Fig. 5: P2 attenuates OsChtBL1-mediated resistance to SBPH and RSV.
a The protein degradation assay showing P2 attenuates the protein stability of OsChtBL1 conferred by chitin. Water or 10 μg/ml Chitin and Purified GST or GST-P2 were added respectively to the total protein extract of OsChtBL1-OE plants, treated with 100 μg/mL CHX, and DMSO or MG132, Samples were collected every 30 min at 28 °C and analyzed by western blot using anti-MYC antibody. b The quantity of honeydew collected from NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX and P2-OX plants after SBPH feeding for 48 h. n = 19 individual SBPH. c The mortality rate of SBPH fed with NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX, and P2-OX plants after 15 days. d Phenotypes of Mock- or RSV-infected NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX, and P2-OX plants after RSV inoculation for 30 days. Scale bar, 10 cm. e The incidence rate of NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX, and P2-OX plants under field conditions at 30 days. f RT-qPCR assay showing the mRNA relative expression level of RSV CP in RSV-infected NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX, and P2-OX plants. g Western blot assay showing protein accumulation level of RSV CP in Mock- or RSV-infected NIP, OsChtBL1-OE#1, OsChtBL1-OE#1/P2-OX, and P2-OX plants. OsUBQ5 serves as an internal reference gene (f). n = 3 (b, c, e, f) 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, and ns at the top of columns indicates no 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 (a, g). Experiments in (a, g) were repeated three times with similar results.
