Magnaporthe oryzae is one of the most important fungal pathogens of rice. Chitin and avirulent strains can induce two layers of immunity response, pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI), in rice with cognate R genes. However, little is known about the assembly of the rice microbiome induced by PTI and ETI in rice. In this study, we investigate the impact of continuous treatment of the avirulent M. oryzae strain with AvrPi9 and chitin on the bacterial endophytic community of rice varieties harboring resistant gene Pi9 and their antagonistic activity against rice blast fungus. Analysis of the 16S rRNA showed a significant increase in the diversity and microbial co-occurrence network complexity and the number of beneficial taxa—Bacillus, Pseudomonas, Microbacterium, and Stenotrophomonas spp.—following the chitin and avirulent strain treatments. The antifungal assay with bacterial endophytes recovered from the leaves showed few bacteria with antagonistic potential in rice treated with avirulent strains, suggesting that the sequential treatment of the avirulent strain decreased the antagonistic bacteria against M. oryzae. Moreover, we identified Bacillus safensis Ch_66 and Bacillus altitudinis Nc_68 with overall antagonistic activities in vivo and in vitro. Our findings provide a novel insight into rice microbiome assembly in response to different innate immunity reactions.

Rice brown spot disease, caused by Bipolaris oryzae, is a significant fungal disease that poses a major threat to global rice production. Despite its widespread impact, genomic studies of B. oryzae remain limited, particularly those involving high-quality genomic data. In this study, we performed whole-genome sequencing of the B. oryzae strain RBD1, which was isolated from the demonstration field for upland rice cultivation in Haozhiba Village, Lancang County, Pu’er City, Yunnan Province, China, using a combination of second-generation Illumina sequencing and third-generation Single-Molecule Real-Time (SMRT) sequencing. The assembled genome was 37.5 Mb in size with a G + C content of 49.39%, containing 42 contigs with a contig N50 of 2.0 Mb. Genomic analysis identified genes related to carbon, nitrogen, and lipid metabolism, highlighting the strain’s metabolic flexibility under diverse environmental conditions and host interactions. Additionally, we identified pathogenicity-related genes involved in MAPK signaling, G protein signaling, and oxidative stress responses. Under 1.2 M sorbitol-induced osmotic stress, we observed significant differences in growth responses between RBD1 and the rice blast fungus Magnaporthe oryzae H7. Transcriptomic analysis using Illumina sequencing revealed that RBD1 responds to osmotic stress by enhancing carbohydrate metabolism, fatty acid degradation, and amino acid synthesis, while H7 primarily relies on protein synthesis to enhance growth tolerance. This study provides a valuable foundation for understanding the pathogenic mechanisms of rice brown spot and future disease control strategies.

Magnaporthe oryzae causes rice blast disease, which threatens global rice production. The interaction between M. oryzae and rice is regarded as a classic model for studying the relationship between the pathogen and the host. In this study, we found a gene, MoHG1, regulating fungal development and virulence in M. oryzae. The ∆Mohg1 mutants showed more sensitivity to cell wall integrity stressors and their cell wall is more easily degraded by enzymes. Moreover, a decreased content of chitin but higher contents of arabinose, sorbitol, lactose, rhamnose, and xylitol were found in the ∆Mohg1 mutant. Combined with transcriptomic results, many genes in MAPK and sugar metabolism pathways are significantly regulated in the ∆Mohg1 mutant. A hexokinase gene, MGG_00623 was downregulated in ∆Mohg1, according to transcriptome results. We overexpressed MGG_00623 in a ∆Mohg1 mutant. The results showed that fungal growth and chitin contents in MGG_00623-overexpressing strains were restored significantly compared to the ∆Mohg1 mutant. Furthermore, MoHG1 could interact with MGG_00623 directly through the yeast two-hybrid and BiFC. Overall, these results suggest that MoHG1 coordinating with hexokinase regulates fungal development and virulence by affecting chitin contents and cell wall integrity in M. oryzae, which provides a reference for studying the functions of MoHG1-like genes.