木薯MeHsfB3a互作蛋白的筛选和鉴定

Screening and Identification of MeHsfB3a Interacting Proteins in Cassava

热激蛋白转录因子Me Hsf B3a参与调控木薯对细菌性枯萎病的抗性,然而该蛋白不具有热激蛋白转录因子典型的转录激活结构域,可能与其他转录因子协同调控下游抗病基因表达。本研究构建了pGBKT7-MeHsfB3a载体,通过自激活实验证明Me Hsf B3a不具有转录激活活性,通过筛选Xpm HN11病原菌侵染的SC8木薯cDNA文库,获得Me Hsf B3a的候选互作蛋白MeMYB268,该蛋白属于MYB转录因子基因家族。克隆获得MeMYB268基因的编码区,全长936 bp,编码311个氨基酸。利用酵母点对点杂交进一步验证了MeHsfB3a与MeMYB268的互作关系。分析NCBI数据库中Xpm病原菌侵染木薯叶片0、5、7 d后的转录组数据,发现MeMYB268基因的表达受病原菌侵染诱导,在侵染7 d后该基因表达为对照的11倍。表明MeMYB268参与木薯对细菌性枯萎病的响应过程。本研究筛选获得了MeHsfB3a的互作蛋白MeMYB268,有助于进一步解析MeHsfB3a调控木薯对细菌性枯萎病的抗病机理。

Heat shock protein transcription factor MeHsfB3a is involved in regulating the resistance of cassava to cassava bacterial blight.However,this protein does not have the typical transcriptional activation domain of heat shock protein transcription factors,and may coordinate with other transcription factors to regulate the expression of downstream resistance genes.In this study,the pGBKT7-MeHsfB3a vector was constructed,and the self-activation experiment proved that MeHsfB3a does not have transcriptional activation activity.By screening the SC8 cassava cDNA library infected by XpmHN11 pathogen,the candidate interaction protein MeMYB268 of MeHsfB3a was obtained,which belongs to the MYB transcription factor gene family.The coding region of the MeMYB268 gene was cloned,with a ful length of 936 bp and encoding 311 amino acids.The interaction between Me HsfB3a and MeMYB268 was verified by yeast point-to-point hybridization.The transcriptome data of cassava leaves infected by Xpm pathogenic bacteria at O0,5 and 7 d in NCBI database were analyzed,and it was found that the expression of MeMYB268 gene was induced by pathogen infection,and the expression of this gene was 11 times higher than that of the control after 7 d of infection.It indicated that MeMYB268 was involved in the response of cassava bacterial blight.In this study,the interacting protein MeMYB268 of MeHsfB3a was screened,which is helpful to further analyze the mechanism of MeHsfB3a regulating cassava resistance to cassava bacterial blight.