Rice grassy stunt virus p5 interacts with two protein components of the plant-specific CBL–CIPK Ca+2 signaling network of rice

Rice grassy stunt virus (RGSV) is a tenuivirus posing a threat to rice production in many South, Southeast, and East Asian countries. To date, no host factor interacting with RGSV has been reported. In this study, we screened a rice cDNA library with the GAL4-based yeast two-hybrid system using RGSV p5 as the bait. One of the candidate host factors interacting with RGSV p5 was found to be CBL-interacting protein kinase 25 (OsCIPK25), a member of the plant-specific CBL-CIPK Ca2+ signaling network. The interaction between RGSV p5 and OsCIPK25, as well as OsCIPK5, which is closely related to OsCIPK25, was confirmed by their cellular co-localization and by a bimolecular fluorescence complementation assay in Nicotiana benthamiana cells. Given the importance of CIPKs in the regulation of ion homeostasis and the resemblance of RGSV symptoms to potassium deficiency in rice, we evaluated potassium content of RGSV-infected rice and found it to be much lower than that in the healthy rice.