摘要
TaCRK2在叶锈菌侵染小麦诱发的过敏性反应中起正调控作用,为深入了解TaCRK2在小麦抵抗叶锈菌侵染中的作用机制,本试验采用串联亲和纯化与质谱联用(TAP-MS)技术,鉴定出翻译控制肿瘤蛋白(Translationally controlled tumor protein,TCTP)为TaCRK2的潜在互作蛋白。但TaCRK2与TaTCTP的相互作用还缺乏试验证据,本研究利用DUALmembrane系统酵母双杂交(DUALmembrane system yeast-twohybrid)和双分子荧光互补(Bimolecular fluorescence complementation,BiFC)技术验证TaCRK2和TaTCTP之间的相互作用。DUALmembrane系统酵母双杂交结果显示,同时携带TaCRK2和TaTCTP的酵母NMY51可以在缺陷型培养基上生长;BiFC试验结果显示,TaCRK2和TaTCTP发生相互作用,并且定位在烟草表皮细胞的内质网膜上。本研究为深入了解TaCRK2在叶锈菌侵染小麦诱发的过敏性反应中的作用机制奠定了基础。
TaCRK2 plays a positive regulation role in the hypersensitive reaction induced by Puccinia triticina infection of wheat.To further understand the role of TaCRK2 resistance to Puccinia triticina infection,the translationally controlled tumor protein(TCTP)was identified as a potential interaction protein of TaCRK2 by using tandem affinity purification and mass spectrometry(TAP-MS)technology.However,there is still no experimental evidence for the interaction between TaCRK2 and TaTCTP.In this study,DUALmembrane system yeast two-hybrid and BiFC experiments were used to verify the interaction between TaCRK2 and TaTCTP.Through the DUALmembrane system yeast two-hybrid,it was found that yeast NMY51 with both TaCRK2 and TaTCTP could grow on a defective medium.By BiFC experiments,it was found that the fluorescence signal of the interaction between TaCRK2 and TaTCTP occurred on the endoplasmic reticulum membrane of tobacco epidermal cells.This study lays the foundation for a deep understanding of the mechanism of TaCRK2 in the hypersensitive reaction induced by Puccinia triticina infection of wheat.
作者
顾佳
李浩
安瑞朋
刘刚
王冬梅
GU Jia;LI Hao;AN Ruipeng;LIU Gang;WANG Dongmei(State Key Laboratory of North China Crop Improvement and Regulation,Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology/College of Life Sciences,Hebei Agricultural University,Baoding 071001,China)
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2020年第4期16-22,共7页
Journal of Hebei Agricultural University
基金
国家自然科学基金(31171472
31871548)
高等学校博士学科点专项科研基金资助课题(优先发展领域)(20111302130001)
河北省应用基础研究计划重点基础研究项目(12967149D).
