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The fungal pathogen Magnaporthe oryzae causes devastating blast disease in various cereals, including rice (Oryza sativa), wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare). Despite previous reports on fungal host specificity, the mechanisms underlying differential host infection strategies remain unclear. This study aimed to identify differentially abundant proteins (DAPs) in the apoplast of rice, barley, and wheat following infection with two M. oryzae pathovars using liquid chromatography-tandem mass spectrometry (LC–MS/MS). LC–MS/MS analysis revealed an enrichment of both M. oryzae and host proteins in the apoplast during the compatible reaction compared to the incompatible reaction. DAPs from M. oryzae involved in the host interaction included secreted extracellular enzymes (e.g., hydrolases), which were significantly increased in the M. oryzae Oryzae (MoO)-infected rice apoplast. Among host proteins, the proportion of protein-modifying enzymes increased in the M. oryzae Triticum (MoT)-infected rice and MoO-infected wheat apoplastic fluids, particularly rice glycosidases, peroxidases, and serine proteases, as well as wheat serine proteases. Furthermore, DAPs from MoL-infected rice were enriched in carbohydrate metabolism, suggesting that carbohydrate metabolism-related proteins may play a vital role in rice resistance to MoL. Additionally, protein-modifying and cytoskeletal proteins, as well as stress-responsive proteins, were enriched in the MoO-infected wheat apoplastic fluid. Finally, DAPs from both MoO- and MoL-infected barley were enriched in hydrogen peroxide catabolism, suggesting that peroxidases may be vital for barley resistance to M. oryzae. The identification of DAPs from both M. oryzae strains and the three host plants offers valuable insights into the host specificity mechanisms of M. oryzae in cereal crops. [ABSTRACT FROM AUTHOR]

Wheat blast caused by Pyricularia oryzae pathotype Triticum (MoT) has been transmitted from South America to Bangladesh and Zambia and is now spreading in these countries. To prepare against its further spread to Asian countries, we introduced Rmg8, a gene for resistance to wheat blast, into a Japanese elite cultivar, Chikugoizumi (ChI), through recurrent backcrosses and established ChI near-isogenic lines, #2-1-10 with the Rmg8/Rmg8 genotype and #4-2-10 with the rmg8/rmg8 genotype. A molecular analysis suggested that at least 96.6% of the #2-1-10 genome was derived from the recurrent parent ChI. The #2-1-10 line was resistant to MoT not only in primary leaves at the seedling stage but also in spikes and flag leaves at the heading stage. The strength of the resistance in spikes of this Rmg8 carrier was comparable to that of a carrier of the 2NS segment, which has been the only genetic resource released to farmers' fields for wheat blast resistance. On the other hand, the 2NS resistance was not expressed on leaves at the seedling stage nor flag leaves at the heading stage. Considering that leaf blast has been increasingly reported and regarded as an important inoculum source for spike blast, Rmg8 expressed at both the seedling and heading stages, or more strictly in both leaves and spikes, is suggested to be useful to prevent the spread of MoT in Asia and Africa. [ABSTRACT FROM AUTHOR]

Abstract The fungal pathogen Magnaporthe oryzae causes devastating blast disease in various cereals, including rice (Oryza sativa), wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare). Despite previous reports on fungal host specificity, the mechanisms underlying differential host infection strategies remain unclear. This study aimed to identify differentially abundant proteins (DAPs) in the apoplast of rice, barley, and wheat following infection with two M. oryzae pathovars using liquid chromatography-tandem mass spectrometry (LC–MS/MS). LC–MS/MS analysis revealed an enrichment of both M. oryzae and host proteins in the apoplast during the compatible reaction compared to the incompatible reaction. DAPs from M. oryzae involved in the host interaction included secreted extracellular enzymes (e.g., hydrolases), which were significantly increased in the M. oryzae Oryzae (MoO)-infected rice apoplast. Among host proteins, the proportion of protein-modifying enzymes increased in the M. oryzae Triticum (MoT)-infected rice and MoO-infected wheat apoplastic fluids, particularly rice glycosidases, peroxidases, and serine proteases, as well as wheat serine proteases. Furthermore, DAPs from MoL-infected rice were enriched in carbohydrate metabolism, suggesting that carbohydrate metabolism-related proteins may play a vital role in rice resistance to MoL. Additionally, protein-modifying and cytoskeletal proteins, as well as stress-responsive proteins, were enriched in the MoO-infected wheat apoplastic fluid. Finally, DAPs from both MoO- and MoL-infected barley were enriched in hydrogen peroxide catabolism, suggesting that peroxidases may be vital for barley resistance to M. oryzae. The identification of DAPs from both M. oryzae strains and the three host plants offers valuable insights into the host specificity mechanisms of M. oryzae in cereal crops.

Wheat blast caused by Pyricularia oryme pathotype Triticum has spread to Asia (Bangladesh) and Africa (Zambia) from the endemic region of South America. Wheat varieties with durable resistance are needed, but very limited resistance resources are currently available. After screening tetraploid wheat accessions, we found an exceptional accession St19 Triticum dicoccum, KU - 114), Primary leaves of St 19 were resistant not only to Brazilian isolate Br48 (a carrier of Type el of AVR-Rm#8) but also to Br48AAS, an AVR-Rmg8 disruptant of Br48, even at 30°C, suggesting that the resistance of St 19 is tolerant to high temperature and controlled by gene or genes other than Ring8. When an h POpulation derived from a cross between St 19 and St30 (a susceptible accession of T paleocolchicum KU - 191) was inoculated with Br48, resistant and susceptible seedlings segregated in a 3: 1 ratio, indicating that resistance of Stl 9 is conferred by a single gene, We designated this gene Ring 11. Molecular mapping revealed that the RMG] 1 locus is located on the short arm of chi-omosome 7A. Rmg 1 1 is effective not only against other two Brazilian isolates (Br5 and Brl 16.5) but also against Bangladeshi isolates (T-108 and T-109) at the seedling stage. At the heading stage. lines containing Ring 11 were highly susceptible to the Bangladeshi isolates but moderately resistant to the Brazilian isolates. Stacking of Ring / 1 withRmg8 and the 2NS segment is highly recommended to achieve durable wheat blast resistance. [ABSTRACT FROM AUTHOR]

Wheat blast, caused by Pyricularia oryzae Triticum lineage (PoTl), can infect wheat leaves and heads. More information is needed on the pathogen biology and disease epidemiology of the isolate PoTl for the integrated management of wheat blast in Brazilian wheat fields. This study aimed to characterize the incubation period, latent period, and the temporal progress of wheat blast and to fit nonlinear models, describing the nature of an epidemic of the PoTl isolate 12.1.146 compared with the PoTl isolate Py6038, throughout five successive infection cycles of PoTl on wheat leaves and heads. Wheat blast occurred in all infection cycles. The incubation period and latent period of the PoTl isolate 12.1.146 were significantly shorter than that of the PoTl isolate Py6038. The secondary inocula produced by the PoTl isolate on symptomatic wheat leaves caused blast symptoms when inoculated on wheat heads. The area under the disease progress curve (AUDPC) was calculated based on disease severity. In all infection cycles, the AUDPC of the PoTl isolate 12.1.146 was significantly higher than that of the PoTl isolate Py6038. Between the 1st and 5th infection cycle, a significant reduction was observed in the AUDPC of the PoTl isolates. The nonlinear logistic model describe the disease progress curves (DPCs) of PoTl isolates on wheat leaves and heads best. Our findings indicate that the PoTl isolates did not maintain the same temporal dynamics after five successive infection cycles. These findings may have implications for the integrated management of the wheat blast pathogen in Brazilian fields. [ABSTRACT FROM AUTHOR]

Bangladesh experienced widespread wheat blast infections for the first time in 2016. The outbreak of the disease has significantly affected wheat acreage and production. This study uses an 'action theory' framework to identify the determinants that affected wheat growers to adopt certain production practices to deal with the situation. We followed a multistage sampling procedure and interviewed 150 wheat growers from two severely blast-affected districts, Meherpur and Kustia. According to 91.3% of farmers, the most remarkable adoption strategies were 'improved intercultural practices,' 'shifting variety,' 'shifting crops', and 'undertaking off-farm activities,' of which farmers mostly followed the former. We also used multivariate probit model analysis to identify factors that shape farmers' adaptation choices in wheat blast-affected areas. The adaptation strategies are influenced mainly by farmers' education, primary occupation, family size, government incentives, extension services, access to Information and Communications Technology (ICT), and annual income. According to the notable similarities between the four adoption strategies, an adoption study should consider all potential factors influencing farmers' adoption decisions. Policy implications include strengthening extension services, developing tailored adaptation strategies, and conducting relevant research. [ABSTRACT FROM AUTHOR]

Summary: The fungal pathogen, Magnaporthe oryzae Triticum pathotype, causing wheat blast disease was first identified in South America and recently spread across continents to South Asia and Africa. Here, we studied the genetic relationship among isolates found on the three continents.Magnaporthe oryzae strains closely related to a South American field isolate B71 were found to have caused the wheat blast outbreaks in South Asia and Africa. Genomic variation among isolates from the three continents was examined using an improved B71 reference genome and whole‐genome sequences.We found strong evidence to support that the outbreaks in Bangladesh and Zambia were caused by the introductions of genetically separated isolates, although they were all close to B71 and, therefore, collectively referred to as the B71 branch. In addition, B71 branch strains carried at least one supernumerary mini‐chromosome. Genome assembly of a Zambian strain revealed that its mini‐chromosome was similar to the B71 mini‐chromosome but with a high level of structural variation.Our findings show that while core genomes of the multiple introductions are highly similar, the mini‐chromosomes have undergone marked diversification. The maintenance of the mini‐chromosome and rapid genomic changes suggest the mini‐chromosomes may serve important virulence or niche adaptation roles under diverse environmental conditions. [ABSTRACT FROM AUTHOR]

Wheat blast, caused by Pyricularia oryzae Triticum (PoT), is an emerging threat to global wheat production. The current understanding of the population biology of the pathogen and epidemiology of the disease has been based on phylogenomic studies that compared the wheat blast pathogen with isolates collected from grasses that were invasive to Brazilian wheat fields. In this study, we performed a comprehensive sampling of blast lesions in wheat crops and endemic grasses found in and away from wheat fields in Minas Gerais. A total of 1,368 diseased samples were collected (976 leaves of wheat and grasses and 392 wheat heads), which yielded a working collection of 564 Pyricularia isolates. We show that, contrary to earlier implications, PoT was rarely found on endemic grasses, and, conversely, members of grass-adapted lineages were rarely found on wheat. Instead, most lineages were host-specialized, with constituent isolates usually grouping according to their host of origin. With regard to the dominant role proposed for signalgrass in wheat blast epidemiology, we found only one PoT member in 67 isolates collected from signalgrass grown away from wheat fields and only three members of UrocMoa-adapted lineages among hundreds of isolates from wheat. Cross-inoculation assays on wheat and a signalgrass used in pastures (U. brizantha') suggested that the limited cross-infection observed in the field may be due to innate compatibility differences. Whether or not the observed level of crossinfection would be sufficient to provide an inoculum reservoir, or serve as a bridge between wheat growing regions, is questionable and, therefore, deserves further investigation. [ABSTRACT FROM AUTHOR]

Pyricularia oryzae, a blast fungus of gramineous plants, is composed of various host genus–specific pathotypes. The avirulence of an Avena isolate on wheat is conditioned by PWT3 and PWT4. We isolated the third avirulence gene from the Avena isolate and designated it as PWT7. PWT7 was effective as an avirulence gene only at the seedling stage or on leaves. PWT7 homologs were widely distributed in a subpopulation of the Eleusine pathotype and the Lolium pathotype but completely absent in the Triticum pathotype (the wheat blast fungus). The PWT7 homolog found in the Eleusine pathotype was one of the five genes involved in its avirulence on wheat. A comparative analysis of distribution of PWT7 and the other two genes previously identified in the Eleusine pathotype suggested that, in the course of parasitic specialization toward the wheat blast fungus, a common ancestor of the Eleusine, Lolium, Avena, and Triticum pathotypes first lost PWT6, secondly PWT7, and, finally, the function of PWT3. PWT7 or its homologs were located on core chromosomes in Setaria and Eleusine isolates but on supernumerary chromosomes in Lolium and Avena isolates. This is an example of interchromosomal translocations of effector genes between core and supernumerary chromosomes. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract The objective of this work was to evaluate the effect of the presence of 2NVS translocation on the inheritance of wheat resistance to head blast, in F2 segregating lines from two biparental crosses. The IPF 86766 and 'Santa Fe' genotypes were used as female parents (both 2NVS carriers), and 'BRS 404', as the common male parent (non 2NVS carrier). Using a mixture of three isolates of Pyricularia oryzae Triticum, wheat spikes and rachises were evaluated for bleaching severity at five (%BS_5dpi) and seven days post-inoculation (%BS_7dpi), as well as for the number of infection points (IPR) and pathogen sporulation (SPO). Chi-square analyses showed that segregation follows a 3:1 Mendelian hypothesis, with a major locus with a dominant allele: %BS_7dpi for IPF 86766 × 'BRS 404' and IPR for 'Santa Fe' x 'BRS 404'. Therefore, this is the first known study to show that the 2NVS translocation responsible for resistance to head blast is a major locus with a dominant allele. In a controlled environment, 'BRS 404' shows bleaching severity and SPO similar with those occurring on IPF 86766 and 'Santa Fe'.

Given the scarcity of information about inoculum sources of Pyricularia oryzae pathotype Triticum (PoT) under field conditions, this study quantified the conidia of PoT on wheat leaves during the crop cycle and linked them with the presence of blast in the spikes. The study was conducted involving two different production environments: nine commercial wheat fields and an experimental plot under sprinkler irrigation. Throughout the crop season, leaves were collected monthly and processed to quantify the number of conidia present in each gram of dry leaves. At physiological maturity of the crop, the fields were sampled to quantify the number of infected spikes/m2. Significant differences in the quantity and persistence of the pathogen during the cycle were seen under both environments. A strong positive correlation between the number of conidia present on the leaves during the crop growth cycle and the percentage of damaged spikes was established. [ABSTRACT FROM AUTHOR]

Magnaporthe oryzae, the causal agent of blast diseases, is a destructive filamentous fungus that infects many plants including most economically important food crops, rice, wheat, pearl millet and finger millet. Magnaporthe oryzae has numerous pathotypes because of its high host-specificity in the field. The Oryza pathotype (MoO) of M. oryzae is the most devastating pathogen of rice, causing 10–30% yield loss in the world. On the other hand, the Triticum pathotype (MoT) causes blast disease in wheat, which is now a serious threat to wheat production in some South American countries, Bangladesh and Zambia. Because of low fungicide efficacy against the blast diseases and lack of availability of resistant varieties, control of rice and wheat blast diseases is difficult. Therefore, an integrated management programme should be adopted to control these two diseases in the field. Here, we introduced and summarized the classification, geographical distribution, host range, disease symptoms, biology and ecology, economic impact, and integrated pest management (IPM) programme of both rice and wheat blast diseases.

In this study Pyricularia spp., P. oryzae and the P. oryzae pathotype Triticum (PoT) were detected and identified in leaf segments of forage and invasive grasses located in or next to wheat fields. In 2018 and 2019, 66 samples of lesion leaf segments of Urochloa and other grasses were collected in Londrina (PR), Patos de Minas (MG), and Uberaba (MG). The detection and/or identification of the pathogens on the samples was conducted using moist chamber procedures and with the primers MoT3 and Pot2 by PCR. There were DNA amplification with the primer MoT3 (specific for PoT) for 13 (19.69%) of the samples, all of them from Urochloa. The finding that Urochloa hosts PoT at a relatively high rate raises concerns about the importance which these plants may have on the wheat blast cycle as an alternative host for the pathogen and/or source of inoculum for the disease. [ABSTRACT FROM AUTHOR]

The emergence and spread of wheat blast caused by fungal pathogen Magnaporthe oryzae pathotype Triticum is a threat to global wheat production. The resistance level and genetic loci for blast resistance in Chinese germplasm remain unknown. A panel of 266 bread wheat accessions from China, CIMMYT-Mexico and other countries was screened for head blast resistance under 12 field experiments in Bolivia and Bangladesh. Subsequently, a genome-wide association study was performed to understand the genetic basis of wheat blast resistance. The average blast index of all the accessions was 53.7% ± 12.7%, and 10 accessions including Chinese accessions Yumai 10 and Yu 02321 showed moderate to high levels of blast resistance, accounting for only 3.8% in the panel. Fifty-eight significant SNPs clustered in a 28.9 Mb interval on the 2AS/2NS translocation region, explaining phenotypic variation between 10.0% and 35.0%. The frequency of the 2AS/2NS translocation in the Chinese accessions was as low as 4.5%. These results indicated that the 2NS fragment was the only major locus conferring resistance to wheat blast in this panel, and the resistant and moderately resistant lines identified could be deployed in breeding.

The present research work was conducted to evaluate the efficacy of eight fresh botanicals namely Azadirachta indica (Neem leaf), Allium cepa (Onion bulb), Allium sativum (Garlic), Curcuma longa (Turmeric), Zingiber officinale (Ginger), Allamanda cathartica (Allamanda leaf), Nigella sativa (Black cumin) and Aloe vera (Aloe vera) against Magnaporthe oryzae triticum (MoT) and solvent, ethanol (95%) was used for the phytochemical extraction of various plant parts. Three concentrations namely 1:1 (w/v), 1:0.50 (w/v) and 1:0.25 (w/v) of plant parts and ethanol were used for botanical extraction. The antifungal activity of botanicals against a virulent MoT isolate CHMoT07 was evaluated in-vitro using poison food technique. The lowest mycelia growth was recorded with Aloe vera (Aloe vera leaf) extracts and Nigella sativa (Black cumin seeds) extracts @ 1:1 w/v and @ 1:0.25 w/v with growth rate of 3.00 mm and 3.33 mm respectively at 7 days after inoculation, whereas the highest mycelia growth rate of MoT isolates was recorded in control plates both at 7 DAI and 14 DAI under in-vitro condition. [ABSTRACT FROM AUTHOR]

Background: Quinone outside inhibitor (QoI) fungicides have not been effective in controlling the wheat blast disease [Pyricularia oryzae Triticum lineage (PoTl)] in Brazil. The first report of resistance of PoTl to QoIs in this country occurred in 2015. This study aimed to test hypotheses about the changes in fitness parameters and competitive advantage of the QoI‐resistant (R) PoTl isolate group compared to the sensitive (S) isolate group. Mycelial growth on PDA medium and in vivo conidial production, incubation period and disease severity were analyzed as fitness parameters. The competitive ability was measured on wheat leaves and heads inoculated with mixtures of R:S isolates at the following proportions: 0S:100R, 20S:80R, 50S:50R, 80S:20R, 100S:0R, and 0S:0R. Results: The QoI‐R isolate group had significantly higher fitness than the sensitive isolate group, considering both in vitro and in vivo parameters. The highest in vivo conidial production on wheat leaves and the highest leaf and head disease severity were detected when resistant strains were predominant in the isolate's mixtures (20S:80R or 0S:100R proportions), in the absence of fungicide pressure. Conidia harvested from wheat blast lesions on leaves inoculated with 20S:80R and 0S:100R mixtures were resistant to QoIs in vitro assays based on discriminatory doses of the fungicide. Conclusion: Therefore, QoI resistance facilitated a higher fitness and a competitive advantage in PoTl, which contrasts with the evolutionary theory that associates a fitness cost to fungicide resistance. We discuss the evolutionary and ecological implications of the higher fitness as found in the fungicide‐resistant adapted populations of the wheat blast pathogen. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Wheat blast caused by Magnaporthe oryzae Pathotype triticum (MoT) becomes one of the common nuisances to the expansion of wheat production in Bangladesh That's why to find out a suitable control measure, fourteen chemicals and five Trichoderma isolates were evaluated against MoT in in-vitro condition during the cropping seasons of 2019-2020. In in-vitro conditions the efficacies of fungicides and biological agents i.e. Trichoderma isolates were evaluated by the measurement of percent inhibition of radial mycelial growth of MoT. The highest percentage of radial mycelial growth inhibition (100%) of MoT was obtained by Mancozeb 63% + Carbendazim 12% (Companion), Carbendazim (Autostin 50 WDG), Pyrochlostrobin 5% + Metiram 55% (Cabrio Top), Tebuconazol (Folicur EW 250) and Hexaconazol (Contaf 5EC) fungicides at low concentration (0.0125%) compared to other fungicides at 10 days after inoculation (DAI). Out of five Trichoderma asperellum isolates used in this study, the maximum percent of radial mycelial growth inhibition (58.24%) of MoT was achieved by Trichoderma isolate PMILTE-N51 (58.24%) followed by isolate PMILTE- N41 (53.85%) in the in-vitro dual culture assay at 7 DAI. In case of pot experiment, seven fungicides (Nativo 75WG, Companion, Autostin, Indofil M45, Supreme 250EW, Contaf, Amiscore 32.5 SC) at 0.1% & 0.3% and bioagents (Trichoderma asperellum, PMILTE-N51-suspension, PMILTE-N51- formulation & Trichoderma herzianum based IPM LAB- Formulation) were used to evaluate their efficacy in controlling wheat blast disease and their effects on different yield contributing parameters of wheat. Minimum percentage of blast incidence and severity were recorded for Supreme 250 EW @ 0.3% (14.88% and 2.21%) treated plants followed by PMILTE-N51 suspension compared to control (85.06% and 90.2%). In case of different yield contributing parameters viz. maximum number of ear/pot (13.67), number of total and healthy spikelets/ear (55.33 and 52.33), number of total and healthy grains/ear (50.33 and 45.67 ) were achieved by Amiscore 32.5 SC @ 0.3% followed by Supreme 250EW @ 0.3% and a maximum weight of 1000 grains/pot (55 gm) was recorded for PMILTE-N51-suspension, whereas controlled plants showed the lowest value for all the yield contributing parameters among the treatments used in this experiment. These results indicate that Amiscore 32.5 SC and Supreme 250 EW (@ 0.3%) showed the best results as chemical control approaches, where Trichoderma (PMILTE-N51-suspension) can be a better non-chemical management approach against wheat blast in Bangladesh. [Fundam Appl Agric 2022; 7(1.000): 47-59]

Wheat blast (Magnaporthe oryzae Triticum) in Bangladesh and South America is recognized as one major limiting factor of wheat production. Its control using chemical pesticides raises concerns about food safety and pesticide resistance, which have dictated the need for alternative blast management approach, nutrient supplementation could be an ecofriendly alternative. Experiments were carried out under confined net house condition for two consecutive cropping seasons. Single doses of the nutrients (Si, B and Zn) were incorporated during soil preparation. Plants of the wheat blast susceptible variety BARI Gom-26 were inoculated with spores (1 x 107 spores ml-1) of Magnaporthe oryzae Triticum at blast vulnerable pre-heading stage of 52 days age. Typical wheat blast symptoms of spike bleaching from top to downward appeared on sight 14 days after inoculation i.e., 66 days age of the crop. Incidence and severity of blast bleaching of spike were scored for four times starting from 68 days age @ three day’s interval. None of the nutrients could stop the incidence of blast on wheat; however, some nutrients reduced the blast incidence significantly. Solo application of Si, B and Zn or combination of two caused significant reduction of spike bleaching. With the mixed application of Si, B and Zn, > 47% reduction of wheat blast severity was obtained. The results revealed that the soil application of silicon, zinc and boron had a synergistic effect on the intensity of blast disease of wheat.

The present research work was conducted to evaluate the efficacy of eight fresh botanicals namely Azadirachta indica (Neem leaf), Allium cepa (Onion bulb), Allium sativum (Garlic), Curcuma longa (Turmeric), Zingiber officinale (Ginger), Allamanda cathartica (Allamanda leaf), Nigella sativa (Black cumin) and Aloe vera (Aloe vera) against Magnaporthe oryzae triticum (MoT) and solvent, ethanol (95%) was used for the phytochemical extraction of various plant parts. Three concentrations namely 1:1 (w/v), 1:0.50 (w/v) and 1:0.25 (w/v) of plant parts and ethanol were used for botanical extraction. The antifungal activity of botanicals against a virulent MoT isolate CHMoT07 was evaluated in-vitro using poison food technique. The lowest mycelia growth was recorded with Aloe vera (Aloe vera leaf) extracts and Nigella sativa (Black cumin seeds) extracts @ 1:1 w/v and @ 1:0.25 w/v with growth rate of 3.00 mm and 3.33 mm respectively at 7 days after inoculation, whereas the highest mycelia growth rate of MoT isolates was recorded in control plates both at 7 DAI and 14 DAI under in-vitro condition.

Wheat blast, caused by Magnaporthe oryzae, is considered a threat to wheat production globally. Yet, several epidemiological aspects of the disease are poorly understood. To elucidate these aspects, cross infection studies and molecular markers were utilized to identify alternate hosts (AH) that allow the permanence of M. oryzae pathotype Triticum in the wheat fields. Twenty-seven isolates from different weed species and six isolates from wheat were inoculated from the leaves of a blast susceptible wheat cultivar Caninde 11. Pot-2a, MoT3, Mo.T6098 and Mo.T6099 markers were used for the molecular diagnosis of the pathogen. Amplified fragments with the MoT3 marker were sequenced and aligned with the WB12 region used as reference sequence. The presence of M. oryzae pathotype Triticum on Brachiaria sp., Bromus catharticus, Lolium multiflorum, Avena strigosa and Eleusine indica is being reported for the first time. [ABSTRACT FROM AUTHOR]

Widespread resistance to QoIs, DMI and SDHIs fungicides has been reported for Brazilian populations of the wheat blast pathogen Pyricularia oryzae Triticum lineage (PoTl). A pre-existing resistance mechanism not associated with target site mutations has been indicated for resistance to DMIs and SDHIs, with strong indication that PoTl has multidrugresistance (MDR). Therefore, the main objective of this study was to test the hypothesis that resistance to DMI and SDHI fungicides detected in PoTl was due to efflux pump mediated MDR mechanism(s) by characterizing the sensitivity to antifungal efflux pump substrates. Four antifungal substrates were tested: tolnaftate (TOL), cycloheximide (CHX), rhodamine 6G (RH6G) and triphenyltin chloride (TPCL). TPCL and RH6G were considered the most relevant indicators for enhanced MDR activity. Among the 16 PoTl isolates tested, 9 were insensitive to TPCL, 1 to TOL, 16 to RH6G and 1 to CHX. The PoTl isolates were grouped into four distinct multidrug resistance phenotypes (MDRPs) based on resistance to combinations of fungicides and antifungal efflux pump substrates. Insensitivity to TPCL, RH6G and or TOL correlated well with DMI insensitivity, but MDR was not associated with SDHI resistance. The identification of multiple MDRP phenotypes associated with DMI resistance in our study warrants further research aimed at revealing the exact mechanisms of multidrug resistance in the wheat blast pathogen, including efflux pumps overexpression via transcriptomic analyses of differentially expressed genes; identification and discovery of mutations associated with changes in promoter regions or transcription factors of efflux transporters associated with multidrug resistance. [ABSTRACT FROM AUTHOR]

Epidemiological studies to better understand wheat blast (WB) spatial and temporal patterns were conducted in three field environments in Bolivia between 2019 and 2020. The temporal dynamics of wheat leaf blast (W L B) and spike blast (W S B) were best described by the logistic model compared with the Gompertz and exponential models. The nonlinear logistic infection rates were higher under defined inoculation in experiments two and three than under undefined inoculation in experiment one, and they were also higher for W S B than for W L B. The onset of W L B began with a spatial clustering pattern according to autocorrelation analysis and Moran's index values, with higher severity and earlier onset for defined than for undefined inoculation until the last sampling time. The W S B onset did not start with a spatial clustering pattern; instead, it was detected later until the last sampling date across experiments, with higher severity and earlier onset for defined than for undefined inoculation. Maximum severity ( K max ) was 1.0 for W S B and less than 1.0 for W L B. Aggregation of W L B and W S B was higher for defined than for undefined inoculation. The directionality of hotspot development was similar for both W L B and W S B, mainly occurring concentrically for defined inoculation. Our results show no evidence of synchronized development but suggest a temporal and spatial progression of disease symptoms on wheat leaves and spikes. Thus, we recommend that monitoring and management of WB should be considered during early growth stages of wheat planted in areas of high risk., Competing Interests: The author(s) declare no conflict of interest.

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 F 2 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 F 2:3 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., Competing Interests: The author(s) declare no conflict of interest.

Wheat blast, caused by the fungus Magnaporthe oryzae Triticum (MoT) pathotype, is a devastating disease persistent in South America and Bangladesh. Since MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced, disease control by fungicide application solely based on the detection of visual symptoms is ineffective. To develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control, we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments, MoT-6098 and MoT-6099, that are present in the MoT genome but not in the genome of the rice-infecting Magnaporthe oryzae Oryzae (MoO) pathotype. Using polymerase chain reaction (PCR), we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and Bangladesh. To test the efficiency of the two markers, we first established a loop-mediated isothermal amplification (LAMP) method to detect MoT at isothermal conditions, without the use of a PCR machine. Following this, we used the Cas12a protein and guide RNAs (gRNAs) to target the MoT-6098 and MoT-6099 sequences. The activated Cas12a showed indiscriminate single-stranded deoxyribonuclease (ssDNase) activity. We then combined target-dependent Cas12a ssDNase activation with recombinase polymerase amplification (RPA) and nucleic acid lateral flow immunoassay (NALFIA) to develop a method that accurately, sensitively, and cost-effectively detects MoT-specific DNA sequences in infected wheat plants. This novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.

The fundamental concepts of the genetics, race classification and epidemiology of the Wheat spike blast causing fungus Magnaporthe oryzae pathotype Triticum (MoT) are still evolving despite of its discovery in 1985 in Brazil for the first time. The fungus seems to defy the research progress that is being made globally by continuously evolving into pathotypes which have already overcome the much celebrated 2NS resistance in wheat lines as well as few of the initially effective fungicides. The compartmentalized i.e. two speed genome of the MoT, conferring the fungus an evolutionary advantage, has emerged as a challenge for the wheat spike blast researchers complicating its already difficult management. The airborne fungus with a range of alternative hosts is finding new geographical niches situated on different continents and is a matter of great apprehension among the nations whose food security is primarily dependent on wheat. The wheat blast outbreak in Bangladesh during 2016 was attributed to an isolate from Latin America escaping through a seed import consignment while the latest Zambian outbreak is still to be studied in detail regarding its origin and entry. The challenges in dealing wheat spike blast are not only on the level of genetics and epidemiology alone but also on the levels of policy making regarding international seed movement and research collaborations. The present review deals with these issues mainly concerning the effective management and controlling the international spread of this deadly disease of wheat, with a particular reference to India. We describe the origin, taxonomy, epidemiology and symptomology of MoT and briefly highlight its impact and management practices from different countries. We also discuss the advances in genomics and genome editing technologies that can be used to develop elite wheat genotypes resistant against different stains of wheat spike blast. [ABSTRACT FROM AUTHOR]

Protein kinases (PKs), being key regulatory enzymes of a wide range of signaling pathways, are potential targets for antifungal agents. Wheat blast disease, caused by Magnaporthe oryzae Triticum (MoT), is an existential threat to world food security. During the screening process of natural metabolites against MoT fungus, we find that two protein kinase inhibitors, staurosporine and chelerythrine chloride, remarkably inhibit MoT hyphal growth. This study further investigates the effects of staurosporine and chelerythrine chloride on MoT hyphal growth, conidia production, and development as well as wheat blast inhibition in comparison to a commercial fungicide, Nativo®75WG. The growth of MoT mycelia is significantly inhibited by these compounds in a dose-dependent manner. These natural compounds greatly reduce conidia production in MoT mycelia along with suppression of conidial germination and triggered lysis, resulting in deformed germ tubes and appressoria. These metabolites greatly suppress blast development in artificially inoculated wheat plants in the field. This is the first report of the antagonistic effect of these two natural PKC inhibitory alkaloids on MoT fungal developmental processes in vitro and suppression of wheat blast disease on both leaves and spikes in vivo. Further research is needed to identify their precise mechanism of action to consider them as biopesticides or lead compounds for controlling wheat blast. [ABSTRACT FROM AUTHOR]

Wheat blast, caused by the fungus Magnaporthe oryzae pathotype Triticum (MoT), constitutes one of the major obstacles to the expansion of wheat production in Bangladesh. In the absence of resistant variety, fungicide control is the first-hand effort. Determining an effective and economic fungicide spray schedule in controlling blast disease of wheat was aimed. Ten fungicides were tested during two consecutive cropping seasons of 2018-2019 to 2019-2020. The wheat plants of blast susceptible cultivar BARI Gom 26 were inoculated with spores (107 spores ml-1) of MoT at pre-heading stage of wheat (52 days age). Fungicides were applied both before inoculation and after the appearance of blast symptoms in cocktail for three times starting from booting of wheat at 7 days interval. Plants received the combination of Filia (Tricyclazole 40% + Propiconazole 12.5%) and Seltima (Pyraclostrobin 10%) had significantly lower blast incidence and severity (1.23% and 3.33%) against untreated plants. Cocktail of Nativo and Trooper (Tricyclazole 75 wp) proved 2nd best curative measure. Application of Nativo (Tebuconazole 50% + Trifloxystrobin 25%) alone ranked third in its efficacy. The fungicide spray schedule covered booting, pre-heading and heading stages of wheat. The results indicate a mixture of Tebuconazole + Tricyclazole + Pyraclostrobin is more effective (97% blast reduction) and economic (BCR 1.45) than a single compound application in reducing incidence and severity of wheat blast.

Wheat blast (WB) is a devastating fungal disease that has recently spread to Bangladesh and poses a threat to the wheat production in India, which is the second-largest wheat producing country in the world. In this study, 350 Indian wheat genotypes were evaluated for WB resistance in 12 field experiments in three different locations, namely Jashore in Bangladesh and Quirusillas and Okinawa in Bolivia. Single nucleotide polymorphisms (SNPs) across the genome were obtained using DArTseq® technology, and 7554 filtered SNP markers were selected for a genome-wide association study (GWAS). All the three GWAS approaches used identified the 2NS translocation as the only major source of resistance, explaining up to 32% of the phenotypic variation. Additional marker-trait associations were located on chromosomes 2B, 3B, 4D, 5A and 7A, and the combined effect of three SNPs (2B_180938790, 7A_752501634 and 5A_618682953) showed better resistance, indicating their additive effects on WB resistance. Among the 298 bread wheat genotypes, 89 (29.9%) carried the 2NS translocation, the majority of which (60 genotypes) were CIMMYT introductions, and 29 were from India. The 2NS carriers with a grand mean WB index of 6.6 showed higher blast resistance compared to the non-2NS genotypes with a mean index of 46.5. Of the 52 durum wheats, only one genotype, HI 8819, had the 2NS translocation and was the most resistant, with a grand mean WB index of 0.93. Our study suggests that the 2NS translocation is the only major resistance source in the Indian wheat panel analysed and emphasizes the urgent need to identify novel non-2NS resistance sources and genomic regions. [ABSTRACT FROM AUTHOR]

Abstract Wheat blast disease caused by a South American lineage of Magnaporthe oryzae Triticum (MoT) pathotype has emerged as a serious threat to wheat production in Bangladesh since its first emergence in 2016. Efficient and suitable methods for isolation, storage, inoculum production and molecular characterization of the pathogen can help in achieving the target of sustainable management of the disease in a relatively short period of time. In this study, we aimed to develop suitable methods for isolation, storage and morphological characterization and molecular identification of MoT isolates collected from the blast-infected wheat fields in Bangladesh. This process included modification of existing protocols that were available for a related fungal pathogen M. oryzae or de novo method development and validation. We developed suitable methods for isolation of MoT from field-infected plant samples using modified monoconidial isolation technique and produced abundant conidia from a single mycelial plate for in vivo pathogenicity assay in a reproducible manner. Cultural and morphological characterization of the isolates revealed that all Bangladeshi MoT isolates are of a single clonal lineage with similar cultural and morphological characters. Molecular detection of isolates with M. oryzae-specific primers Pot1 and Pot2 and MoT-specific primers MoT3F and MoT3R produced bands with the expected size from all wheat-infecting isolates. We also successfully established a PCR-based detection system based on a commercially available detection kit for field-infected leaf and seed samples by detecting Pot2- and MoT3-specific bands. Additionally, the simple method we developed in our study for producing abundant conidia in a very short period of time will be very helpful in studying biology of the wheat blast fungus. This method was also proven to be more user-friendly and cost-effective than previously available methods. Successful characterization of MoT isolates at morphological and molecular levels coupled with detection of the pathogen in infected field and seed lots should be useful for efficient surveillance and management of the fearsome wheat blast disease.

Abstract Background Wheat blast, caused by Magnaporthe oryzae Triticum (MoT) pathotype, is a global threat to wheat (Triticum aestivum L.) production. Few blast resistance (R) genes have been identified to date, therefore assessing potential sources of resistance in wheat is important. The Brazilian wheat cultivar BR 18-Terena is considered one of the best sources of resistance to blast and has been widely used in Brazilian breeding programmes, however the underlying genetics of this resistance are unknown. Results BR 18-Terena was used as the common parent in the development of two recombinant inbred line (RIL) F6 populations with the Brazilian cultivars Anahuac 75 and BRS 179. Populations were phenotyped for resistance at the seedling and heading stage using the sequenced MoT isolate BR32, with transgressive segregation being observed. Genetic maps containing 1779 and 1318 markers, were produced for the Anahuac 75 × BR 18-Terena and BR 18-Terena × BRS 179 populations, respectively. Five quantitative trait loci (QTL) associated with seedling resistance, on chromosomes 2B, 4B (2 QTL), 5A and 6A, were identified, as were four QTL associated with heading stage resistance (1A, 2B, 4A and 5A). Seedling and heading stage QTL did not co-locate, despite a significant positive correlation between these traits, indicating that resistance at these developmental stages is likely to be controlled by different genes. BR 18-Terena provided the resistant allele for six QTL, at both developmental stages, with the largest phenotypic effect conferred by a QTL being 24.8% suggesting that BR 18-Terena possesses quantitative resistance. Haplotype analysis of 100 Brazilian wheat cultivars indicates that 11.0% of cultivars already possess a BR 18-Terena-like haplotype for more than one of the identified heading stage QTL. Conclusions This study suggests that BR 18-Terena possesses quantitative resistance to wheat blast, with nine QTL associated with resistance at either the seedling or heading stage being detected. Wheat blast resistance is also largely tissue-specific. Identification of durable quantitative resistances which can be combined with race-specific R gene-mediated resistance is critical to effectively control wheat blast. Collectively, this work facilitates marker-assisted selection to develop new varieties for cultivation in regions at risk from this emerging disease.

Wheat is the second-largest cereal crop in Bangladesh and plays an essential role in ensuring the country's food security. Since 2016, there has been a severe epidemic of wheat blast disease in Bangladesh. This research investigated the nutritional context of wheat blast epidemics by analyzing the infected plants and allied soils. We collected blast-infected wheat plants and allied soil samples from six different severely infected regions of the Meherpur district situated in the western part of Bangladesh. The incidence and severity of wheat blast disease in the sampling fields ranged from 84.78 - 95.11% and 82.06–92.19%, respectively. Among the investigated mineral nutrients in plant samples, the concentrations of sulfur (S), calcium (Ca), magnesium (Mg), iron (Fe) and manganese (Mn) were within the acceptable range of the reference values. In contrast, 50% of the plant samples had insufficient phosphorus (P) concentrations, while others were within the critical range. The potassium (K) and copper (Cu) concentrations in more than 33.5% of plants were within the deficient range. The Si concentrations in half of the tested plant samples were below the acceptable level. However, the boron (B) concentration of around 50% of samples was within the toxic range. The total K, Ca, Zn, Fe, Mn, and Cu concentrations of the soils were lower than the reference values. Based on the interpretation of the available soil test values, the concentrations of S, Fe, Mn, and B in most samples were very low. The concentrations of available P, K, Ca, Mg, Zn and Cu in soil samples were higher than the critical limit. There was a negative relationship between K, S, Ca, Mg, Na and Si concentrations with blast incidence and severity. Therefore, this research suggests that certain plant nutrients such as P, K, Cu, B and Si play a vital role in the wheat blast disease epidemic.

Wheat blast caused by Pyricularia oryzae pathotype Triticum (MoT) has been transmitted from South America to Bangladesh and Zambia and is now spreading in these countries. To prepare against its further spread to Asian countries, we introduced Rmg8 , a gene for resistance to wheat blast, into a Japanese elite cultivar, Chikugoizumi (ChI), through recurrent backcrosses and established ChI near-isogenic lines, #2-1-10 with the Rmg8 / Rmg8 genotype and #4-2-10 with the rmg8 / rmg8 genotype. A molecular analysis suggested that at least 96.6% of the #2-1-10 genome was derived from the recurrent parent ChI. The #2-1-10 line was resistant to MoT not only in primary leaves at the seedling stage but also in spikes and flag leaves at the heading stage. The strength of the resistance in spikes of this Rmg8 carrier was comparable to that of a carrier of the 2NS segment, which has been the only genetic resource released to farmers' fields for wheat blast resistance. On the other hand, the 2NS resistance was not expressed on leaves at the seedling stage nor flag leaves at the heading stage. Considering that leaf blast has been increasingly reported and regarded as an important inoculum source for spike blast, Rmg8 expressed at both the seedling and heading stages, or more strictly in both leaves and spikes, is suggested to be useful to prevent the spread of MoT in Asia and Africa., Competing Interests: The author(s) declare no conflict of interest.

Despite advancements in agricultural research and the introduction of modern biotechnological and farming techniques, food security remains a significant issue. Although the efforts of farmers to meet the demands of a growing population, many plant diseases caused by pathogens, through their effects on cell division and tissue growth, lead to the annual loss of countless food crops. The recently emerged wheat blast fungus Magnaporthe oryzae pathotype Triticum (MoT ) poses a significant danger to worldwide wheat cultivation. The fungus is a highly varied lineage of the M. oryzae , responsible for causing rice blast disease. In spite of being a significant challenge to successful wheat production in South America since 1985, the underlying biology of the wheat blast pathogen is still not fully understood. The initial outbreak of the wheat blast in South Asia had a severe impact on wheat production, resulting in a complete loss of yield in affected fields. For the purpose of enhancing disease management, it's vital to acquire a comprehensive comprehension of the infection biology of the fungus and its interaction with wheat plants on molecular levels. Host-pathogen protein interactions (HPIs) have the potential to reveal the pathogens' mechanism for overcoming the host organism. The current study delves into the interactions between the host plant wheat and MoT through protein-protein interactions, molecular docking, and 100 ns molecular dynamic simulations. This research uncovers the structural and functional basis of these proteins, leading to improved plant health and production.Communicated by Ramaswamy H. Sarma.

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Wheat blast caused by Magnaporthe oryzae pathotype Triticum (MoT), initially restricted to South America, is a global threat for wheat after spreading to Asia in 2016 by the introduction of contaminated seeds, raising the question about transmission of the pathogen from seeds to seedlings, a process so far not well understood. We therefore studied the relationship between seed infection and disease symptoms on seedlings and adult plants. To accomplish this objective, we inoculated spikes of wheat cv. Apogee with a transgenic isolate (MoT‐DsRed, with the addition of being resistant to hygromycin). We identified MoT‐DsRed in experiments using hygromycin resistance for selection or by observation of DsRed fluorescence. The seeds from infected plants looked either apparently healthy or shrivelled. To evaluate the transmission, two experimental designs were chosen (blotter test and greenhouse) and MoT‐DsRed was recovered from both. This revealed that MoT is able to colonize wheat seedlings from infected seeds under the ground. The favourable conditions of temperature and humidity allowed a high recovery rate of MoT from wheat shoots when grown in artificial media. Around 42 days after germination of infected seeds, MoT‐DsRed could not be reisolated, indicating that fungal progression, at this time point, did not proceed systemically/endophytically. We hypothesize that spike infection might occur via spore dispersal from infected leaves rather than within the plant. Because MoT‐DsRed was not only successfully reisolated from seed coats and germinating seeds with symptoms, but also from apparently healthy seeds, urgent attention is needed to minimize the risks of inadvertent dispersal of inoculum. [ABSTRACT FROM AUTHOR]

Telomeres form the ends of linear chromosomes and usually comprise protein complexes that bind to simple repeated sequence motifs that are added to the 3′ ends of DNA by the telomerase reverse transcriptase (TERT). One of the primary functions attributed to telomeres is to solve the “end-replication problem” which, if left unaddressed, would cause gradual, inexorable attrition of sequences from the chromosome ends and, eventually, loss of viability. Telomere-binding proteins also protect the chromosome from 5′ to 3′ exonuclease action, and disguise the chromosome ends from the double-strand break repair machinery whose illegitimate action potentially generates catastrophic chromosome aberrations. Telomeres are of special interest in the blast fungus, Pyricularia, because the adjacent regions are enriched in genes controlling interactions with host plants, and the chromosome ends show enhanced polymorphism and genetic instability. Previously, we showed that telomere instability in some P. oryzae strains is caused by novel retrotransposons (MoTeRs) that insert in telomere repeats, generating interstitial telomere sequences that drive frequent, break-induced rearrangements. Here, we sought to gain further insight on telomeric involvement in shaping Pyricularia genome architecture by characterizing sequence polymorphisms at chromosome ends, and surrounding internalized MoTeR loci (relics) and interstitial telomere repeats. This provided evidence that telomere dynamics have played historical, and likely ongoing, roles in shaping the Pyricularia genome. We further demonstrate that even telomeres lacking MoTeR insertions are poorly preserved, such that the telomere-adjacent sequences exhibit frequent presence/absence polymorphism, as well as exchanges with the genome interior. Using TERT knockout experiments, we characterized chromosomal responses to failed telomere maintenance which suggested that much of the MoTeR relic-/interstitial telomere-associated polymorphism could be driven by compromised telomere function. Finally, we describe three possible examples of a phenomenon known as “Adaptive Telomere Failure,” where spontaneous losses of telomere maintenance drive rapid accumulation of sequence polymorphism with possible adaptive advantages. Together, our data suggest that telomere maintenance is frequently compromised in Pyricularia but the chromosome alterations resulting from telomere failure are not as catastrophic as prior research would predict, and may, in fact, be potent drivers of adaptive polymorphism.

Wheat blast (WB) caused by Magnaporthe oryzae pathotype Triticum (MoT) is an important fungal disease in tropical and subtropical wheat production regions. The disease was initially identified in Brazil in 1985, and it subsequently spread to some major wheat-producing areas of the country as well as several South American countries such as Bolivia, Paraguay, and Argentina. In recent years, WB has been introduced to Bangladesh and Zambia via international wheat trade, threatening wheat production in South Asia and Southern Africa with the possible further spreading in these two continents. Resistance source is mostly limited to 2NS carriers, which are being eroded by newly emerged MoT isolates, demonstrating an urgent need for identification and utilization of non-2NS resistance sources. Fungicides are also being heavily relied on to manage WB that resulted in increasing fungal resistance, which should be addressed by utilization of new fungicides or rotating different fungicides. Additionally, quarantine measures, cultural practices, non-fungicidal chemical treatment, disease forecasting, biocontrol etc., are also effective components of integrated WB management, which could be used in combination with varietal resistance and fungicides to obtain reasonable management of this disease.

Wheat blast is a threat to global wheat production, and limited blast-resistant cultivars are available. The current estimations of wheat spike blast severity rely on human assessments, but this technique could have limitations. Reliable visual disease estimations paired with Red Green Blue (RGB) images of wheat spike blast can be used to train deep convolutional neural networks (CNN) for disease severity (DS) classification. Inter-rater agreement analysis was used to measure the reliability of who collected and classified data obtained under controlled conditions. We then trained CNN models to classify wheat spike blast severity. Inter-rater agreement analysis showed high accuracy and low bias before model training. Results showed that the CNN models trained provide a promising approach to classify images in the three wheat blast severity categories. However, the models trained on non-matured and matured spikes images showing the highest precision, recall, and F1 score when classifying the images. The high classification accuracy could serve as a basis to facilitate wheat spike blast phenotyping in the future.

Telomeres form the ends of linear chromosomes and usually comprise protein complexes that bind to simple repeated sequence motifs that are added to the 3′ ends of DNA by the telomerase reverse transcriptase (TERT). One of the primary functions attributed to telomeres is to solve the "end-replication problem" which, if left unaddressed, would cause gradual, inexorable attrition of sequences from the chromosome ends and, eventually, loss of viability. Telomere-binding proteins also protect the chromosome from 5′ to 3′ exonuclease action, and disguise the chromosome ends from the double-strand break repair machinery whose illegitimate action potentially generates catastrophic chromosome aberrations. Telomeres are of special interest in the blast fungus, Pyricularia , because the adjacent regions are enriched in genes controlling interactions with host plants, and the chromosome ends show enhanced polymorphism and genetic instability. Previously, we showed that telomere instability in some P. oryzae strains is caused by novel retrotransposons (MoTeRs) that insert in telomere repeats, generating interstitial telomere sequences that drive frequent, break-induced rearrangements. Here, we sought to gain further insight on telomeric involvement in shaping Pyricularia genome architecture by characterizing sequence polymorphisms at chromosome ends, and surrounding internalized MoTeR loci (relics) and interstitial telomere repeats. This provided evidence that telomere dynamics have played historical, and likely ongoing, roles in shaping the Pyricularia genome. We further demonstrate that even telomeres lacking MoTeR insertions are poorly preserved, such that the telomere-adjacent sequences exhibit frequent presence/absence polymorphism, as well as exchanges with the genome interior. Using TERT knockout experiments, we characterized chromosomal responses to failed telomere maintenance which suggested that much of the MoTeR relic-/interstitial telomere-associated polymorphism could be driven by compromised telomere function. Finally, we describe three possible examples of a phenomenon known as "Adaptive Telomere Failure," where spontaneous losses of telomere maintenance drive rapid accumulation of sequence polymorphism with possible adaptive advantages. Together, our data suggest that telomere maintenance is frequently compromised in Pyricularia but the chromosome alterations resulting from telomere failure are not as catastrophic as prior research would predict, and may, in fact, be potent drivers of adaptive polymorphism. [ABSTRACT FROM AUTHOR]

The Triticum pathotype of Magnaporthe oryzae (syn. Pyricularia oryzae) causes wheat blast, which has recently spread to Asia. To assess the potential risk of wheat blast in rice-wheat growing regions, we investigated the pathogenicity of 14 isolates of P. oryzae on 32 wheat cultivars, among which Oryzae pathotype of P. oryzae (MoO) isolates were completely avirulent on the wheat cultivars at 22°C but caused various degrees of infection 25°C. These reactions at 25°C were isolate and cultivar dependent, like race-cultivar specificity, which was also recognized at the heading stage and caused typical blast symptoms on spikes. Microscopic analyses indicated that a compatible MoO isolate produced appressoria and infection hyphae on wheat as on rice. When we compared transcriptomes in wheat-MoO interactions, the bulk of pathogen-related genes were upregulated or downregulated in compatible and incompatible patterns, but changes in gene transcription were more significant in a compatible pattern. These results indicate that temperature could influence the infection ratio of wheat with MoO, and some MoO strains could be potential pathogens that increase the risk of wheat blast outbreaks in wheat-rice growing regions with global warming. In addition, certain wheat cultivars exhibited resistance and are assumed to carry resistance-promoting genes to the MoO strains. [ABSTRACT FROM AUTHOR]

Wheat blast (WB) caused by Magnaporthe oryzae pathotype Triticum (MoT) is an important fungal disease in tropical and subtropical wheat production regions. The disease was initially identified in Brazil in 1985, and it subsequently spread to some major wheat-producing areas of the country as well as several South American countries such as Bolivia, Paraguay, and Argentina. In recent years, WB has been introduced to Bangladesh and Zambia via international wheat trade, threatening wheat production in South Asia and Southern Africa with the possible further spreading in these two continents. Resistance source is mostly limited to 2NS carriers, which are being eroded by newly emerged MoT isolates, demonstrating an urgent need for identification and utilization of non-2NS resistance sources. Fungicides are also being heavily relied on to manage WB that resulted in increasing fungal resistance, which should be addressed by utilization of new fungicides or rotating different fungicides. Additionally, quarantine measures, cultural practices, non-fungicidal chemical treatment, disease forecasting, biocontrol etc., are also effective components of integrated WB management, which could be used in combination with varietal resistance and fungicides to obtain reasonable management of this disease. [ABSTRACT FROM AUTHOR]

A nursery comprising 25 elite bread wheat genotypes were evaluated to identify the sources of resistance to wheat blast caused by Magnaporthe oryzae pathotype Triticum (MoT), under artificial and natural infection experiments in three consecutive years from 2018 to 2020 in two different locations of Bangladesh. Seedling evaluation results revealed the absence of resistant and most genotypes displayed moderately susceptible to susceptible reaction. In the adult-plant field testing, six genotypes namely Borlaug 100, BARI Gom 33, BAW 1272, BAW 1280, BAW 1286, and BAW 1297 consistently showed resistant response to wheat head blast (WHB) infection with low levels of disease severity ranging from 0.2 to 8.1%. Among the remaining genotypes, some were moderately resistant, and the others exhibited various levels of susceptibility. The resistant genotypes that were most promising showed good agronomic performance under inoculated field trials. A significant positive correlation was observed between seedling and adult plant resistance. The above mentioned six genotypes also showed resistant reaction to WHB infection under controlled conditions when tested with three selected MoT isolates. The presence of 2NS/2AS translocation conferring WHB resistance was confirmed in four of the tested genotypes. Three (BAW 1272, BAW 1286, and BAW 1297) lines having non-2NS background exhibited good blast resistance under both the field and controlled conditions. These results will be useful for breeders and pathologists in exploiting the 2NS and non-2NS based resistant sources to pyramid and develop broad genetic based resistant germplasm. [ABSTRACT FROM AUTHOR]

Wheat blast is a threat to global wheat production, and limited blast-resistant cultivars are available. The current estimations of wheat spike blast severity rely on human assessments, but this technique could have limitations. Reliable visual disease estimations paired with Red Green Blue (RGB) images of wheat spike blast can be used to train deep convolutional neural networks (CNN) for disease severity (DS) classification. Inter-rater agreement analysis was used to measure the reliability of who collected and classified data obtained under controlled conditions. We then trained CNN models to classify wheat spike blast severity. Inter-rater agreement analysis showed high accuracy and low bias before model training. Results showed that the CNN models trained provide a promising approach to classify images in the three wheat blast severity categories. However, the models trained on non-matured and matured spikes images showing the highest precision, recall, and F1 score when classifying the images. The high classification accuracy could serve as a basis to facilitate wheat spike blast phenotyping in the future. [ABSTRACT FROM AUTHOR]

Wheat blast occurred in Bangladesh for the first time in Asia in 2016. It is caused by a fungal pathogen, Magnaporthe oryzae Triticum (MoT) pathotype. In this review, we focused on the current status of the wheat blast in regard to host, pathogen, and environment. Despite the many efforts to control the disease, it expanded to neighboring regions including India, the worlds second largest wheat producer. However, the disease occurrence has definitely decreased in quantity, because of many farmers chose to grow alternate crops according to the governments directions. Bangladesh government planned to introduce blast resistant cultivars but knowledges about genetics of resistance is limited. The genome analyses of the pathogen population revealed that the isolates caused wheat blast in Bangladesh are genetically close to a South American lineage of Magnaporthe oryzae. Understanding the genomes of virulent strains would be important to find target resistance genes for wheat breeding. Although the drier winter weather in Bangladesh was not favorable for development of wheat blast before, recent global warming and climate change are posing an increasing risk of disease development. Bangladesh outbreak in 2016 was likely to be facilitated by an extraordinary warm and humid weather in the affected districts before the harvest season. Coordinated international collaboration and steady financial supports are needed to mitigate the fearsome wheat blast in South Asia before it becomes a catastrophe.

Widespread resistance to QoIs, DMI and SDHIs fungicides has been reported for Brazilian populations of the wheat blast pathogen Pyricularia oryzae Triticum lineage (PoTl). A pre-existing resistance mechanism not associated with target site mutations has been indicated for resistance to DMIs and SDHIs, with strong indication that PoTl has multidrugresistance (MDR). Therefore, the main objective of this study was to test the hypothesis that resistance to DMI and SDHI fungicides detected in PoTl was due to efflux pump mediated MDR mechanism(s) by characterizing the sensitivity to antifungal efflux pump substrates. Four antifungal substrates were tested: tolnaftate (TOL), cycloheximide (CHX), rhodamine 6G (RH6G) and triphenyltin chloride (TPCL). TPCL and RH6G were considered the most relevant indicators for enhanced MDR activity. Among the 16 PoTl isolates tested, 9 were insensitive to TPCL, 1 to TOL, 16 to RH6G and 1 to CHX. The PoTl isolates were grouped into four distinct multidrug resistance phenotypes (MDRPs) based on resistance to combinations of fungicides and antifungal efflux pump substrates. Insensitivity to TPCL, RH6G and or TOL correlated well with DMI insensitivity, but MDR was not associated with SDHI resistance. The identification of multiple MDRP phenotypes associated with DMI resistance in our study warrants further research aimed at revealing the exact mechanisms of multidrug resistance in the wheat blast pathogen, including efflux pumps overexpression via transcriptomic analyses of differentially expressed genes; identification and discovery of mutations associated with changes in promoter regions or transcription factors of efflux transporters associated with multidrug resistance.

Protein kinases (PKs), being key regulatory enzymes of a wide range of signaling pathways, are potential targets for antifungal agents. Wheat blast disease, caused by Magnaporthe oryzae Triticum (MoT), is an existential threat to world food security. During the screening process of natural metabolites against MoT fungus, we find that two protein kinase inhibitors, staurosporine and chelerythrine chloride, remarkably inhibit MoT hyphal growth. This study further investigates the effects of staurosporine and chelerythrine chloride on MoT hyphal growth, conidia production, and development as well as wheat blast inhibition in comparison to a commercial fungicide, Nativo®75WG. The growth of MoT mycelia is significantly inhibited by these compounds in a dose-dependent manner. These natural compounds greatly reduce conidia production in MoT mycelia along with suppression of conidial germination and triggered lysis, resulting in deformed germ tubes and appressoria. These metabolites greatly suppress blast development in artificially inoculated wheat plants in the field. This is the first report of the antagonistic effect of these two natural PKC inhibitory alkaloids on MoT fungal developmental processes in vitro and suppression of wheat blast disease on both leaves and spikes in vivo. Further research is needed to identify their precise mechanism of action to consider them as biopesticides or lead compounds for controlling wheat blast.

Blast is caused by the host-specific lineages of the fungus Magnaporthe oryzae and is the most important destructive disease in major crop plants, including rice and wheat. The first wheat blast outbreak that occurred in Bangladesh in 2016 and the recent epidemic in Zambia were caused by the M. oryzae Triticum (MoT) pathotype, a fungal lineage belonging to M. oryzae. Although a few reported wheat cultivars show modest resistance to MoT, the patterns of genetic variation and diversity of this pathotype make it crucial to identify additional lines of resistant wheat germplasm. Nearly 40 rice blast resistant and susceptible genes have so far been cloned. Here, we used BLAST analysis to locate two rice blast susceptible genes in the wheat reference genome, bsr-d1 and bsr-k1, and identified six identical homologous genes located on subgenomes A, B, and D. We uncovered a total of 171 single nucleotide polymorphisms (SNPs) in an ethyl methanesulfonate (EMS)-induced population, with mutation densities ranging from 1/1107.1 to 1/230.7 kb through Targeting Induced Local Lesions IN Genomes (TILLING) by sequencing. These included 81 SNPs located in exonic and promoter regions, and 13 coding alleles that are predicted to have severe effects on protein function, including two pre-mature mutants that might affect wheat blast resistance. The loss-of-function alleles identified in this study provide insights into new wheat blast resistant lines, which represent a valuable breeding resource.