BrGOLDEN基因调控大白菜金黄色内叶的遗传和分子机制研究  

Research on the Genetic and Molecular Mechanisms of BrGOLDEN Gene in Regulating the Golden Inner Leaves of Chinese Cabbage

作　　者：宋超 章时藩 李菲[1] 张慧[1] 孙日飞[1] 刘伟[1,2] 李国亮 张淑江[1] SONG Chao;ZHANG Shifan;LI Fei;ZHANG Hui;SUN Rifei;LIU Wei;LI Guoliang;ZHANG Shujiang(Institute of Vegetable and Flower,Chinese Academy of Agricultural Sciences/National Key Laboratory of Vegetable Biological Breeding,Beijing 100081,China;China Vegetable Seed Industry Technology(Nanning)Co.,Ltd.,Nanning 530002,China)

机构地区：[1]中国农业科学院蔬菜花卉研究所/蔬菜生物育种全国重点实验室,北京100081 [2]中蔬种业科技(南宁)有限责任公司,广西南宁530002

出　　处：《蔬菜》2025年第1期108-121,共14页Vegetables

基　　金：中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS);农业农村部园艺作物生物学与种质创制重点实验室项目;国家大宗蔬菜产业技术体系项目(CARS-23-A-14)。

摘　　要：金黄心大白菜和橘红心大白菜都富含类胡萝卜素,颜色亮丽,广受欢迎,因此,内叶颜色成为大白菜新品种选育的重要目标性状之一,但是其调控机制尚不清晰。本研究利用金黄心和橘红心大白菜构建回交群体,并通过开发金黄心性状的BrGOLDEN基因和橘红心性状的Bror基因性状的分子标记,对BrGOLDEN和Bror的互作规律进行初步的分析;对成熟期金黄心大白菜和浅黄心大白菜的内叶进行转录组测序(RNA-seq)分析,了解类胡萝卜素代谢途径相关基因的表达;对BrGOLDEN启动子进行克隆并分析关键顺式作用元件,同时利用5'截短启动子克隆及GUS染色探究BrGOLDEN启动子的核心功能区域。遗传规律结合分子标记结果表明:显性基因BrGOLDEN与隐性基因Bror非独立遗传,Bror基因对BrGOLDEN基因表现为上位效应,即BrGOLDEN基因与Bror同时存在时,仅表现橘红心表型。转录组数据显示:金黄心大白菜中2-C-甲基-D-赤藓糖醇-4-磷酸(MEP)前体合成途径及类胡萝卜素合成途径相关基因的表达量基本都比浅黄心大白菜高;类胡萝卜素合成途径中合成八氢番茄红素聚合酶(PSY)、玉米黄质环氧化酶(ZEP)的基因表达量基本都高于途径上其他的基因,表明BrGOLDEN基因可能通过调控类胡萝卜素合成途径的起始和结束的相关基因来控制金黄心大白菜合成更多的类胡萝卜素。启动子功能分析结果表明:在启动强度上,不同缺失长度的启动子表现不同,并且BrGOLDEN基因启动子的核心作用元件可能在-1111 bp到-601 bp;在该区间内存在着参与水杨酸反应的顺式作用元件、对厌氧诱导必不可少的顺式调节元件以及参与干旱诱导的MYB结合位点,说明该区间可能存在着一些转录因子在上游调控BrGOLDEN基因的表达,从而导致不同类型的大白菜内叶类胡萝卜素的差异积累。这些发现为进一步探究BrGOLDEN的分子调控机制提供了一定依据。Both golden inner Chinese cabbage and orange inner Chinese cabbage are rich in carotenoids,with bright colors that are widely popular.Therefore,inner leaf color has become one of the important target traits for the breeding of new Chinese cabbage varieties and its regulatory mechanism is still unclear.A backcross population was constructed using golden inner and orange inner Chinese cabbages,and the molecular markers of golden inner trait of BrGOLDEN gene and the orange inner trait of Bror gene were developed to analyze the interaction between BrGOLDEN and Bror.RNA sequencing(RNA-seq)were used analyzed to the inner leaves of golden inner and light golden inner Chinese cabbage at mature stage to understand the expression of genes related to carotenoid metabolism pathways.Key cis-acting elements of the BrGOLDEN promoter were cloned and analyzed,and the core functional region of the BrGOLDEN promoter was explored using 5'truncated promoter cloning and GUS staining.The combination of genetic laws and molecular marker results indicated that the dominant gene BrGOLDEN and the recessive gene Bror were not inherited independently,and the Bror exhibited an epistatic effect on the BrGOLDEN,meaning that when the BrGOLDEN and Bror coexisted,only the orange inner phenotype was displayed.Transcriptome data showed that the expression levels of related genes of 2-C-methyl-D-erythritol 4-phosphate(MEP)precursor synthesis pathway and carotenoid synthesis pathway in golden inner Chinese cabbage were generally higher than those in light golden inner Chinese cabbage.The expression levels of genes involved in the synthesis of PSY and ZEP in the carotenoid synthesis pathway were generally higher than those of other genes in the pathway,indicating that the BrGOLDEN may control the synthesis of more carotenoids in Chinese cabbage by regulating the start and end of the related genes of carotenoid synthesis pathway.The analysis of promoter function showed that promoters with different deletion lengths exhibited different performance in ter

关 键 词：大白菜 类胡萝卜素 金黄心 橘红心 调控机制 遗传规律 分子标记 生物育种 

分 类 号：S63[农业科学—蔬菜学]