Fig. 8: OsTSN1 regulates the mRNA expression of OsJAZ6, OsJAZ12 and OsATG8C via degrading the mRNAs of OsNAC15 and OsLHY to promote SRBSDV infection.
a–i Phenotypes of uninfected or SRBSDV-infected WT, osjaz6, osjaz12, and osatg8c knockout mutant rice plants at 30 dpi (a–c). Scale bar, 10 cm. RT-qPCR results showing the expression level of SRBSDV P10 gene in SRBSDV-infected WT and osjaz6, osjaz12, osatg8c knockout mutant plants at 30 dpi (d–f). The data are the means ± SD from three independent biological replicates, determined using the one-way ANOVA with Tukey’s test. **P < 0.01, ***P < 0.001, ***P < 0.0001. Western blot results showing the accumulation levels of SRBSDV P10 in uninfected and SRBSDV-infected WT, osjaz6, osjaz12, and osatg8c knockout mutant rice plants (g–i). The Rubisco bands are used to show equal loading. j In vitro assay showing the co-localization of Cy5-labeled OsNAC15 or OsLHY mRNAs with the OsTSN1-P6 droplets. OsUBQ5 mRNA was used as a control. Scale bar, 10 μm. k OsNAC15 and OsLHY mRNAs were degraded in vitro by recombinant OsTSN1. Different amounts of recombinant GST-P6 were incubated with recombinant OsTSN1 for 2 h before the addition of OsNAC15 or OsLHY mRNAs. GST-C-P6Q42A,Y57A was used as a negative control. Degradation of OsNAC15 or OsLHY mRNAs was determined by nucleic acid electrophoresis. l, m OsTSN1 regulates the transcriptional levels of OsJAZ6, OsJAZ12 and OsATG8C through degrading mRNAs of OsLHY and OsNAC15. Images were taken at 2 dpa (l). The LUC/REN ratio represents the relative LUC activity (m). The data are the means ± SD from three independent biological replicates, determined using two-tailed Student’s t test. *P < 0.05, **P < 0.01. Experiments in (g–l) were repeated three times with similar results.
