Well-conserved mitogen-activated protein kinase (MAPK) cascades are essential for orchestrating of a wide range of cellular processes in plants, including defense responses against pathogen attack. NAC transcription factors (TFs) play important roles in plant immunity, but their targets and how they are regulated remain largely unknown. Here, we identified the TF OsNAC29 as a key component of a MAPK signaling pathway involved in rice (Oryza sativa) disease resistance. OsNAC29 binds directly to CACGTG motifs in the promoters of OsTPS28 and OsCYP71Z2, which are crucial for the biosynthesis of the phytoalexin 5,10-diketo-casbene and consequently rice blast resistance. OsNAC29 positively regulates rice blast resistance by promoting the expression of of OsTPS28 and OsCYP71Z2, and the function of OsNAC29 is genetically dependent on OsCYP71Z2 and OsTPS28. Furthermore, OsNAC29 interacts with OsRACK1A and OsMAPK3/6 to form an immune complex; OsMAPK3 phosphorylates OsNAC29 at Thr304 to prevent its proteasome-mediated degradation and promote its function against rice blast fungus. Phosphorylation of OsNAC29 at Thr304 is induced upon Magnaporthe oryzae infection and chitin treatment. Our data demonstrate the positive role of the OsMAPK3–OsNAC29–OsTPS28/OsCYP71Z2 module in rice blast resistance, providing insights into the molecular regulatory network and fine-tuning of NAC TFs in rice immunity.

Plants need to fine‐tune defense responses to maintain a robust but flexible host barrier to various pathogens. Helix‐loop‐helix/basic helix‐loop‐helix (HLH/bHLH) complexes play important roles in fine‐tuning plant development. However, the function of these genes in plant immunity and how they are regulated remain obscure. Here, we identified an atypical bHLH transcription factor, Oryza sativa (Os)HLH46, that interacts with rice receptor‐like cytoplasmic kinase (RLCK) Os BRASSINOSTEROID‐SIGNALING KINASE1‐2 (OsBSK1‐2), which plays a key role in rice blast resistance. OsBSK1‐2 stabilized OsHLH46 both in vivo and in vitro. In addition, OsHLH46 positively regulates rice blast resistance, which depends on OsBSK1‐2. OsHLH46 has no transcriptional activation activity and interacts with a typical bHLH protein, OsbHLH6, which negatively regulates rice blast resistance. OsbHLH6 binds to the promoter of OsWRKY45 and inhibits its expression, while OsHLH46 suppresses the function of OsbHLH6 by blocking its DNA binding and transcriptional inhibition of OsWRKY45. Consistent with these findings, OsWRKY45 was up‐regulated in OsHLH46‐overexpressing plants. In addition, the oshlh46 mutant overexpressing OsbHLH6 is more susceptible to Magnaporthe oryzae than is the wild type, suggesting that OsHLH46 suppresses OsbHLH6‐mediated rice blast resistance. Our results not only demonstrated that OsBSK1‐2 regulates rice blast resistance via the OsHLH46/OsbHLH6 complex, but also uncovered a new mechanism for plants to fine‐tune plant immunity by regulating the HLH/bHLH complex via RLCKs.

Plant nucleotide-binding leucine-rich repeat receptors (NLRs) sense pathogen effectors and activate effector-triggered immunity (ETI). Many plant NLRs form pairs with other NLRs to recognize effectors and initiate ETI. PIRICULARIA ORYZAE RESISTANCE IN BL1 (Pib), an NLR protein in rice (Oryza sativa), activates resistance by recognizing the rice blast effector AvrPib. The activation of Pib is suppressed by SH3 DOMAIN-CONTAINING PROTEIN 2 (OsSH3P2) in the absence of AvrPib. However, how Pib triggers defense responses and whether Pib pairs with another NLR are not clear. In this study, we identified Pib by map-based cloning and showed that a homolog of Pib, PIB HOMOLOGUE 8 (PibH8), interacts with Pib. Pib and PibH8 mediate resistance to the Magnaporthe oryzae isolate Guy11, a rice blast strain carrying AvrPib. Interestingly, the pib/pibh8 double mutant exhibited enhanced susceptibility to Guy11 compared to the single mutant. Furthermore, PibH8 can oligomerize through its coiled-coil (CC) domain, which also contributes to the Pib–PibH8 interaction, suggesting that Pib and PibH8 may form a complex to recognize AvrPib. OsSH3P2 inhibited the interaction of Pib and PibH8 through association with the CC domain of PibH8. Taken together, these results indicate that both Pib and PibH8 are required for rice blast resistance to M. oryzae carrying AvrPib, which is negatively regulated by OsSH3P2. This study not only identifies an NLR that functions in rice blast resistance but also reveals a possible complex immune strategy in which homologous NLR proteins may regulate the complete activation of plant immunity.