Rice Blast Research Articles

First Report of Burkholderia glumae Causing Bacterial Panicle Blight in Rice in Bangladesh.

Bacterial panicle blight (BPB) is one of the emerging diseases occurring in different Agro-Ecological Zones (AEZ) of Bangladesh and can cause up to 75% yield loss. In Bangladesh, the typical symptoms of BPB include sheath rot, panicle blight, grain spotting, and grain rot in both inbred and hybrid rice varieties, which resemble those reported by Zhou (2019). To confirm, 300 field samples of 20 panicles each with typical BPB symptoms from 20 districts (3 locations each district and 5 fields per location) were collected during mid-November 2022 for the causal pathogen(s) isolation. Nearly 70% of the panicles showed a dark brown chaffy appearance in the fields. For identification of the causal pathogen(s), 1 g of rice grains with typical BPB symptoms was surface sterilized by immersing for 15 seconds in 70% ethanol, 1 min in 3% sodium hypochlorite solution followed by rinsing the grains three times, and soaked in 1 mL sterile distilled water for 10 min (Mirghasempour et al. 2018). During grinding using mortar and pestle, 5 mL water was added (Islam et al. 2023) after which the suspension (20 μL) was then streaked onto the selective medium (S-PG) (Tsushima et al. 1986). Purple color colonies on the S-PG medium were selected and purified as candidate pathogens. For further confirmation, the genomic DNA of the bacterial isolates was extracted and amplified by PCR using 16SF (5'-AGAGTTTGATCCTGGCTCAG-3') and 16SR (5'-GGCTACCTTGTTACGACTT-3') (Nandakumar et al. 2009), and glu-FW (5'-GAAGTGTCGCCGATGGAG-3') and glu-RV (5'-CCTTCACCGACAGCACGCAT-3') primers (Maeda et al. 2006). The PCR products were visualized on 1% agarose gel resulting amplicons of 1494bp for 16S-rDNA and 529bp for gyrB. The PCR results revealed 529bp amplification for gyrB gene in one sample that was collected from a field in Natore (24°21'0.00" N 89°04'59.88" E) district cultivating Swarna (a local rice variety), primarily indicating the causal pathogen is Burkholderia glumae. The PCR products were sequenced using both primers and sequence data was analyzed by the BLAST nucleotide program. The obtained partial sequences of 16S rDNA and gyrB were deposited in Genbank (OR573691 and PP332812 respectively). The homology of 16S rDNA resulted over 98% with B. glumae (OK559611 and ON870618.1) and 100% with B. glumae (PP332812 and KX213523) for gyrB gene. To confirm B. glumae by pathogenicity test, 10 mL (108 UFC/ml) suspension of the representative strains, 0.5 mL was then injected into the panicles and sheaths of Horidhan (a susceptible local variety) in greenhouse condition and a control was inoculated with distilled water (Nandakumar et al. 2009). Typical BPB like symptoms were observed after 3 weeks post inoculation. The pathogen was again confirmed by reisolating from the infected spots as B. glumae to fulfill Koch's postulates. This report confirms the presence of B. glumae causing BPB of rice in Bangladesh. Future research for the investigation of BPB and the evolutionary origins of its causal bacteria is necessary to reduce the emergence of the disease and its management in Bangladesh.

Composition and Distribution of Bacteria, Pathogens, and Antibiotic Resistance Genes at Shanghai Port, China

As one of the biggest ports, the aquatic ecology of Shanghai Port has a significant impact on the health of surrounding residents and aquatic animals. Therefore, this study explored the composition and distribution of bacteria, pathogens, and ARGs at Shanghai Ports by high-throughput sequencing. The results showed that the bacterial community was dominated by the Proteobacteria at the phylum level. With the refinement of classification level, the main composition of bacteria in different samples shows significant differences. In contrast, there was no significant difference in the composition and distribution of pathogens among the samples. It indicates that pathogens and ARGs have steady community composition at Shanghai Port. Pseudomonas aeruginosa and Salmonella enterica could be regarded as candidate indicator pathogens. macB, tetA (58), and evgS were the main ARGs, and they showed a close relationship with Mycobacterium tuberculosis, Magnaporthe oryzae, Streptococcus pneumonia, Xanthomonas oryzae, and Vibrio cholerae in different sites and layers. This may lead to the transfer of ARGs between different pathogens, resulting in more threatening pathogens. The results showed that the pathogens and ARGs present posed a risk of transmission and were potentially harmful to residents in the environment. They can spread to any other ports in the world through the transmission of ballast water, posing a threat to humans and local ecology. In order to keep human health, it is necessary to pay more attention to the water environment of ports and increase the monitoring. Furthermore, it is essential to strengthen the supervision of external source pollution input due to the special features of the port.

Advances in breeding, biotechnology, and nanotechnological approaches to combat sheath blight disease in rice.

Sheath blight, caused by the fungus Rhizoctonia solani, is a major problem that significantly impacts rice production and can lead to substantial yield losses. The disease has become increasingly problematic in recent years due to the widespread use of high-yielding semi-dwarf rice cultivars, dense planting, and heavy application of nitrogenous fertilizers. The disease has become more challenging to manage due to its diverse host range and the lack of resistant cultivars. Despite utilizing traditional methods, the problem persists without a satisfactory solution. Therefore, modern approaches, including advanced breeding, transgenic methods, genome editing using CRISPR/Cas9 technology, and nanotechnological interventions, are being explored to develop rice plants resistant to sheath blight disease. This review primarily focuses on these recent advancements in combating the sheath blight disease.

Multi-Population Analysis for Leaf and Neck Blast Reveals Novel Source of Neck Blast Resistance in Rice.

Rice blast is one of the most devastating biotic stresses that limits rice productivity. The North Eastern Hill (NEH) region of India is considered to be one of the primary centres of diversity for both rice and pathotypes of Magnaporthe grisea. Therefore, the present study was carried out to elucidate the genetic basis of leaf and neck blast resistance under Meghalaya conditions. A set of 80 diverse genotypes (natural population) and 2 F2 populations involving resistant parent, a wildtype landrace, LR 5 (Lal Jangali) and susceptible genotypes Sambha Mahsuri SUB 1 (SMS) and LR 26 (Chakhao Poireiton) were used for association analysis of reported major gene-linked markers with leaf and neck blast resistance to identify major effective genes under local conditions. Genotyping using twenty-five gene-specific markers across diverse genotypes and F2 progenies revealed genes Pi5 and Pi54 to be associated with leaf blast resistance in all three populations. Genes Pib and qPbm showed an association with neck blast resistance in both natural and LR 5 × SMS populations. Additionally, a set of 184 genome-wide polymorphic markers (SSRs and SNPs), when applied to F2-resistant and F2-susceptible DNA bulks derived from LR 5 × LR 26, suggested that Pi20(t) on chromosome 12 is one of the major genes imparting disease resistance. Markers snpOS318, RM1337 and RM7102 and RM247 and snpOS316 were associated with leaf blast and neck blast resistance, respectively. The genotypes, markers and genes will help in marker-assisted selection and development of varieties with durable resistance.

Rice transcription factor DPF regulates stress-induced biosynthesis of diterpenoid phytoalexins.

Diterpenoid Phytoalexin Factor (DPF) is a key transcription factor involved in diterpenoid phytoalexin (DP) biosynthesis under non-stressed conditions in rice (Oryza sativa L.). Using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9, DPF knockout rice lines were generated. Treatments with abiotic stresses (copper chloride, ultraviolet light, and jasmonic acid) and biotic stress (blast fungus infection) to the knockout lines revealed that the DPF positively regulates stress-induced DP biosynthesis.

The characterization, pathotype distribution and genetic diversity of Xanthomonas oryzae pv. oryzae in rice production centers of the Special Region of Yogyakarta, Indonesia

Xanthomonas oryzae pv. oryzae (Xoo) is a pathogenic bacterium that induces bacterial leaf blight in rice. This disease is challenging to treat due to the presence of multiple pathotypes that can harm the plants. Characterizing and determining the distribution of pathotypes and genetic diversity of Xoo in the Special Region of Yogyakarta is essential for evaluating the appropriate approach to managing rice leaf blight in different districts of Yogyakarta. This study aimed to ascertain the characterization, distribution of pathotypes, and genetic diversity of Xoo in the Special Region of Yogyakarta. This study involved the isolation and characterization of Xoo from various rice production centers in Yogyakarta. The distribution of Xoo pathotypes was determined using five differential varieties (Tetep (4251), PB 5 (4827), Java 14 (11022), Kencana Bali (4477), and Kuntulan (1529)). Pathogenicity testing was conducted on six common varieties used by Yogyakarta farmers (IR64, Ciherang, C4, Mekongga, Menthik Wangi, and Inpari 42). Additionally, molecular characterization of Xoo was performed. The bacterial leaf blight that affects rice plants in the Yogyakarta region, caused by Xoo, is identified by yellow circular colonies. It exhibits a negative Gram staining response, positive catalase activity, negative oxidase activity, and does not hydrolyse starch. The Xoo pathotypes identified in Yogyakarta are IV, VIII, and XI. All six prevalent cultivars utilized by farmers in Yogyakarta are susceptible to Xoo. Out of each pathotype, four isolates were chosen, and they were divided into two distinct groups based on the DNA banding pattern they formed. Among these isolates, three had the lowest base sequence at 200 bp, while one isolate had a different DNA banding pattern with the lowest base sequence between 250–300 bp.

The characterization, pathotype distribution and genetic diversity of Xanthomonas oryzae pv. oryzae in rice production centers of the Special Region of Yogyakarta, Indonesia

Xanthomonas oryzae pv. oryzae (Xoo) is a pathogenic bacterium that induces bacterial leaf blight in rice. This disease is challenging to treat due to the presence of multiple pathotypes that can harm the plants. Characterizing and determining the distribution of pathotypes and genetic diversity of Xoo in the Special Region of Yogyakarta is essential for evaluating the appropriate approach to managing rice leaf blight in different districts of Yogyakarta. This study aimed to ascertain the characterization, distribution of pathotypes, and genetic diversity of Xoo in the Special Region of Yogyakarta. This study involved the isolation and characterization of Xoo from various rice production centers in Yogyakarta. The distribution of Xoo pathotypes was determined using five differential varieties (Tetep (4251), PB 5 (4827), Java 14 (11022), Kencana Bali (4477), and Kuntulan (1529)). Pathogenicity testing was conducted on six common varieties used by Yogyakarta farmers (IR64, Ciherang, C4, Mekongga, Menthik Wangi, and Inpari 42). Additionally, molecular characterization of Xoo was performed. The bacterial leaf blight that affects rice plants in the Yogyakarta region, caused by Xoo, is identified by yellow circular colonies. It exhibits a negative Gram staining response, positive catalase activity, negative oxidase activity, and does not hydrolyse starch. The Xoo pathotypes identified in Yogyakarta are IV, VIII, and XI. All six prevalent cultivars utilized by farmers in Yogyakarta are susceptible to Xoo. Out of each pathotype, four isolates were chosen, and they were divided into two distinct groups based on the DNA banding pattern they formed. Among these isolates, three had the lowest base sequence at 200 bp, while one isolate had a different DNA banding pattern with the lowest base sequence between 250–300 bp.

Rmg10, a Novel Wheat Blast Resistance Gene Derived from Aegilops tauschii.

Wheat blast, caused by Pyricularia oryzae (syn. Magnaporthe oryzae) pathotype Triticum (MoT), is a devastating disease that can result in up to 100% yield loss in affected fields. To find new resistance genes against wheat blast, we screened 199 accessions of Aegilops tauschii, the D genome progenitor of common wheat (Triticum aestivum), by seedling inoculation assays with Brazilian MoT isolate Br48 and found 14 resistant accessions. A synthetic hexaploid wheat line (Ldn/KU-2097) derived from a cross between the T. turgidum 'Langdon' (Ldn) and resistant A. tauschii accession KU-2097 exhibited resistance in seedlings and spikes against Br48. In an F2 population derived from 'Chinese Spring' × Ldn/KU-2097, resistant and susceptible individuals segregated in a 3:1 ratio, suggesting that the resistance from KU-2097 is controlled by a single dominant gene. We designated this gene Rmg10. Genetic mapping using an F2:3 population from the same cross mapped the RMG10 locus to the short arm of chromosome 2D. Rmg10 was ineffective against Bangladesh isolates but effective against Brazilian isolates. Field tests in Bolivia showed increased spike resistance in a synthetic octaploid wheat line produced from a cross between common wheat cultivar 'Gladius' and KU-2097. These results suggest that Rmg10 would be beneficial in farmers' fields in South America.

Enhancing resistance to bacterial blight in rice using CRISPR-based base editing technology

Bacterial blight (BB), caused by Xanthomonas oryzae pathovar oryzae (Xoo), poses a significant threat to rice production, particularly in Asia and West Africa. Breeding resistance against BB in elite rice varieties is crucial to advancing rice breeding program and supporting smallholder farmers. Transcription Activator-Like effectors (TALes) are key virulence factors in Xoo, with some targeting the susceptibility (S) genes such as the sugar transporter SWEET genes in rice. Among these, SWEET14 is an important S gene, with its promoter bound by the TALe TalC which exists across all sequenced African Xoo isolates. In the present study, we utilized CRISPR/Cas9-based cytidine and adenine base editors to alter the effector binding element (EBE) of TalC in the promoter of SWEET14 in rice cultivars Kitaake, IR24, and Zhonghua 11. Mutations with C to T changes in EBE led to reduced SWEET14 induction by TalC-containing Xoo strains, resulting in resistance to African Xoo isolates reliant on TalC for virulence. Conversely, A to G changes retained SWEET14 inducibility and susceptibility to Xoo in edited lines. Importantly, no off-target mutations were detected at predicted sites, and the edited lines exhibited no obvious defects in major agronomic traits in Kitaake. These results underscore the effectiveness of base editing systems for both molecular biology research and crop improvement endeavors.

A distinct protein posttranslational modifications-linked OsATL32-OsPPKL2-OsGSK2 loop modulates rice immunity against blast disease.

Protein posttranslational modifications play crucial roles in plant immunity through modulating a complicated signaling network mediated by different hormones. We previously demonstrated that OsATL32, an ATL-type E3 ligase, negatively contributes to rice immunity against Magnaporthe oryzae. Here, we show that OsATL32 forms a loop with OsPPKL2 and OsGSK2 through distinct protein posttranslational modifications to modulate rice immunity. OsATL32 ubiquitinates OsPPKL2, a protein phosphatase with Kelch-like repeat domains that exerts positive roles in regulating rice immunity against M. oryzae and chitin-triggered immune responses, for degradation. The glycogen synthase kinase 2 (OsGSK2), which acts as a negative regulator of rice immunity against M. oryzae and chitin-triggered immune responses, phosphorylates OsATL32 to elevate its protein stability and E3 ligase activity on OsPPKL2. Moreover, OsPPKL2 directly dephosphorylates OsGSK2, affecting its kinase activity on substrates including OsATL32 for phosphorylation. Like OsGSK2 as a BR signaling repressor, OsATL32 negatively regulates BR signaling; conversely, OsPPKL2 plays a positive role in BR signaling. These findings provide a molecular mechanism in which OsATL32 serves as a node connecting BR signaling and immunity by associating with OsPPKL2 and OsGSK2, assembling into a distinct protein posttranslational modifications-linked loop that functions in rice BR signaling and immunity.

Molecular Evolution of Rice Blast Resistance Gene bsr-d1

Rice blast, caused by the fungus Magnaporthe oryzae, reduces rice yields by 10% to 35%. Incorporating plant blast resistance genes into breeding programs is an effective strategy to combat this disease. Understanding the genetic variants that confer resistance is crucial to this strategy. The gene Bsr-d1 encodes a C2H2-like transcription factor, and its recessive allele confers broad-spectrum resistance against infections by various strains of M. oryzae. In this study, we investigated the molecular evolution of the rice blast resistance gene bsr-d1 in a representative population consisting of 827 cultivated and wild rice varieties. Our results revealed that wild rice exhibited significantly higher nucleotide diversity, with polymorphic regions primarily concentrated in the promoter region, in contrast to indica and japonica rice varieties. The Bsr-d1 gene displayed significant differentiation between indica and japonica, with the bsr-d1 resistance allele being unique to indica. Haplotype network and phylogenetic analyses suggested that the bsr-d1 resistance allele most likely originated from Oryza nivara in the region adjacent to the Indian Peninsula and the Indochina Peninsula. Moreover, we explored the utilization of bsr-d1 resistance alleles in China and designed a pair of DNA primers based on the polymorphic sites for the detection of the bsr-d1 resistance gene. In summary, our study uncovering the origin and evolution of bsr-d1 will enhance our comprehension of resistance gene variation and expedite the resistance breeding process.

Influence of weather parameters on rice blast disease progression in Tamil Nadu, India

Rice cultivation in Madurai district, Tamil Nadu, spans distinct cropping seasons: Kar (May – Jun), Semi-dry (Jul – Aug), Samba/ Late Samba (Aug – Sep), and Navarai (Dec – Jan), each with unique weather conditions and rice varieties. This study explores the correlation between weather parameters temperature, rainfall, humidity, sunshine hours, and wind speed and rice blast disease severity from 2021 to 2023. Using multiple linear regression with ordinary least squares (OLS), the analysis achieves high predictive accuracy (R² = 0.98). Results show that the maximum temperatures correlated negatively with disease severity (r = -0.869 to -0.892), while rainfall (r = 0.768 to 0.804) and wind speed (r = 0.766 to 0.938) correlated positively during the semi-dry season. Relative humidity exhibits varying impacts across seasons. These findings underscore the importance of tailored disease management strategies, such as targeted fungicidal applications during warmer seasons and optimized water management in others. By elucidating these dynamics, the study enhances understanding of weather-disease interactions, providing actionable insights to optimize disease management and enhance crop resilience in Madurai district.

Fine-tuning of IPA1 transactivation activity by E3 ligase IPI7-mediated non-proteolytic K29-ubiquitination during Magnaporthe oryzae infection

The Ideal Plant Architecture 1 (IPA1) transcription factor promotes rice yield and immunity through phosphorylation at its amino acid residue Ser163 as a switch. Although phosphorylated IPA1 mimic, IPA1(S163D), directly targets the promoter of immune response gene WRKY45, it cannot activate its expression. Here, we identified a co-activator of IPA1(S163D), a RING-finger E3 ligase IPA1 interactor 7 (IPI7), which fine-tunes the transcriptional activity of IPA1 to timely promote plant immunity and simultaneously maintain growth for yield. IPI7 interacts with IPA1 and promotes K29-polyubiquitination of IPA1 in vitro and in vivo. However, the stability of IPA1 protein is not affected by IPI7-mediated ubiquitination. The IPI7-promoted K29-polyubiquitination of IPA1 is induced by Magnaporthe oryzae infection and required for phosphorylated IPA1 to transactivate WRKY45 expression for immune response but not for plain IPA1 to transactivate DENSE AND ERECT PANICLES 1 (DEP1) expression for panicle development. IPI7 knockout impairs IPA1-mediated immunity but not yield. Our study reveals that plants utilize non-proteolytic K29-ubiquitination as a response to pathogen infection to fine-tune IPA1 transactivation activity for promoting immunity.

Priming of Exogenous Salicylic Acid under Field Conditions Enhances Crop Yield through Resistance to Magnaporthe oryzae by Modulating Phytohormones and Antioxidant Enzymes.

This study explores the impact of exogenous salicylic acid (SA) alongside conventional treatment by farmers providing positive (Mancozeb 80 % WP) and negative (water) controls on rice plants (Oryza sativa L.), focusing on antioxidant enzyme activities, phytohormone levels, disease resistance, and yield components under greenhouse and field conditions. In greenhouse assays, SA application significantly enhanced the activities of peroxidase (POX), polyphenol oxidase (PPO), catalase (CAT), and superoxide dismutase (SOD) within 12-24 h post-inoculation (hpi) with Magnaporthe oryzae. Additionally, SA-treated plants showed higher levels of endogenous SA and indole-3-acetic acid (IAA) within 24 hpi compared to the controls. In terms of disease resistance, SA-treated plants exhibited a reduced severity of rice blast under greenhouse conditions, with a significant decrease in disease symptoms compared to negative control treatment. The field study was extended over three consecutive crop seasons during 2021-2023, further examining the efficacy of SA in regular agricultural practice settings. The SA treatment consistently led to a reduction in rice blast disease severity across all three seasons. Yield-related parameters such as plant height, the number of tillers and panicles per hill, grains per panicle, and 1000-grain weight all showed improvements under SA treatment compared to both positive and negative control treatments. Specifically, SA-treated plants yielded higher grain outputs in all three crop seasons, underscoring the potential of SA as a growth enhancer and as a protective agent against rice blast disease under both controlled and field conditions. These findings state the broad-spectrum benefits of SA application in rice cultivation, highlighting its role not only in bolstering plant defense mechanisms and growth under greenhouse conditions but also in enhancing yield and disease resistance in field settings across multiple crop cycles. This research presents valuable insights into the practical applications of SA in improving rice plant resilience and productivity, offering a promising approach for sustainable agriculture practices.

Detection of Rice Leaf SPAD and Blast Disease Using Integrated Aerial and Ground Multiscale Canopy Reflectance Spectroscopy

Rice blast disease is one of the major diseases affecting rice plant, significantly impacting both yield and quality. Current detecting methods for rice blast disease mainly rely on manual surveys in the field and laboratory tests, which are inefficient, inaccurate, and limited in scale. Spectral and imaging technologies in the visible and near-infrared (Vis/NIR) region have been widely investigated for crop disease detection. This work explored the potential of integrating canopy reflectance spectra acquired near the ground and aerial multispectral images captured with an unmanned aerial vehicle (UAV) for estimating Soil-Plant Analysis Development (SPAD) values and detecting rice leaf blast disease in the field. Canopy reflectance spectra were preprocessed, followed by effective band selection. Different vegetation indices (VIs) were calculated from multispectral images and selected for model establishment according to their correlation with SPAD values and disease severity. The full-wavelength canopy spectra (450–850 nm) were first used for establishing SPAD inversion and blast disease classification models, demonstrating the effectiveness of Vis/NIR spectroscopy for SPAD inversion and blast disease detection. Then, selected effective bands from the canopy spectra, UAV VIs, and the fusion of the two data sources were used for establishing corresponding models. The results showed that all SPAD inversion models and disease classification models established with the integrated data performed better than corresponding models established with the single of either of the aerial and ground data sources. For SPAD inversion models, the best model based on a single data source achieved a validation determination coefficient (Rcv2) of 0.5719 and a validation root mean square error (RMSECV) of 2.8794, while after ground and aerial data fusion, these two values improved to 0.6476 and 2.6207, respectively. For blast disease classification models, the best model based on a single data source achieved an overall test accuracy of 89.01% and a Kappa coefficient of 0.86, and after data fusion, the two values improved to 96.37% and 0.95, respectively. These results indicated the significant potential of integrating canopy reflectance spectra and UAV multispectral images for detecting rice diseases in large fields.

Research on Lightweight Rice False Smut Disease Identification Method Based on Improved YOLOv8n Model

In order to detect rice false smut quickly and accurately, a lightweight false smut detection model, YOLOv8n-MBS, was proposed in this study. The model introduces the C2f_MSEC module to replace C2f in the backbone network for better extraction of key features of false smut, enhances the feature fusion capability of the neck network for different sizes of false smut by using a weighted bidirectional feature pyramid network, and designs a group-normalized shared convolution lightweight detection head to reduce the number of parameters in the model head to achieve model lightweight. The experimental results show that YOLOv8n-MBS has an average accuracy of 93.9%, a parameter count of 1.4 M, and a model size of 3.3 MB. Compared with the SSD model, the average accuracy of the model in this study increased by 4%, the number of parameters decreased by 89.8%, and the model size decreased by 86.9%; compared with the YOLO series of YOLOv7-tiny, YOLOv5n, YOLOv5s, and YOLOv8n models, the YOLOv8n-MBS model showed outstanding performance in terms of model accuracy and model performance detection; compared to the latest YOLOv9t and YOLOv10n models, the average model accuracy increased by 2.8% and 2.2%, the number of model parameters decreased by 30% and 39.1%, and the model size decreased by 29.8% and 43.1%, respectively. This method enables more accurate and lighter-weight detection of false smut, which provides the basis for intelligent management of rice blast disease in the field and thus promotes food security.

Characterization of Siderophore Produced by Bacillus velezensis YL2021 and Its Application in Controlling Rice Sheath Blight and Rice Blast.

Bacillus velezensis YL2021 has extensive antimicrobial activities against phytopathogens, and its genome harbors a catechol-type siderophore biosynthesis gene cluster. Here, we describe the characterization of siderophore produced by strain YL2021 and its antimicrobial activity in vitro and in vivo. A few types of siderophores were detected by chrome azurol S plates coupled with Arnow's test, purified and identified by Reversed-phase high-performance liquid chromatography (RP-HPLC). We found that strain YL2021 can produce different antimicrobial compounds under low-iron M9 medium or iron-sufficient LB medium although antimicrobial activities can be easily observed on the two media as described above in vitro. Strain YL2021 can produce at least three catechol-type siderophores in low-iron M9 medium while no siderophore was produced in LB medium. Among them, the main antimicrobial siderophore produced by strain YL2021 was bacillibactin, with m/z of 882, based on the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, which has broad-spectrum antimicrobial activities against Gram-positive, Gram-negative bacteria, the oomycete Phytophthora capsici and phytopathogenic fungi. Moreover, the antifungal activity of siderophore including bacillibactin observed in vitro was correlated with control efficacies against rice sheath blight disease caused by Rhizoctonia solani and rice blast disease caused by Magnaporthe oryzae in vivo. Collectively, the results demonstrate that siderophore including bacillibactin produced by Bacillus velezensis YL2021 is a promising biocontrol agent for application in rice disease control.

Antagonistic Mechanism Analysis of Bacillus velezensis JLU-1, a Biocontrol Agent of Rice Pathogen Magnaporthe oryzae.

Magnaporthe oryzae, the causal agent of rice blast, is a fungal disease pathogen. Bacillus spp. have emerged as the most promising biological control agent alternative to chemical fungicides. In this study, the bacterial strain JLU-1 with significant antagonistic activity isolated from the rhizosphere soil of rice was identified as Bacillus velezensis through whole-genome sequencing, average nucleotide identity analysis, and 16S rRNA gene sequencing. Twelve gene clusters for secondary metabolite synthesis were identified in JLU-1. Furthermore, 3 secondary metabolites were identified in JLU-1, and the antagonistic effect of secondary metabolites against fungal pathogens was confirmed. Exposure to JLU-1 reduced the virulence of M. oryzae, and JLU-1 has the ability to induce the reactive oxygen species production of rice and improve the salt tolerance of rice. All of these results indicated that JLU-1 and its secondary metabolites have the promising potential to be developed into a biocontrol agent to control fungal diseases.

TRNA-m1A methylation controls the infection of Magnaporthe oryzae by supporting ergosterol biosynthesis

Ergosterols are essential components of fungal plasma membranes. Inhibitors targeting ergosterol biosynthesis (ERG) genes are critical for controlling fungal pathogens, including Magnaporthe oryzae, the fungus that causes rice blast. However, the translational mechanisms governing ERG gene expression remain largely unexplored. Here, we show that the Trm6/Trm61 complex catalyzes dynamic N1-methyladenosine at position 58 (m1A58) in 51 transfer RNAs (tRNAs) of M.oryzae, significantly influencing translation at both the initiation and elongation stages. Notably, tRNA m1A58 promotes elongation speed at most cognate codons mainly by enhancing eEF1-tRNA binding rather than affecting tRNA abundance or charging. The absence of m1A58 leads to substantial decreases in the translation of ERG genes, ergosterol production, and, consequently, fungal virulence. Simultaneously targeting the Trm6/Trm61 complex and the ergosterol biosynthesis pathway markedly improves rice blast control. Our findings demonstrate an important role of m1A58-mediated translational regulation in ergosterol production and fungal infection, offering a potential strategy for fungicide development.

A diverse Fusarium community is responsible for contamination of rice with a variety of Fusarium toxins

Rice plays an important role in the daily diet in China and therefore its quality and safety have been of great concern. However, few systematic studies have investigated Fusarium community and toxins in rice grains. Here, we collected 1381 rice samples from Jiangsu Province in eastern China and found a higher frequency of zearalenone (ZEN), deoxynivalenol (DON), fumonisins (FBs), and beauvericin (BEA). The positive samples were individually contaminated with a minimum of one and a maximum of ten toxins. Fusarium was isolated and identified as the major fungus, which exhibited temporal and geographical distribution. The most prevalent species complexes within this genus were Fusarium incarnatum-equiseti species complex (FIESC), Fusarium fujikuroi species complex (FFSC), and Fusarium sambucinum species complex (FSAMSC). Nevertheless, the amplicon sequence analysis revealed a low relative abundance of Fusarium in the rice panicles, and the fungal community exhibited an irregular change along with the symptom’s emergence. In vitro toxigenic profiles of Fusarium strains showed significant complexity and specificity depending on the type and content. FIESC strains were non-pathogenic to wheat heads and weakly pathogenic to maize ears, respectively, accumulating lower amounts of toxins than F. asiaticum and F. fujikuroi. There was no significant variation in the ability to cause panicle blight in rice among the various species tested. Our study provides detailed information about the contamination of Fusarium toxins and community in rice after harvest. This information is valuable for understanding the relationship between Fusarium and rice and for developing effective control strategies.