsu Journal of Agricultural Sciences

基　　金：国家食用豆产业技术体系项目(CARS-08-Z11)。

摘　　要：叶片是绿豆最重要的光合作用场所,其形态结构影响光合作用、群体结构和产量。筛选和鉴定绿豆叶形突变体材料,为探究叶片发育的分子调控机理和叶形遗传改良奠定基础。在皖科绿3号EMS(Ethyl methyl sulphonate)诱变突变体库中鉴定出1个窄叶突变体vrnl9,并对该突变体进行表型鉴定、基因定位和转录组分析。表型分析发现,窄叶突变体vrnl9叶片宽较野生型皖科绿3号显著减小,叶面积和叶柄长也显著缩减;突变体主茎退化,生育期延长10 d。在突变位点的定位中,本研究利用苏绿16-10与突变体vrnl9杂交构建的F2群体进行遗传分析和基因精细定位。结果表明,窄叶突变体vrnl9的窄叶表型受单个隐性核基因控制(χ^(2)=1.40)。BSA测序分析将突变位点定位在第9染色体上0~3.1 Mb的区间内,结合图位克隆的方法,本研究将窄叶基因vrnl9定位在标记NL-15和NL-28之间354.6 kb的区间内。转录组学分析发现,与野生型皖科绿3号相比,突变体vrnl9中有182个基因的表达量发生显著改变,其中86个上调、96个下调。KEGG分析结果表明,差异表达基因显著富集在植物激素信号传导、萜类骨架的生物合成、玉米素的生物合成等代谢通路。这些研究结果为克隆窄叶基因vrnl9和解析绿豆叶片生长发育的分子调控机理提供理论依据。Leaf is the most important place for mungbean photosynthesis,and its morphological structure affects photosynthesis,population structure and yield in mungbean.Screening and identification of leaf shape mutants can lay the foundation for studying molecular regulation mechanism of leaf development and genetic improvement of leaf shape in mungbean.A narrow leaf mutant vrnl9 was identified from an ethyl methyl sulfonate(EMS)induced Wankelü3(WK3)mutant library.Phenotype identification,gene mapping and transcriptome analysis for vrnl9 were completed.Phenotype analysis showed that the leaf width,leaf area and petiole length of vrnl9 were significantly decreased compared with wild-type WK3.The main stem of mutant degenerated and the growth period was extended by ten days.In order to map the mutation site,the F 2 population constructed by the cross between Sulü16-10 and the mutant vrnl9 was used for genetic analysis and fine mapping.The narrow leaf trait of vrnl9 was controlled by a single recessive nuclear gene(χ^(2)=1.40).The BSA sequencing analysis showed that the mutation site was located within an interval of 0-3.1 Mb on chromosome 9.The narrow-leaf gene vrnl9 was finally narrowed down to a 354.6 kb region between the markers NL-15 and NL-28 by using map-based cloning strategy.Transcriptome analysis revealed that the expression levels of 182 genes in mutant vrnl9 were significantly changed compared with wild-type WK3,of which 86 genes were up-regulated and 96 genes were down-regulated.KEGG analysis results showed that the differentially expressed genes were significantly enriched in plant hormone signal transduction,terpenoid backbone biosynthesis,zeatin biosynthesis and other metabolic pathways.These results provide a theoretical basis for cloning vrnl9 and understanding the molecular regulation mechanism of leaf development in mung bean.

关 键 词：绿豆 窄叶 vrnl9基因 基因定位 转录组分析 

分 类 号：S522[农业科学—作物学]