Data_Sheet_1_Role of seed infection for the near and far distance dissemination of wheat blast caused by Magnaporthe oryzae pathotype Triticum.pdf

Magnaporthe oryzae pathotype Triticum (MoT) is a devastating fungal phytopathogen causing wheat blast disease which threatens wheat production particularly in warmer climate zones. Effective disease control is hampered by the limited knowledge on the life cycle, epidemiology, and pathogenicity of MoT. Since MoT mainly infects and colonizes the inflorescences of wheat, infection, invasion routes and colonization of MoT on wheat ears and in wheat seeds were investigated in order to assess potential seed transmission pathways. MoT was spray inoculated on two wheat cultivars (Sumai 3, susceptible and Milan, resistant) at three ear maturity stages [full ear emergence, growth stage (GS) 59; mid flowering, GS 65; and end of flowering, GS 69]. Incidence of MoT on Sumai 3 seeds was 100% and 20–25% on Milan. MoT sporulation rate on Sumai 3 contaminated seeds was more than 15 times higher than on Milan. Repeated washes of seed samples for removing paraffin fixation hampers seed microscopy. To overcome the damage of seed samples, we used hand-sectioned seed samples instead of paraffin-fixed microtome samples to facilitate microscopy. The colonization of MoT within various seed tissues was followed by light and confocal laser scanning microscopy (CLSM). Invasion of MoT in seeds predominantly occurred in the caryopsis germ region, but entry via other seed parts was also observed, confirming the potential of intense colonization of MoT in wheat grains. Fungal spread in wheat plants growing from MoT infected seeds was monitored through plating, microscopic and molecular techniques. Under greenhouse conditions, no spread of MoT from infected seeds to seedlings later than GS 21 or to ears was detected, neither in Milan nor in Sumai 3. We therefore conclude, that MoT may not systemically contaminate inflorescences and seeds in neither susceptible nor resistant wheat cultivars. However, initial blast symptoms, only found on seedlings of Sumai 3 but not Milan, resulted in the formation of new conidia, which may serve as inoculum source for plant-to-plant dissemination by airborne infection of plant stands in the field (short distance spread). Ultimately the inoculum may infect young inflorescences in the field and contaminate seeds. Our findings again stress the risk of long-distance dissemination of wheat blast across continents through MoT-contaminated seeds. This underlines the importance of mandatory use of healthy seeds in strategies to control any further spread of wheat blast.</p

Transcriptome sequencing of infected wheat samples collected during the wheat blast epidemic in Bangladesh in 2017 and release of raw sequence data on OpenWheatBlast website with open access

Wheat fields in Bangladesh were again infected by wheat blast fungus (<i>Magnaporthe oryzae</i>) in 2017 growing season, following the epidemic of 2016. Infected wheat samples were collected by Tofazzal Islam and team, and sent to the Sainsbury Laboratory for RNA extraction and sequencing. Library preparation and RNA-Seq sequencing runs were performed on Illumina HiSeq-2500 machines at the Earlham Institute to produce paired-end reads with ~267 bp average insert size. Here we report the release of these data to general public with open access on OpenWheatBlast website.<br

Transcriptome sequencing of infected wheat samples collected during the wheat blast epidemic in Bangladesh in 2016 and release of raw sequence data on OpenWheatBlast website for open access

In 2016, wheat fields in Bangladesh were infected by wheat blast fungus (<i>Magnaporthe oryzae</i>) for the first time. The fungus spread quickly thoroughout the country and threatened other wheat growing areas in neighbouring countries. Islam <i>et al </i>(BMC Biology 2016 14:84<strong>, </strong>DOI<strong>: </strong>10.1186/s12915-016-0309-7) collected infected wheat samples, extracted RNA, and obtained short reads by Illumina RNA-Seq technology. Library preparation and sequencing runs were performed on Illumina HiSeq-2500 machines to produce paired-end reads with ~300 bp insert size. Here we report the release of all raw sequence data to public via OpenWheatBlast website for open access.<br><div><br></div

The urgency of wider adoption of one health approach for the prevention of a future pandemic

Recurring outbreaks of emerging and re-emerging zoonoses serve as a reminder that the health of humans, animals, and the environment are interconnected. Therefore, multisectoral, transdisciplinary, and collaborative approaches are required at local, regional, and global levels to tackle the ever-increasing zoonotic threat. The ongoing pandemic of COVID-19 zoonosis has been posing tremendous threats to global human health and economies. The devastation caused by the COVID-19 pandemic teaches us to adopt a “One Health Approach (OHA)” to tackle a possible future pandemic through a concerted effort of the global scientific community, human health professionals, public health experts, veterinarians and policymakers through open science and open data sharing practices. The OHA is an integrated, holistic, collaborative, multisectoral, and transdisciplinary approach to tackle potential pandemic zoonotic diseases. It includes expanding scientific inquiry into zoonotic infections; monitoring, and regulating traditional food markets, transforming existing food systems, and incentivizing animal husbandry and legal wildlife trade to adopt effective zoonotic control measures. To adopt an OHA globally, research and academic institutions, governments and non-government sectors at the local, regional, and international levels must work together. This review aimed to provide an overview of the major pandemics in human history including the COVID-19, anthropogenic drivers of zoonoses, Severe Acute Respiratory Syndrome Coronavirus-2 (SARSCoV- 2) reverse zoonoses, the concept of OHA and how an OHA could be utilized to prevent future pandemic threats to the human-animal-ecosystem interfaces. In addition, this review article discusses the strategic framework of OHA and possible challenges to implement OHA in practice to prevent any future pandemics. The practices of open data sharing, open science, and international collaboration should be included in the OHA to prevent and/or rapidly tackle any health emergencies in the future

Key functions of target proteins.

Rice blast disease, caused by the fungus Magnaporthe oryzae, poses a severe threat to rice production, particularly in Asia where rice is a staple food. Concerns over fungicide resistance and environmental impact have sparked interest in exploring natural fungicides as potential alternatives. This study aimed to identify highly potent natural fungicides against M. oryzae to combat rice blast disease, using advanced molecular dynamics techniques. Four key proteins (CATALASE PEROXIDASES 2, HYBRID PKS-NRPS SYNTHETASE TAS1, MANGANESE LIPOXYGENASE, and PRE-MRNA-SPLICING FACTOR CEF1) involved in M. oryzae’s infection process were identified. A list of 30 plant metabolites with documented antifungal properties was compiled for evaluation as potential fungicides. Molecular docking studies revealed that 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin exhibited superior binding affinities compared to reference fungicides (Azoxystrobin and Tricyclazole). High throughput molecular dynamics simulations were performed, analyzing parameters like RMSD, RMSF, Rg, SASA, hydrogen bonds, contact analysis, Gibbs free energy, and cluster analysis. The results revealed stable interactions between the selected metabolites and the target proteins, involving important hydrogen bonds and contacts. The SwissADME server analysis indicated that the metabolites possess fungicide properties, making them effective and safe fungicides with low toxicity to the environment and living beings. Additionally, bioactivity assays confirmed their biological activity as nuclear receptor ligands and enzyme inhibitors. Overall, this study offers valuable insights into potential natural fungicides for combating rice blast disease, with 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin standing out as promising and environmentally friendly alternatives to conventional fungicides. These findings have significant implications for developing crop protection strategies and enhancing global food security, particularly in rice-dependent regions.</div

Contact frequency analysis for ML complexes.

A) ML-azoxystrobin complex and B) ML-rosmarinic acid. (DOCX)</p

Fig 4 -

(A) RMSF in side chain atoms of residues in CATALASE PEROXIDASES 2, HYBRID PKS-NRPS SYNTHETASE TAS1, MANGANESE LIPOXYGENASE, and PMSF proteins, and (B) Radius of gyration for corresponding systems of CATALASE PEROXIDASES 2, HYBRID PKS-NRPS SYNTHETASE TAS1, MANGANESE LIPOXYGENASE, and PMSF.</p

Fig 12 -

Cluster analysis for MANGANESE LIPOXYGENASE complexes: (A) MANGANESE LIPOXYGENASE-azoxystrobin complex and (B) MANGANESE LIPOXYGENASE-rosmarinic acid complex. Cluster analysis for PMSF complexes: (C) PMSF-azoxystrobin complex and (D) PMSF-myricetin complex.</p

Enlistment of antifungal plant metabolites.

Rice blast disease, caused by the fungus Magnaporthe oryzae, poses a severe threat to rice production, particularly in Asia where rice is a staple food. Concerns over fungicide resistance and environmental impact have sparked interest in exploring natural fungicides as potential alternatives. This study aimed to identify highly potent natural fungicides against M. oryzae to combat rice blast disease, using advanced molecular dynamics techniques. Four key proteins (CATALASE PEROXIDASES 2, HYBRID PKS-NRPS SYNTHETASE TAS1, MANGANESE LIPOXYGENASE, and PRE-MRNA-SPLICING FACTOR CEF1) involved in M. oryzae’s infection process were identified. A list of 30 plant metabolites with documented antifungal properties was compiled for evaluation as potential fungicides. Molecular docking studies revealed that 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin exhibited superior binding affinities compared to reference fungicides (Azoxystrobin and Tricyclazole). High throughput molecular dynamics simulations were performed, analyzing parameters like RMSD, RMSF, Rg, SASA, hydrogen bonds, contact analysis, Gibbs free energy, and cluster analysis. The results revealed stable interactions between the selected metabolites and the target proteins, involving important hydrogen bonds and contacts. The SwissADME server analysis indicated that the metabolites possess fungicide properties, making them effective and safe fungicides with low toxicity to the environment and living beings. Additionally, bioactivity assays confirmed their biological activity as nuclear receptor ligands and enzyme inhibitors. Overall, this study offers valuable insights into potential natural fungicides for combating rice blast disease, with 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin standing out as promising and environmentally friendly alternatives to conventional fungicides. These findings have significant implications for developing crop protection strategies and enhancing global food security, particularly in rice-dependent regions.</div

SwissADME properties of top metabolites.

Rice blast disease, caused by the fungus Magnaporthe oryzae, poses a severe threat to rice production, particularly in Asia where rice is a staple food. Concerns over fungicide resistance and environmental impact have sparked interest in exploring natural fungicides as potential alternatives. This study aimed to identify highly potent natural fungicides against M. oryzae to combat rice blast disease, using advanced molecular dynamics techniques. Four key proteins (CATALASE PEROXIDASES 2, HYBRID PKS-NRPS SYNTHETASE TAS1, MANGANESE LIPOXYGENASE, and PRE-MRNA-SPLICING FACTOR CEF1) involved in M. oryzae’s infection process were identified. A list of 30 plant metabolites with documented antifungal properties was compiled for evaluation as potential fungicides. Molecular docking studies revealed that 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin exhibited superior binding affinities compared to reference fungicides (Azoxystrobin and Tricyclazole). High throughput molecular dynamics simulations were performed, analyzing parameters like RMSD, RMSF, Rg, SASA, hydrogen bonds, contact analysis, Gibbs free energy, and cluster analysis. The results revealed stable interactions between the selected metabolites and the target proteins, involving important hydrogen bonds and contacts. The SwissADME server analysis indicated that the metabolites possess fungicide properties, making them effective and safe fungicides with low toxicity to the environment and living beings. Additionally, bioactivity assays confirmed their biological activity as nuclear receptor ligands and enzyme inhibitors. Overall, this study offers valuable insights into potential natural fungicides for combating rice blast disease, with 2-Coumaroylquinic acid, Myricetin, Rosmarinic Acid, and Quercetin standing out as promising and environmentally friendly alternatives to conventional fungicides. These findings have significant implications for developing crop protection strategies and enhancing global food security, particularly in rice-dependent regions.</div