Fig. 6: P2 facilitates the degradation of OsChtBL1 mediated by OsRING18.

a, c Y2H assay showing OsRING18 interacts with OsChtBL1 and P2 in yeast cells. These yeast cells containing different combinations were observed after growing on the SD-LT and SD-AHLT medium at 30 °C for 3 days. b, d Co-IP assay showing OsRING18 interacts with OsChtBL1 and P2 in N. benthamiana. e The protein degradation assay showing P2 synergizes with OsRING18 to promote the degradation of OsChtBL1. f The protein degradation assay showing the degradation of OsChtBL1 mediated by P2 is dependent on OsRING18. g The quantity of honeydew collected from ZH11 and osring18 plants after SBPH feeding for 48 h. n = 17 individual SBPH. h Phenotypes of Mock- or RSV-infected ZH11 and osring18 plants after RSV inoculation for 30 days. Scale bar, 10 cm. i The incidence rate of ZH11 and osring18 plants under field conditions at 30 days. j RT-qPCR assay showing the mRNA relative expression level of RSV CP in RSV-infected ZH11 and osring18 plants. k Western blot assay showing protein accumulation level of RSV CP in Mock- or RSV-infected ZH11 and osring18 plants. OsUBQ5 serves as an internal reference gene (j). n = 3 (g, i, j) independent biological replicates. Error bars represent SD, and values are means ± SD. All the statistical analysis data were performed using a two-tailed Student’s t test. *At the top of columns indicates significant differences with the control group at P < 0.05. Rubisco serves as an internal reference protein for Western blot analysis and detected by anti-RUB antibody (e, f, k). Experiments in (b, d–f, k) were repeated three times with similar results.