OsWRKY10 extensively activates multiple rice diterpenoid phytoalexin biosynthesis to enhance rice blast resistance.

Phytoalexin is the main chemical weapon against disease pathogens in plants. Rice produces a number of phytoalexins to defend pathogens, most of which belong to diterpenoid phytoalexins. Three biosynthetic gene clusters (BGCs) and a few non-cluster genes are responsible for rice diterpenoid phytoalexin biosynthesis. The corresponding regulatory mechanism of these phytoalexins in response to pathogen challenges still remains unclear. Here we identified a transcription factor, OsWRKY10, positively regulating rice diterpenoid phytoalexin biosynthesis. Knockout mutants of OsWRKY10 obtained by the CRISPR/Cas9 technology are more susceptible to Magnaporthe oryzae infection, while overexpression of OsWRKY10 enhances resistance to rice blast. Further analysis reveals that overexpression of OsWRKY10 increases accumulation of multiple rice diterpenoid phytoalexins and expression of genes in three BGCs and non-clustered genes in response to M. oryzae infection. Knockout of OsWRKY10 impairs upregulation of rice diterpenoid phytoalexin biosynthesis gene expression by blast pathogen and CuCl 2 treatment. OsWRKY10 directly binds to the W-boxes or W-box-like elements (WLEs) of rice diterpenoid phytoalexin biosynthesis gene promoters to regulate the corresponding gene expression. This study identified an extensive regulator (OsWRKY10) with the broad transcriptional regulation on rice diterpenoid phytoalexin biosynthesis, providing the insight to characterize regulation of rice chemical defense for improving disease resistance.