基于BSA-seq技术对番茄封顶花序节位基因的精细定位  

Fine Mapping of Inflorescence Pruning Node Gene in Tomato(Solanum lycopersicum)Based on BSA-seq Technique

作　　者：师海林 张丹丹 高倩[4] 尤园园 舒金帅[5] 王帅[1,2,3] 毛秀杰 SHI Hai-Lin;ZHANG Dan-Dan;GAO Qian;YOU Yuan-Yuan;SHU Jin-Shuai;WANG Shuai;MAO Xiu-Jie(College of Horticultural Science and Technology,Hebei Normal University of Science and Technology,Qinhuangdao 066004,China;Hebei Key Laboratory of Characteristic Horticultural Germplasm Mining and Innovative Utilization,Qinhuangdao 066004,China;Hebei Higher Institute Application Technology Research and Development Center of Horticultural Plant Biological Breeding,Qinhuangdao 066004,China;Changli Institute of Pomology,Hebei Academy of Agricultural and Forestry Sciences,Changli 066000,China;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]河北科技师范学院园艺科技学院,秦皇岛066004 [2]河北省特色园艺种质挖掘与创新利用重点实验室,秦皇岛066004 [3]河北省高校特色园艺植物生物育种应用技术研发中心,秦皇岛066004 [4]河北省农林科学院昌黎果树研究所,昌黎066000 [5]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《农业生物技术学报》2023年第7期1357-1367,共11页Journal of Agricultural Biotechnology

基　　金：河北省重点研发计划项目(20326325D);河北省省属高校基本科研业务费专项(2021JK07)。

摘　　要：主茎花序数的多少影响着自封顶类型番茄(Solanum lycopersicum)的产量,精细定位其封顶花序节位基因,可为进一步基因克隆以及功能分析、调控机理解析提供依据。本研究以低节位自封顶番茄品系AXF和高节位自封顶番茄品系GXF为亲本构建F2分离群体,应用混合群组分离分析测序(bulked segregant analysis sequencing,BSA-seq)和分子标记方法进行了连锁分析及基因定位。结果表明,关联区间位于2号染色体47.56~47.59 Mb,大小约为25497 bp,含有3个候选基因(硫代硫酸硫转移酶18(thiosulfatesul furtransferase 18,TST18)(GenBank No.XM_004232452.4),MLO类家族蛋白4(MLO-like protein 4,MLO4)(GenBank No.XM_019212390.2)和26S蛋白酶体非ATP酶调节亚基4(26S proteasome non-ATPase regulatory subunit 4,PSMD4)(GenBank No.XM_010318126.3))。进一步开发118对InDel(insertion-deletion)/CAPS(cleaved amplified polymorphism sequences)/dCAPS(derived CAPS)引物进行精细定位,筛选获得1对多态性高的dCAPS14分子标记,该标记与PSMD4紧密连锁。qRT-PCR分析表明该候选基因在两个亲本不同组织中的表达量存在显著性差异。PSMD4功能为参与泛素-蛋白酶体系统介导的蛋白降解,可能对番茄封顶花序节位起着重要调控作用。本研究对分子辅助选育易于栽培操作的理想型自封顶番茄品种具有重要意义。The number of main stem flowers affects the yield of self-pruning tomato(Solanum lycopersicum).In order to locate the genes associated with inflorescence pruning node,a fine mapping was carried out for laying a foundation for gene cloning and functional analysis and providing evidences for the analysis of regulation mechanism.In this study,one F2 population was constructed from the cross between AXF(low nodal self-pruning tomato strain)and GXF(nodal self-pruning tomato strain),and linkage analysis and gene localization were performed by bulked segregant analysis sequencing(BSA-seq)and molecular marker.The results showed that the associated region from 47.56~47.59 Mb was located on chromosome 2 with the size of 25497 bp,containing 3 candidate genes(thiosulfate sulfurtransferase 18(TST18)(GenBank No.XM_004232452.4),MLO-like protein 4(MLO4)(GenBank No.XM_019212390.2)and 26S proteasome non-ATPase regulatory subunit 4(PSMD4)(GenBank No.XM_010318126.3)).118 InDel(insertion-deletion)/CAPS(cleaved amplified polymorphism sequences)/dCAPS(derived CAPS)primers were designed for fine mapping and a dCAPS14 molecular maker closely linked with candidate gene PSMD4 was finally delimited by further analysis.qRT-PCR analysis showed that the relative expression of the candidate gene was significantly different between AXF and GXF.The PSMD4 was involved in ubiquitin-proteasome system mediated protein degradation and served as key candidate gene.PSMD4 might play an important role in the controlling of inflorescence pruning node.These results had a significance in marker-assisted self-pruning tomato breeding for ideal architecture with easy cultivation.

关 键 词：自封顶番茄 混合群组分离分析测序(BSA-seq) 分子标记 基因定位 

分 类 号：S641.2[农业科学—蔬菜学]