丹参3个R2R3-MYB转录因子的亚细胞定位和自激活活性分析

摘要

R2R3-MYB转录因子能够有效调控丹参酚酸类物质的积累.将丹参的SmMYB33、SmMYB54和SmMYB93与拟南芥的R2R3-MYB进行比对,找到拟南芥中与丹参3个转录因子亲缘关系最近的功能已知的转录因子,根据这些功能已知的转录因子,对丹参3个转录因子的功能进行预测.构建含有绿色荧光报告基因的载体pA7-GFP-SmMYB33、pA7-GFP-SmMYB54和pA7-GFP-SmMYB93,使用基因枪方法将载体分别转化洋葱表皮细胞进行亚细胞定位分析,结果表明3个转录因子主要定位在细胞核,绿色荧光也有少部分出现在细胞质中.构建酵母表达载体pDEST-GBKT7-SmMYB33、pDEST-GBKT7-SmMYB54和pDEST-GBKT7-SmMYB93用于酵母自激活活性分析,结果表明SmMYB33有自激活活性,SmMYB54和SmMYB93没有自激活活性.%R2R3-MYB can regulate accumulation of phenolic acids efficiently in Salvia miltiorrhiza Bunge.In this study,the amino acid sequences of SmMYB33,SmMYB54,SmMYB93 and R2R3-MYBs of Arabidopsis thaliana were aligned to find the R2R3-MYBs of Arabidopsis thaliana which had the closest relationships with the three MYBs of S.miltiorrhiza.According to the phylogenetic analysis,the functions of three MYBs were predicted.The plasmids pA7-GFP-SmMYB36,pA7-GFP-SmMYB54 and pA7-GFP-SmMYB93 were constructed and transiently transformed into onion epidermis to conduct subcellular localization analysis by using gene gun.The results showed that three MYBs were mainly localized in the nucleus of onion epidermis,and a small amount of the GFP fluorescence also appeared in the cytoplasm.The plasmids pDEST-GBKT7-SmMYB36,pDEST-GBKT7-SmMYB54 and pDEST-GBKT7-SmMYB93 were constructed for transactivation analysis.The results implied that SmMYB54 and SmMYB93 did not have the transactivation activity,and SmMYB33 had the transactivation activity.