稻瘟病菌无毒效应因子Avr-PikD与水稻蛋白OsDjA9的互作鉴定

郭嘉媛; 洪永河; 黄健强; 吴亦灵; 王宗华; 陈松彪; 陈晓峰

doi:10.19303/j.issn.1008-0384.2022.005.015

稻瘟病菌无毒效应因子Avr-PikD与水稻蛋白OsDjA9的互作鉴定

Interaction Identification between Magnaporthe oryzae Avirulence Effector Avr-PikD and Rice Protein OsDjA9

摘要

摘要:
目的对从水稻cDNA文库中筛选出的一个候选互作蛋白OsDjA9与Avr-PikD的互作关系进行鉴定，以期获得稻瘟病菌无毒效应因子Avr-PikD的水稻靶标。
方法通过酵母双杂交、pull-down、Co-IP、荧光素酶互补成像试验及水稻原生质体中的共定位分析来验证Avr-PikD与OsDjA9的互作关系，并进一步借助酵母双杂交鉴定OsDjA9中参与互作的关键结构域。
结果 Avr-PikD在体外及体内条件下均能与OsDjA9发生相互作用，且OsDjA9所含DnaJ结构域是其与Avr-PikD互作所必需的。
结论在稻瘟病菌侵染水稻的过程中，水稻OsDjA9蛋白是稻瘟病菌无毒效应因子Avr-PikD的一个作用靶标。

Abstract:
Objective To obtain the rice target of Magnaporthe oryzae avirulence effector Avr-PikD, the interaction between Avr-PikD and OsDjA9, one of the candidate interacting proteins screened out from a rice cDNA library, was identified.
Method The interaction between Avr-PikD and OsDjA9 was verified by assays of yeast two-hybrid, pull-down, Co-IP, and luciferase complementation imaging along with co-localization analysis in the rice protoplasts. Specific domain in OsDjA9 involved in the interaction was determined by yeast two-hybrid assay.
Result Avr-PikD interacts with OsDjA9 in vitro as well as in vivo. The DnaJ domain of OsDjA9 is essential for the interaction.
Conclusion During the process of M. oryzae infecting rice, OsDjA9 is the target of the avirulence effector Avr-PikD secreted by the pathogen.

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