水稻水通道蛋白OsPIP1;3与白叶枯病菌harpin蛋白Hpa1互作关系研究  

Studies on the interaction between aquaporin OsPIP1;3 of rice and harpin protein Hpa1 of Xanthomonas oryzae pv. oryzae

作　　者：王欢[1] 田沂民 董汉松[1] 

机构地区：[1]南京农业大学植物保护学院,南京210095

出　　处：《植物生理学报》2016年第8期1223-1230,共8页Plant Physiology Journal

基　　金：公益性行业(农业)科研专项(201303015)~~

摘　　要：本文通过片段缺失和定点突变的方法构建产生水稻（Oryza sativa）水通道蛋白OsPIP1;3的变异本。通过膜酵母双杂交进行蛋白互作分析,实验结果表明水稻白叶枯病菌（Xanthomonas oryzae pv.oryzae）harpin蛋白Hpa1能与OsPIP1;3互作,但如果将OsPIP1;3的一个胞外序列（loopE）及OsPIP1;3的第六个跨膜区域（TM6）删除,则互作不能发生。进一步对276-279 aa进行替换,OsPIP1;3与Hpa1不再发生互作。这些结果说明,OsPIP1;3序列上的loop E和TM6上的氨基酸对OsPIP1;3–Hpa1互作有重要影响,而276-279 aa是可能的互作位点。In this study, we generated mutant versions of rice（Oryza sativa） aquaporin OsPIP1;3 by fragment deletion and site-directed mutagenesis, respectively. When a split-ubiquitin–based yeast two-hybrid system was employed to analyze protein combinations of Hpa1（a harpin protein of Xanthomonas oryzae pv. oryzae） with the canonical form and mutant versions of OsPIP1;3, interaction was found between Hpa1 and the canonical form, but not in the combination of Hpa1 with the mutant protein generated by deleting a short intracellular region called connecting loop E and transmembrane domain 6（TM6）. Analyses of additional substitutive protein mutants indicated that residues 276–279 in TM6 were also important for the molecular interaction. These data suggest that loop E and TM6 are determinants of OsPIP1;3 interaction with Hpa1 while particular residues 276–279 play crucial roles in the molecular interaction.

关 键 词：Hpa1 OsPIP1 3 定点突变 膜酵母双杂交 蛋白互作 

分 类 号：S435.111.47[农业科学—农业昆虫与害虫防治]