紫心甘薯转录因子IbMYB1互作蛋白的筛选与验证  

Screening and Verification of Transcription Factor IbMYB1-Interacting Proteins in Purple Sweet Potato

作　　者：杨少华 陈亚慧 付丹文 高峰 Yang Shaohua;Chen Yahui;Fu Danwen;Gao Feng(Guangdong Provincial Key Laboratory of Biotechnology for Plant Development,School of Life Sciences,South China Normal University,Guangzhou,510631)

机构地区：[1]华南师范大学生命科学学院,广东省植物发育生物工程重点实验室,广州510631

出　　处：《分子植物育种》2024年第13期4192-4202,共11页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31171601);广东省重点领域研发计划项目(2020B20219001)共同资助。

摘　　要：Ib MYB1是调控紫心甘薯块根花色素苷生物合成的重要转录因子。为了鉴定与转录因子IbMYB1相互作用且参与花色素苷生物合成与调控的新蛋白,本研究通过酵母双杂交文库筛选其互作蛋白,获得了91个潜在互作蛋白的基因序列。通过对这些基因序列进行功能注释和统计分析,初步确定5个可能与花色素苷合成相关的候选基因,分别为IbCPK、IbMPK3、IbCSN3、IbFbox-5和IbMPK4。采用酵母双杂交回转实验和双分子荧光互补实验对候选蛋白与IbMYB1间的互作关系进行了再验证,最终确定其中IbCSN3和IbMPK4是与Ib MYB1互作且可能参与花色素苷生物合成调控的互作蛋白。本研究为鉴定IbMYB1的互作蛋白提供了实验参考,为丰富紫心甘薯花色素苷生物合成与调控的分子网络提供了新的线索。IbMYB1 is an important transcription factor that regulates anthocyanin biosynthesis in storage roots of purple sweet potato.In order to identify new proteins that interact with transcription factor IbMYB1 and participate in anthocyanin biosynthesis and regulation,in this study,we screened the interacting proteins through yeast two-hybrid library,and obtained the gene sequences of 91 potential interacting proteins.According to the GO annotation and statistical analysis of the selected genes,5 candidate genes that may be related to anthocyanin biosynthesis were preliminarily determined,namely IbCPK,IbMPK3,IbCSN3,IbFbox-5 and IbMPK4.The interaction between the candidate protein and IbMYB1 was re-verified by the yeast two-hybrid rotation experiment and the bimolecular fluorescence complementation experiment.It was finally determined that IbCSN3 and IbMPK4 were interacting proteins that interact with IbMYB1 and may participate in the regulation of anthocyanin biosynthesis.This study provides an experimental reference for the identification of IbMYB1 interaction proteins,and provides new clues for enriching the molecular network of anthocyanin biosynthesis and regulation in purple sweet potato.

关 键 词：紫心甘薯 转录因子 IbMYB1 酵母双杂交 蛋白互作 

分 类 号：S531[农业科学—作物学]