莴笋红叶基因图位克隆及分子标记开发  

Map-Based Cloning and Development of Molecular Markers of Red-Leaf Gene in Stem Lettuce(Lactuca sativa var.angustata)

作　　者：李春 何振 刘小俊[1,2] 梁根云 李艺凡[1,2] 杨楠 蔡鹏 李跃建 房超[1,2] 刘独臣 LI Chun;HE Zhen;LIU Xiaojun;LIANG Genyun;LI Yifan;YANG Nan;CAI Peng;LI Yuejian;FANG Chao;LIU Duchen(Vegetable Germplasm Innovation and Variety Improvement Key Laboratory of Sichuan Province,Horticulture Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,China;Pengzhou Rural Investment Development Co.,Ltd.,Chengdu 611934,China;Rice Research Institute,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川省农业科学院园艺研究所蔬菜种质与品种创新四川省重点实验室,成都610066 [2]彭州市乡村投资发展有限公司,成都611934 [3]四川农业大学水稻研究所,成都611130

出　　处：《四川农业大学学报》2024年第3期523-528,共6页Journal of Sichuan Agricultural University

基　　金：四川省蔬菜育种攻关项目(2021YFYZ0022);现代农业产业技术体系专项资金(CARS-23-G38);国家现代农业产业技术体系四川蔬菜创新团队(川农函[2019]427号)。

摘　　要：【目的】研究莴笋叶色变异的遗传基础,加速莴笋新品种育种进程,提高育种效率。【方法】通过构建红叶和绿叶莴笋杂交群体,分析遗传规律,利用混池分组分析法(BSA-seq)和图位克隆法精细定位并克隆了莴笋红叶基因Lactuca sativa Red Leaf 1(LsRL1),根据变异位点设计了分子标记用于红叶莴笋分子标记辅助育种。【结果】遗传分析结果显示F2分离群体中红叶个体与绿叶个体的分离比为3∶1,说明研究中莴笋叶色表型受一个基因控制,且红叶相对于绿叶为显性性状。混池分组分析法(BSA-seq)将莴笋叶色基因LsRL1初步定位在莴笋第5染色体336.00 Mb到339.64 Mb的范围内。利用图位克隆的方法进一步将LsRL1基因的范围缩小至2个Indel分子标记ls06和ls12之间的85.17 kb区间内,这一区间内包含2个基因LOC111892298和LOC111892911。亲本间差异位点分析将LsRL1的候选基因确定为LOC111892911,该基因在拟南芥中的同源基因编码一个bHLH转录因子,参与花青素合成途径的调控。【结论】基于LsRL1基因在2个亲本间的插入缺失变异,我们开发了一个可用于红叶莴笋分子标记辅助育种的Indel标记,以加速红叶莴笋的育种进程。【Objective】In order to study the genetic basis of leaf color of stem lettuce,accelerate the breeding process of red-leaf stem lettuce varieties and improve the breeding efficiency.【Method】In this study,a hybrid population of red-leaf and green-leaf stem lettuce was constructed,the genetic rule was analyzed,and the bulk segregant analysis(BSA-seq)and map-based cloning were used to finely map the red leaf gene Lactuca sativa Red Leaf 1(LsRL1),and a molecular marker was designed for marker-assisted breeding of red-leaf stem lettuce.【Result】Genetic analysis showed that the segrega⁃tion ratio of red-leaf individuals to green-leaf individuals in the F2 segregating population was 3∶1,indi⁃cating that the leaf color in this study was controlled by a single gene,and red leaf was a dominant trait relative to green leaf.The bulk segregant analysis(BSA-seq)method preliminarily mapped the leaf color gene LsRL1 to the range of 336.00 Mb to 339.64 Mb on chromosome 5 of stem lettuce. Using the map based cloning method, the range of LsRL1 gene was further narrowed down to an 85.17 kb interval be⁃ tween two Indel molecular markers ls06 and ls12, which contained two genes LOC111892298 and LOC111892911. Sequence difference analysis identified LOC111892911 as the candidate gene for LsRL1, which encodes a bHLH transcription factor in Arabidopsis thaliana and is involved in the regula⁃ tion of anthocyanin synthesis pathway. 【Conclusion】 Based on the insertion/deletion variation of LsRL1 gene between two parents, we developed an Indel marker that can be used for molecular marker-assisted breeding of red-leaf stem lettuce to accelerate the breeding process.

关 键 词：莴笋 红叶 图位克隆 分子标记 

分 类 号：S636.2[农业科学—蔬菜学]