大葱类黄酮3-O-葡萄糖基转移酶AfUFGT基因的克隆与表达分析  

Cloning and Expression Analysis of Flavonoid-3-O-Glucosyltransferase AfUFGT from Welsh Onion

作　　者：徐欢欢 李逸 邢嘉怡 王永勤[2] 刘乐承[1] Xu Huanhuan;Li Yi;Xing Jiayi;Wang Yongqin;Liu Lecheng(College of Horticulture and Gardening,Yangtze University,Jingzhou,434025;Key Laboratory of Horticultural Crop Biology and Germplasm Creation in North China(Ministry of Agriculture and Rural Areas)Vegetable Research Center,Beijing Academy of Agriculture and Forestry,Beijing,100097;Key Laboratory of Special Fruits and Vegetables Cultivation Physiology and Germplasm Resources Utilization,Xinjiang Production and Construction Group,Agricultural College of Shihezi University,Shihezi,832003)

机构地区：[1]长江大学园艺园林学院,荆州434025 [2]北京市农林科学院蔬菜研究中心,农业农村部华北地区园艺作物生物学与种质创制重点实验室,北京100097 [3]石河子大学农学院,新疆生产建设兵团特色果蔬栽培生理与种质资源利用重点实验室,石河子832003

出　　处：《分子植物育种》2024年第23期7678-7683,共6页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31972409);北京市农林科学院蔬菜研究中心改革与发展项目(KYCX10);北京市农林科学院科技创新能力项目(KJCX20200113)共同资助。

摘　　要：类黄酮3-O-葡萄糖基转移酶UFGT是类黄酮合成途径中最后的一个酶,在植物类黄酮的合成中具有重要作用。本研究以大葱为材料,根据大葱转录组序列设计特异性引物,使用RT-PCR从中获得编码大葱3-类黄酮葡萄糖基转移酶的基因,命名为AfUFGT。生物信息学分析及组织表达分析结果表明,AfUFGT编码区全长1425 bp,编码475个氨基酸,相对分子质量为52.31 kD,对应蛋白为正电荷的亲水性蛋白。进化分析表明AfUFGT与葱属植物洋葱AcUFGT2亲缘关系最近。亚细胞定位预测结果显示,AfUFGT编码的蛋白质定位于细胞质中。蛋白质序列多重比对结果显示,UFGT在多个物种中保守性较低,但与其他UFGT都具有完整的糖基转移酶的保守结构域。荧光定量PCR结果显示AfUFGT主要在根与叶片中表达。本研究结果为进一步阐明AfUFGT调控大葱类黄酮生物合成机制提供了理论基础。Flavonoid-3-O-glucosyltransferase(UFGT)is the last enzyme in the flavonoid synthesis pathway.It plays an important role in the synthesis of plant flavonoids.In this study,the specific primers were designed according to the transcriptome sequence of Welsh onion,UFGT was obtained by RT-PCR and named AfUFGT.Bioinformatics analysis showed that the total length of AfUFGT coding region was 1425 bp,encoding 475 amino acids,the relative molecular weight was 52.31 kD,and the corresponding protein was a positively charged hydrophilic protein.Evolutionary analysis showed that it was closely related to onion AcUFGT2.The subcellular localization prediction showed that the protein encoded by AfUFGT was localized in cytoplasm.Multiple alignments of protein sequences showed that UFGT was less conservative in many species,but it had a complete conserved domain of glycosyltransferase compared with other UFGT.Real-time qPCR showed that AfUFGT was mainly expressed in roots and leaves.The results of this study provide a theoretical basis for further clarifying the mechanism of AfUFGT regulating flavonoids biosynthesis in Welsh onion.

关 键 词：大葱 类黄酮3-O-葡萄糖基转移酶 表达分析 生物信息学分析 

分 类 号：S633.1[农业科学—蔬菜学]