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

RCY1编码一种cc - nb - lrr类抗性蛋白，可使拟南芥对黄瓜花叶病毒（CMV(Y)）黄色株产生抗性。为了了解RCY1介导的病毒防御反应以及生物和非生物胁迫反应的串扰，以RCY1的CC-NB结构域为诱饵，在酵母双杂交系统中分离出与RCY1相互作用的宿主蛋白。结果，WRKY70转录因子被分离为RCY1相互作用的候选因子。通过共免疫纯化证实了WRKY70与CC-NB结构域的相互作用，但不是完整蛋白。在RCY1-HA转化的wrky70-1突变体的CMV(Y)接种叶片中，与携带RCY1-HA的野生型植株相比，病毒外壳蛋白积累增加，PR-1表达的诱导受到CMV(Y)的部分损害。生物和非生物胁迫的信号可能通过RCY1-WRKY70相互作用连接，因为WRKY70在两者中都起重要作用。

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.