Screening of protein associated with crosstalk between disease-resistance and biotic/abiotic stress responses.

RCY 1编码CC-NB-LRR类抗性蛋白，其赋予对拟南芥中黄瓜花叶病毒[CMV（Y）]的黄色株系的抗性。为了了解RCY 1介导的病毒防御反应以及生物和非生物胁迫反应的串扰，使用RCY 1的CC-NB结构域作为诱饵，在酵母双杂交系统中分离与RCY 1相互作用的宿主蛋白。结果，WRKY 70转录因子被分离为RCY 1相互作用子的候选者。通过共免疫纯化证实了WRKY 70和CC-NB结构域之间的相互作用，但不是完整的蛋白质。与携带RCY 1-HA的野生型植物相比，在RCY 1-HA转化的wrky 70 -1突变体的CMV（Y）接种的叶中，病毒外壳蛋白积累增加，并且响应于CMV（Y）的PR-1表达的诱导部分受损。WRKY 70在生物胁迫和非生物胁迫信号转导中均发挥重要作用，因此RCY 1-WRKY 70可能是二者相互作用的信号转导途径。

RCY1 encodes a CC-NB-LRR-class resistance protein that confers resistance to a yellow strain of Cucumber mosaic virus [CMV(Y)] in Arabidopsis thaliana. To understand the RCY1-mediated virus defense response and crosstalk of biotic and abiotic stress responses, host protein that interacts with RCY1 was isolated in a yeast two-hybrid system using the CC-NB domains of RCY1 as bait. As the result, the WRKY70 transcription factor was isolated as a candidate of RCY1 interactor. The interaction between WRKY70 and the CC-NB domains, but not complete protein, was confirmed by co-immunopurification. In CMV(Y)-inoculated leaves of the RCY1-HA-transformed wrky70-1 mutant, virus coat protein accumulation increased and the induction of PR-1 expression in response to CMV(Y) was partially compromised as compared to wild-type plants carrying RCY1-HA. It is possible that signaling of biotic and abiotic stresses connect via RCY1-WRKY70 interaction since WRKY70 has important roles in both of them.