CHS启动子差异片段在不同观赏海棠品种(系)中的功能验证  

Function assay of the difference of chalcone synthase gene promoter during Malus crabapples leaf coloration

作　　者：孙文静 许扬 高艺 张杰 宋婷婷 SUN Wenjing;XU Yang;GAO Yi;ZHANG Jie;SONG Tingting(College of Plant Science and Technology/Beijing Key Laboratory of New Technology in Agricultural Application/National Demonstration Center for Experimental Plant Production Education,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院植物科学技术学院/农业应用新技术北京市重点实验室/植物生产国家级实验教学示范中心,北京102206

出　　处：《北京农学院学报》2022年第3期49-54,共6页Journal of Beijing University of Agriculture

基　　金：国家林业和草原局林草国家创新联盟自筹研发项目(GLM[2020]17号)。

摘　　要：【目的】为探明查尔酮合酶基因的启动子序列差异在不同苹果属观赏海棠品种(系)中的功能。【方法】以‘王族’(Malus cv.‘Royalty’)和M10-5(Malus M10-5)及‘火焰’(Malus cv.‘Flame’)叶片为试材,克隆得到McCHS基因的启动子,并进行序列比对。检测McCHS基因在3个品种(系)中的表达水平,进一步通过酵母单杂交和凝胶阻滞迁移分析确定McCHS基因的启动子序列差异和调控花色素苷生物合成关键MYB转录因子McMYB10的结合关系。【结果】‘火焰’McCHS基因启动子与‘王族’McCHS基因启动子序列比对发现,‘火焰’McCHS基因启动子中有743 bp的缺失,而M10-5的McCHS基因启动子同时拥有缺失和完整的两种启动子序列;酵母单杂交和凝胶阻滞迁移结果表明,743 bp缺失序列能够和McMYB10结合,而且McCHS基因的表达量和花色素苷含量呈相同趋势。【结论】McCHS基因的启动子序列差异可能影响观赏海棠叶片着色。【Objective】The purpose of this experiment was to compare the effect of sequence differences of McCHS promoter region on leaf color in three cultivars of Malus crabapples.【Methods】The promoters of McCHS were cloned and compared the differences among Malus cv.‘Royalty’,Malus M10-5 and Malus cv.‘Flame’.The expression levels of McCHS were detected in three cultivars(line).The binding ability of anthocyanin biosynthesis regulator MYB with McCHS promoter was further determined by Yeast one hybrid(Y1H)and Electrophoretic mobility shift assay(EMSA).【Results】There is a 743 bp deletion in McCHS promoter region of Malus cv.‘Flame’compared to that of Malus cv.‘Royalty’.The McCHS promoter of Malus M10-5 has two different bands.The expression pattern of McCHS in the three cultivars(line)was similar to the trend of anthocyanin content in those three cultivars(line).McMYB10 can binds to the 743 bp deletion sequence by Y1H.【Conclusion】The sequence difference of McCHS promoter may involve in leaf coloration of Malus crabapples.

关 键 词：苹果属观赏海棠 查尔酮合酶基因 花色素苷 

分 类 号：S661.1[农业科学—果树学]