Effect of Initial Inoculum on the Temporal and Spatial Dynamics of Wheat Blast Under Field Conditions in Bolivia.

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 (WLB) and spike blast (WSB) 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 WSB than for WLB. The onset of WLB 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 WSB 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 (Kmax) was 1.0 for WSB and less than 1.0 for WLB. Aggregation of WLB and WSB was higher for defined than for undefined inoculation. The directionality of hotspot development was similar for both WLB and WSB, 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.

Pyricularia Are Mostly Host-Specialized with Limited Reciprocal Cross-Infection Between Wheat and Endemic Grasses in Minas Gerais, Brazil.

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 Urochloa-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 cross-infection would be sufficient to provide an inoculum reservoir, or serve as a bridge between wheat growing regions, is questionable and, therefore, deserves further investigation.

Occurrence of Pyricularia oryzae Triticum in plants of the genus Urochloa in Brazil / Ocorrência de Pyricularia oryzae Triticum em plantas do gênero Urochloa no Brasil

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.

Neste estudo Pyricularia spp., P. oryzae e o patótipo Triticum (PoT) de P. oryzae foram detectados e identificados em segmentos foliares de forrageiras e gramíneas invasoras de lavouras de trigo. Em 2018 e 2019, foram coletadas 66 amostras de segmentos foliares lesionados de Urochloa e outras gramíneas em Londrina (PR), Patos de Minas (MG) e Uberaba (MG). A detecção e/ou identificação dos patógenos nas amostras foi realizada por meio de procedimentos de câmara úmida e com os iniciadores MoT3 e Pot2 por PCR. Houve amplificações de DNA com o primer MoT3 (específico para PoT) em 13 (19,69%) das amostras, todas provenientes da Urochloa. O resultado de que Urochloa hospeda PoT em uma taxa relativamente alta levanta preocupações sobre a importância que essas plantas podem ter no ciclo de brusone do trigo como hospedeiro intermediário para o patógeno e / ou fonte de inóculo para a doença.

Occurrence of Pyricularia oryzae Triticum in plants of the genus Urochloa in Brazil / Ocorrência de Pyricularia oryzae Triticum em plantas do gênero Urochloa no Brasil

ABSTRACT: 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.

RESUMO: Neste estudo Pyricularia spp., P. oryzae e o patótipo Triticum (PoT) de P. oryzae foram detectados e identificados em segmentos foliares de forrageiras e gramíneas invasoras de lavouras de trigo. Em 2018 e 2019, foram coletadas 66 amostras de segmentos foliares lesionados de Urochloa e outras gramíneas em Londrina (PR), Patos de Minas (MG) e Uberaba (MG). A detecção e/ou identificação dos patógenos nas amostras foi realizada por meio de procedimentos de câmara úmida e com os iniciadores MoT3 e Pot2 por PCR. Houve amplificações de DNA com o primer MoT3 (específico para PoT) em 13 (19,69%) das amostras, todas provenientes da Urochloa. O resultado de que Urochloa hospeda PoT em uma taxa relativamente alta levanta preocupações sobre a importância que essas plantas podem ter no ciclo de brusone do trigo como hospedeiro intermediário para o patógeno e / ou fonte de inóculo para a doença.

Higher fitness and competitive advantage of Pyricularia oryzae Triticum lineage resistant to QoI fungicides.

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.

Wheat Blast: A Disease Spreading by Intercontinental Jumps and Its Management Strategies.

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.

Suppression of wheat blast resistance by an effector of Pyricularia oryzae is counteracted by a host specificity resistance gene in wheat.

Wheat blast caused by the Triticum pathotype of Pyricularia oryzae poses a serious threat to wheat production in South America and Asia and is now becoming a pandemic disease. Here, we show that Rmg8, a promising wheat gene for resistance breeding, is suppressed by PWT4, an effector gene of P. oryzae, and in turn that the suppression is counteracted by Rwt4, a wheat gene recognizing PWT4. When PWT4 was introduced into a wheat blast isolate carrying AVR-Rmg8 (an avirulence gene corresponding to Rmg8), PWT4 suppressed wheat resistance conferred by Rmg8. PWT4 did not alter the expression of AVR-Rmg8, but higher expression of PWT4 led to more efficient suppression. This suppression was observed in rwt4 carriers, but not in Rwt4 carriers, indicating that it is counteracted by Rwt4. PWT4 was assumed to have been horizontally transferred from a weed-associated cryptic species, P. pennisetigena, to an Avena isolate of P. oryzae in Brazil. This implies a potential risk of the acquisition of PWT4 by the wheat blast fungus and the 'breakdown' of Rmg8. We suggest that Rmg8 should be introduced together with Rwt4 into a wheat cultivar when it is used for resistance breeding.

Effectiveness of the Wheat Blast Resistance Gene Rmg8 in Bangladesh Suggested by Distribution of an AVR-Rmg8 Allele in the Pyricularia oryzae Population.

Wheat blast caused by the Triticum pathotype of Pyricularia oryzae was first reported in 1985 in Brazil and recently spread to Bangladesh. We tested whether Rmg8 and RmgGR119, recently identified resistance genes, were effective against Bangladeshi isolates of the pathogen. Common wheat accessions carrying Rmg8 alone (IL191) or both Rmg8 and RmgGR119 (GR119) were inoculated with Brazilian isolates (Br48, Br5, and Br116.5) and Bangladeshi isolates (T-108 and T-109). Br48, T-108, and T-109 carried the eI type of AVR-Rmg8 (the avirulence gene corresponding to Rmg8) while Br5 and Br116.5 carried its variants, eII and eII' types, respectively. Detached primary leaves of IL191 and GR119 were resistant to all isolates at 25°C. At a higher temperature (28°C), their resistance was still effective against the eI carriers but was reduced to a low level against the eII/eII' carriers. A survey of databases and sequence analyses revealed that all Bangladeshi isolates carried the eI type which induced a higher level of resistance than the eII/eII' types. The resistance of IL191 (Rmg8/-) to the eI carriers was maintained even at the heading stage and at the higher temperature. In addition, GR119 (Rmg8/RmgGR119) displayed higher levels of resistance than IL191 at this stage. These results suggest that Rmg8 combined with RmgGR119 will be useful in breeding for resistance against wheat blast in Bangladesh.

Dissecting the genetic basis of wheat blast resistance in the Brazilian wheat cultivar BR 18-Terena.

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.

Conidia sporulation of Pyricularia oryzae in segments of wheat plants under six different temperatures / Esporulação de conídios de Pyricularia oryzae em segmentos de plantas de trigo sob seis diferentes temperaturas

Wheat blast is known for developing itself more intensely under relatively high temperature conditions but many aspects related to its epidemiology remain unknown. The objective of this research was to evaluate the sporulative capacity of Pyricularia oryzae Triticum (Pot), the causal agent of wheat blast, in tissues of wheat plants under different temperatures degrees. Wheat plants of the cultivar Anahuac 75, susceptible to blast, were inoculated in the stage of flowering with conidial suspensions (105 conidia/mL) of the Pot isolates Py 12.1.209 and Py 12.1.132. Seven days after the inoculation, plants were cut in the following segments: leaves, stems and rachis (with blast severity ranging from 40 to 60%). Groups of each one of the three plant segments with the lesions were disposed in Petri-dish moist chambers, that were submitted to six different temperature treatments (18, 21, 24, 27, 30 and 33 °C). The most appropriate model that related the conidia production with temperature was identified in the evaluations conducted with stems. The established equations allowed identifying that the highest production of conidia of Pot occurs between 24 and 27 °C.(AU)

Sabe-se que a brusone do trigo ocorre preferencialmente em condições de temperaturas relativamente altas, porém muitos aspectos sobre a epidemiologia desta doença ainda permanecem desconhecidos. O objetivo deste trabalho foi avaliar a capacidade esporulativa de Pyricularia oryzae Triticum (Pot) em tecidos de plantas de trigo sob diferentes temperaturas. Plantas de trigo do cultivar Anahuac 75, suscetível à brusone, foram inoculadas no estádio do florescimento com suspensões de conídios dos isolados Py 12.1.209 e Py 12.1.132 de Pot (105 conídios/mL). Sete dias após a inoculação, as plantas foram cortadas e separadas em três tipos de segmentos: colmos, folhas e ráquis (com severidade de brusone variando de 40 a 60%). Um grupo de cada desses três tipos de segmentos de planta contendo lesões foi disposto em câmara úmida. O material foi acondicionado em seis níveis de temperatura: 18, 21, 24, 27, 30 e 33 °C. O modelo mais apropriado que relacionou a produção de conídios com temperatura foi identificado nas avaliações feitas nas amostras de colmo. As equações estabelecidas permitiram identificar que a maior produção de conídios de Pot ocorreu entre as temperaturas de 24 e 27 °C.(AU)

Temporal Dynamics of Wheat Blast Epidemics and Disease Measurements Using Multispectral Imagery.

Wheat blast is a devastating disease caused by the Triticum pathotype of Magnaporthe oryzae. M. oryzae Triticum is capable of infecting leaves and spikes of wheat. Although symptoms of wheat spike blast (WSB) are quite distinct in the field, symptoms on leaves (WLB) are rarely reported because they are usually inconspicuos. Two field experiments were conducted in Bolivia to characterize the change in WLB and WSB intensity over time and determine whether multispectral imagery can be used to accurately assess WSB. Disease progress curves (DPCs) were plotted from WLB and WSB data, and regression models were fitted to describe the nature of WSB epidemics. WLB incidence and severity changed over time; however, the mean WLB severity was inconspicuous before wheat began spike emergence. Overall, both Gompertz and logistic models helped to describe WSB intensity DPCs fitting classic sigmoidal shape curves. Lin's concordance correlation coefficients were estimated to measure agreement between visual estimates and digital measurements of WSB intensity and to estimate accuracy and precision. Our findings suggest that the change of wheat blast intensity in a susceptible host population over time does not follow a pattern of a monocyclic epidemic. We have also demonstrated that WSB severity can be quantified using a digital approach based on nongreen pixels. Quantification was precise (0.96 < r> 0.83) and accurate (0.92 < ρ > 0.69) at moderately low to high visual WSB severity levels. Additional sensor-based methods must be explored to determine their potential for detection of WLB and WSB at earlier stages.

A Genomic Approach to Develop a New qPCR Test Enabling Detection of the Pyricularia oryzae Lineage Causing Wheat Blast.

Rapid detection is key to managing emerging diseases because it allows their spread around the world to be monitored and limited. The first major wheat blast epidemics were reported in 1985 in the Brazilian state of Paraná. Following this outbreak, the disease quickly spread to neighboring regions and countries and, in 2016, the first report of wheat blast disease outside South America was released. This Asian outbreak was due to the trade of infected South American seed, demonstrating the importance of detection tests in order to avoid importing contaminated biological material into regions free from the pathogen. Genomic analysis has revealed that one particular lineage within the fungal species Pyricularia oryzae is associated with this disease: the Triticum lineage. A comparison of 81 Pyricularia genomes highlighted polymorphisms specific to the Triticum lineage, and this study developed a real-time PCR test targeting one of these polymorphisms. The test's performance was then evaluated in order to measure its analytical specificity, analytical sensitivity, and robustness. The C17 quantitative PCR test detected isolates belonging to the Triticum lineage with high sensitivity, down to 13 plasmid copies or 1 pg of genomic DNA per reaction tube. The blast-based approach developed here to study P. oryzae can be transposed to other emerging diseases.

Conidia sporulation of Pyricularia oryzae in segments of wheat plants under six different temperatures / Esporulação de conídios de Pyricularia oryzae em segmentos de plantas de trigo sob seis diferentes temperaturas

ABSTRACT: Wheat blast is known for developing itself more intensely under relatively high temperature conditions but many aspects related to its epidemiology remain unknown. The objective of this research was to evaluate the sporulative capacity of Pyricularia oryzae Triticum (Pot), the causal agent of wheat blast, in tissues of wheat plants under different temperatures degrees. Wheat plants of the cultivar Anahuac 75, susceptible to blast, were inoculated in the stage of flowering with conidial suspensions (105 conidia/mL) of the Pot isolates Py 12.1.209 and Py 12.1.132. Seven days after the inoculation, plants were cut in the following segments: leaves, stems and rachis (with blast severity ranging from 40 to 60%). Groups of each one of the three plant segments with the lesions were disposed in Petri-dish moist chambers, that were submitted to six different temperature treatments (18, 21, 24, 27, 30 and 33 °C). The most appropriate model that related the conidia production with temperature was identified in the evaluations conducted with stems. The established equations allowed identifying that the highest production of conidia of Pot occurs between 24 and 27 °C.

RESUMO: Sabe-se que a brusone do trigo ocorre preferencialmente em condições de temperaturas relativamente altas, porém muitos aspectos sobre a epidemiologia desta doença ainda permanecem desconhecidos. O objetivo deste trabalho foi avaliar a capacidade esporulativa de Pyricularia oryzae Triticum (Pot) em tecidos de plantas de trigo sob diferentes temperaturas. Plantas de trigo do cultivar Anahuac 75, suscetível à brusone, foram inoculadas no estádio do florescimento com suspensões de conídios dos isolados Py 12.1.209 e Py 12.1.132 de Pot (105 conídios/mL). Sete dias após a inoculação, as plantas foram cortadas e separadas em três tipos de segmentos: colmos, folhas e ráquis (com severidade de brusone variando de 40 a 60%). Um grupo de cada desses três tipos de segmentos de planta contendo lesões foi disposto em câmara úmida. O material foi acondicionado em seis níveis de temperatura: 18, 21, 24, 27, 30 e 33 °C. O modelo mais apropriado que relacionou a produção de conídios com temperatura foi identificado nas avaliações feitas nas amostras de colmo. As equações estabelecidas permitiram identificar que a maior produção de conídios de Pot ocorreu entre as temperaturas de 24 e 27 °C.