Abnormally developed endosperm strongly affects rice (Oryza sativa) appearance quality and grain weight. Endosperm formation is a complex process, and although many enzymes and related regulators have been identified, many other related factors remain largely unknown. Here, we report the isolation and characterization of a recessive mutation of White Belly 1 (WB1), which regulates rice endosperm development, using a modified MutMap method in the rice mutant wb1. The wb1 mutant develops a white-belly endosperm and abnormal starch granules in the inner portion of white grains. Representative of the white-belly phenotype, grains of wb1 showed a higher grain chalkiness rate and degree and a lower 1000-grain weight (decreased by ~34%), in comparison with that of Wild Type (WT). The contents of amylose and amylopectin in wb1 significantly decreased, and its physical properties were also altered. We adopted the modified MutMap method to identify 2.52 Mb candidate regions with a high specificity, where we detected 275 SNPs in chromosome 4. Finally, we identified 19 SNPs at 12 candidate genes. Transcript levels analysis of all candidate genes showed that WB1 (Os04t0413500), encoding a cell-wall invertase, was the most probable cause of white-belly endosperm phenotype. Switching off WB1 with the CRISPR/cas9 system in Japonica cv. Nipponbare demonstrates that WB1 regulates endosperm development and that different mutations of WB1 disrupt its biological function. All of these results taken together suggest that the wb1 mutant is controlled by the mutation of WB1, and that the modified MutMap method is feasible to identify mutant genes, and could promote genetic improvement in rice.

CuO NPs (cupric oxide nanoparticles) are widely used in various fields due to their high electrical conductivity, electronic correlation effect, and special physical property. Notably, CuO NPs have good application prospects in agricultural production because of its antifungal activity to prevent crop diseases. However, the increasing release of CuO NPs into the environment has resulted in a serious threat to the ecosystem, including plants. Previous studies have reported the toxicity of CuO NPs on rice, but little is known about the underlying molecular mechanisms or specific genes involved in the response to CuO NPs. In this study, we found that the rice well-known receptor Chitin Elicitor Receptor Kinase 1 (OsCERK1), which is essential for basal resistance against pathogens, is involved in CuO NPs stress in rice. Knockout of OsCERK1 gene resulted in enhanced tolerance to CuO NPs stress. Furthermore, it was revealed that OsCERK1 reduces the tolerance to CuO NPs stress by regulating the anti-oxidant system and increasing the accumulation of H₂O₂ in rice. In addition, CuO NPs treatment significantly enhances the basal resistance against M. oryzae which is mediated by OsCERK1. In conclusion, this study demonstrated a dual role of OsCERK1 in response to CuO NPs stress and M. oryzae infection by modulating ROS accumulation, which expands our understanding about the crosstalk between abiotic and biotic stresses.

Phytophthora cinnamomi causes significant root rot in hickory, leading to substantial yield losses. While Bacillus spp. are recognized as beneficial rhizosphere microorganisms, their application against hickory root rot and their impact on rhizosphere microbial communities remain under-investigated. This study demonstrated that Bacillus velezensis TCS001 significantly inhibited P. cinnamomi ST402 growth in vitro, and achieved 71% efficacy in root rot disease management. Scanning electron microscopy (SEM) revealed that TCS001 fermentation filtrate induced mycelial deformities in P. cinnamomi. An analysis of α and β diversity indicated a significant impact of TCS001 on rhizosphere bacterial community richness and diversity, with minimal effects on the fungal community. Moreover, TCS001 altered the hickory rhizosphere microbiome co-occurrence network. The differential abundance analysis suggests that TCS001 promotes the recruitment of beneficial microbes associated with disease resistance, thereby suppressing disease development. These findings underscore the influence of TCS001 on the hickory rhizosphere microbiome in the presence of pathogens, providing valuable data for future research and the development of effective biocontrol strategies for hickory root rot.

Mitophagy refers to the specific process of degrading mitochondria, which is an important physiological process to maintain the balance of mitochondrial quantity and quality in cells. At present, the mechanisms of mitophagy in pathogenic fungi remain unclear. Magnaporthe oryzae (Syn. Pyricularia oryzae), the causal agent of rice blast disease, is responsible for the most serious disease of rice. In M. oryzae, mitophagy occurs in the foot cells and invasive hyphae to promote conidiation and infection. In this study, fluorescent observations and immunoblot analyses showed that general stress response protein MoWhi2 is required for mitophagy in M. oryzae. In addition, the activation of the autophagy, pexophagy and cytoplasm-to-vacuole targeting (CVT) pathway upon nitrogen starvation was determined using the GFP-MoATG8, GFP-SRL and MoAPE1-GFP strains and the ΔMowhi2 mutant in these backgrounds. The results indicated that MoWhi2 is specifically required for mitophagy in M. oryzae. Further studies showed that mitophagy in the foot cells and invasive hyphae of the ΔMowhi2 was interrupted, leading to reduced conidiation and virulence in the ΔMowhi2 mutant. Taken together, we found that MoWhi2 contributes to conidiation and invasive growth by regulating mitophagy in M. oryzae.

Rice blast, caused by the fungus Magnaporthe oryzae, is a highly damaging disease. Introducing genes, which confer a broad spectrum resistance to the disease, such as Pib, makes an important contribution to protecting rice production. However, little is known regarding the mechanistic basis of the products of such genes. In this study, transcriptome of the cultivar Lijiangxintuanheigu (LTH) and its monogenic IRBLb-B which harbors Pib treated with M. oryzae were compared. Among the many genes responding transcriptionally to infection were some encoding products involved in the metabolism of ROS (reactive oxygen species), in jasmonate (JA) metabolism, and WRKY transcription factors, receptor kinases, and resistance response signal modulation. The down-regulation of genes encoding peroxiredoxin and glutathione S transferases implied that the redox homeostasis is essential for the expression of Pib-mediated resistance. The up-regulation of seven disease resistance-related genes, including three encoding a NBS-LRR protein, indicated that disease resistance-related genes are likely tend to support the expression of Pib resistance. These data revealed that potential candidate genes and transcriptional reprogramming were involved in Pib-mediated resistance mechanisms.

Epigenetic modification is important for cellular functions. Trimethylation of histone H3 lysine 4 (H3K4me3), which associates with transcriptional activation, is one of the important epigenetic modifications. In this study, the biological functions of UvKmt2-mediated H3K4me3 modification were characterized in Ustilaginoidea virens, which is the causal agent of the false smut disease, one of the most destructive diseases in rice. Phenotypic analyses of the ΔUvkmt2 mutant revealed that UvKMT2 is necessary for growth, conidiation, secondary spore formation, and virulence in U. virens. Immunoblotting and chromatin immunoprecipitation assay followed by sequencing (ChIP-seq) showed that UvKMT2 is required for the establishment of H3K4me3, which covers 1729 genes of the genome in U. virens. Further RNA-seq analysis demonstrated that UvKmt2-mediated H3K4me3 acts as an important role in transcriptional activation. In particular, H3K4me3 modification involves in the transcriptional regulation of conidiation-related and pathogenic genes, including two important mitogen-activated protein kinases UvHOG1 and UvPMK1. The down-regulation of UvHOG1 and UvPMK1 genes may be one of the main reasons for the reduced pathogenicity and stresses adaptability of the ∆Uvkmt2 mutant. Overall, H3K4me3, established by histone methyltransferase UvKMT2, contributes to fungal development, secondary spore formation, virulence, and various stress responses through transcriptional regulation in U. virens.