TaJAZ7D蛋白在冬小麦JA抗寒途径中的作用

Role of TaJAZ7D Protein in JA Cold-Resistance Pathway of Winter Wheat

转录抑制因子JAZ(Jasmonate ZIM-domain)蛋白是茉莉酸(Jasmonate,JA)信号转导途径的核心元件,前人研究发现,拟南芥中AtJAZ1、AtJAZ4与AtICE1蛋白互作可抑制AtICE1基因的转录,负调控拟南芥的抗寒性,然而小麦中TaJAZ与TaICE蛋白的互作调控机制尚不清楚。本研究以强抗寒冬小麦品种东农冬麦1号(Dn1)为试验材料,以与拟南芥AtJAZ1、AtJAZ4蛋白高度同源的TaJAZ7、TaJAZ12蛋白为对象,检测外源茉莉酸甲酯(MeJA)对低温胁迫下TaJAZ7、TaJAZ12基因表达量的影响。结果发现,TaJAZ7基因的表达量与温度变化呈明显负相关,故对TaJAZ7(以TaJAZ7D为研究对象进行后续研究)蛋白进行生物信息学分析和亚细胞定位,并利用酵母双杂交技术验证TaJAZ7D蛋白与TaMYC2和TaICE41蛋白的互作。生物信息学分析表明,TaJAZ7D基因编码区序列全长为633bp,为不稳定的亲水性混合型蛋白;TaJAZ7D蛋白有典型的TIFY和Jas结构域,属于TIFY家族。亚细胞定位发现,TaJAZ7D蛋白位于细胞核中。酵母双杂交验证试验发现,TaJAZ7D蛋白与TaMYC2和TaICE41蛋白分别存在互作关系。

The transcriptional repressor protein JAZ(Jasmonate ZIM-domain)is a core component of JA(Jasmonate)signaling pathway.AtJAZ1,AtJAZ4 and AtICE1 proteins interact to repress AtICE1 gene transcription,and thus negatively regulate cold resistance in Arabidopsis.However,it is not clear whether TaJAZ protein is involved in the regulation of cold resistance in wheat by interacting with TaICE protein.In this work,TaJAZ7 and TaJAZ12 proteins,which are highly homologous to AtJAZ1 and AtJAZ4 were used as experimental factors,and the strong cold-resistant winter wheat variety Dongnongdongmai 1(Dn1)was used as the experimental material,to examine the effect of exogenous MeJA treatment on the expression of TaJAZ7 and TaJAZ12 genes under low temperature stress.The results showed that the changes of TaJAZ7 gene expression level were significantly negatively regulated with decreasing temperature.Bioinformatics analysis and subcellular localization TaJAZ7 protein(TaJAZ7 D protein was used as research object for follow-up studies)and were performed.And the interaction between TaJAZ7 D and TaMYC2,TaICE41 proteins was studied through yeast two-hybrid system.Bioinformatics analysis indicated that TaJAZ7 D gene is a 633 bp in length,the encoding protein was unstable,hydrophilic,mixed-type protein,which contains the typical TIFY and Jas domain and belongs to the TIFY family.Subcellular localization analysis indicated TaJAZ7 D protein was localized in the nucleus.Yeast two-hybrid assay verified that TaJAZ7 D protein interacts with TaMYC2 and TaICE41 proteins,respectively.