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.

Emergence of wheat blast in Bangladesh was caused by a South American lineage of Magnaporthe oryzae.

BACKGROUND: In February 2016, a new fungal disease was spotted in wheat fields across eight districts in Bangladesh. The epidemic spread to an estimated 15,000 hectares, about 16 % of the cultivated wheat area in Bangladesh, with yield losses reaching up to 100 %. Within weeks of the onset of the epidemic, we performed transcriptome sequencing of symptomatic leaf samples collected directly from Bangladeshi fields. RESULTS: Reinoculation of seedlings with strains isolated from infected wheat grains showed wheat blast symptoms on leaves of wheat but not rice. Our phylogenomic and population genomic analyses revealed that the wheat blast outbreak in Bangladesh was most likely caused by a wheat-infecting South American lineage of the blast fungus Magnaporthe oryzae. CONCLUSION: Our findings suggest that genomic surveillance can be rapidly applied to monitor plant disease outbreaks and provide valuable information regarding the identity and origin of the infectious agent.

Development of genomic SSR markers and molecular characterization of Magnaporthe oryzae isolates from wheat in Brazil.

Magnaporthe oryzae, the causal agent of wheat blast, was characterized on a molecular level with 38 newly isolated genomic SSR loci. Among the 31 wheat isolates analyzed, 15 polymorphic loci were detected, with an average of 1.7 alleles per locus, 28.9% of them being highly or reasonably informative. The number of polymorphic loci was higher in isolates from Londrina in the Brazilian state of Paraná and Coromandel in Minas Gerais compared with Goiânia in Goiás and São Borja in Rio Grande do Sul. The rice isolate was clearly different from the wheat isolates, and the size difference in polymorphic SSR loci between one isolate from wheat and one isolate from rice was associated with the number of repeats. Some isolates collected from different states and in different years demonstrated similarities of 100%. The markers developed here are useful for the genetic analysis of M. oryzae isolated from wheat, and isolates representing the variability detected in the field can be used to search for better wheat blast resistance.

Identification of Fusarium head blight resistance loci in two Brazilian wheat mapping populations.

Fusarium head blight (FHB) is a disease of wheat (Triticum aestivum L.) that causes major yield losses in South America, as well as many other wheat growing regions around the world. FHB results in low quality, contaminated grain due to the production of mycotoxins such as deoxynivalenol (DON). In Brazil, FHB outbreaks are increasing in frequency and are currently controlled by fungicides which are costly and potentially harmful to the wider environment. To identify the genetic basis of resistance to FHB in Brazilian wheat, two mapping populations (Anahuac 75 × BR 18-Terena and BR 18-Terena × BRS 179) segregating for FHB resistance were phenotyped and quantitative trait loci (QTL) analysis was undertaken to identify genomic regions associated with FHB-related traits. A total of 14 QTL associated with FHB visual symptoms were identified, each of which explained 3.7-17.3% of the phenotypic variance. Two of these QTL were stable across environments. This suggests FHB resistance in Anahuac 75, BR 18-Terena and BRS 179 is controlled by multiple genetic loci that confer relatively minor differences in resistance. A major, novel QTL associated with DON accumulation was also identified on chromosome 4B (17.8% of the phenotypic variance), as well as a major QTL associated with thousand-grain weight on chromosome 6B (16.8% phenotypic variance). These QTL could be useful breeding targets, when pyramided with major sources of resistance such as Fhb1, to improve grain quality and reduce the reliance on fungicides in Brazil and other countries affected by FHB.

Wheat blast: from its origins in South America to its emergence as a global threat.

Wheat blast was first reported in Brazil in 1985. It spread rapidly across the wheat cropping areas of Brazil to become the most important biotic constraint on wheat production in the region. The alarming appearance of wheat blast in Bangladesh in 2016 greatly increased the urgency to understand this disease, including its causes and consequences. Here, we summarize the current state of knowledge of wheat blast and aim to identify the most important gaps in our understanding of the disease. We also propose a research agenda that aims to improve the management of wheat blast and limit its threat to global wheat production.

Wheat Blast: Past, Present, and Future.

The devastating wheat blast disease first emerged in Brazil in 1985. The disease was restricted to South America until 2016, when a series of grain imports from Brazil led to a wheat blast outbreak in Bangladesh. Wheat blast is caused by Pyricularia graminis-tritici ( Pygt), a species genetically distinct from the Pyricularia oryzae species that causes rice blast. Pygt has high genetic and phenotypic diversity and a broad host range that enables it to move back and forth between wheat and other grass hosts. Recombination is thought to occur mainly on the other grass hosts, giving rise to the highly diverse Pygt population observed in wheat fields. This review brings together past and current knowledge about the history, etiology, epidemiology, physiology, and genetics of wheat blast and discusses the future need for integrated management strategies. The most urgent current need is to strengthen quarantine and biosafety regulations to avoid additional spread of the pathogen to disease-free countries. International breeding efforts will be needed to develop wheat varieties with more durable resistance.

Gene Flow between Divergent Cereal- and Grass-Specific Lineages of the Rice Blast Fungus Magnaporthe oryzae.

Delineating species and epidemic lineages in fungal plant pathogens is critical to our understanding of disease emergence and the structure of fungal biodiversity and also informs international regulatory decisions. Pyricularia oryzae (syn. Magnaporthe oryzae) is a multihost pathogen that infects multiple grasses and cereals, is responsible for the most damaging rice disease (rice blast), and is of growing concern due to the recent introduction of wheat blast to Bangladesh from South America. However, the genetic structure and evolutionary history of M. oryzae, including the possible existence of cryptic phylogenetic species, remain poorly defined. Here, we use whole-genome sequence information for 76 M. oryzae isolates sampled from 12 grass and cereal genera to infer the population structure of M. oryzae and to reassess the species status of wheat-infecting populations of the fungus. Species recognition based on genealogical concordance, using published data or extracting previously used loci from genome assemblies, failed to confirm a prior assignment of wheat blast isolates to a new species (Pyricularia graminis-tritici). Inference of population subdivisions revealed multiple divergent lineages within M. oryzae, each preferentially associated with one host genus, suggesting incipient speciation following host shift or host range expansion. Analyses of gene flow, taking into account the possibility of incomplete lineage sorting, revealed that genetic exchanges have contributed to the makeup of multiple lineages within M. oryzae These findings provide greater understanding of the ecoevolutionary factors that underlie the diversification of M. oryzae and highlight the practicality of genomic data for epidemiological surveillance in this important multihost pathogen.IMPORTANCE Infection of novel hosts is a major route for disease emergence by pathogenic microorganisms. Understanding the evolutionary history of multihost pathogens is therefore important to better predict the likely spread and emergence of new diseases. Magnaporthe oryzae is a multihost fungus that causes serious cereal diseases, including the devastating rice blast disease and wheat blast, a cause of growing concern due to its recent spread from South America to Asia. Using whole-genome analysis of 76 fungal strains from different hosts, we have documented the divergence of M. oryzae into numerous lineages, each infecting a limited number of host species. Our analyses provide evidence that interlineage gene flow has contributed to the genetic makeup of multiple M. oryzae lineages within the same species. Plant health surveillance is therefore warranted to safeguard against disease emergence in regions where multiple lineages of the fungus are in contact with one another.

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.

Relationship between blast severity on seedlings and panicles of Brazilian oat cultivars / Relação entre a severidade de brusone em plântulas e panículas de cultivares brasileiras de aveia

ABSTRACT: Blast, caused by Pyricularia oryzae, threats black (Avena strigosa) and white (A. sativa) oats in Brazil. Little is known about the reaction of Brazilian oat cultivars to blast and if there is a relationship between the response of cultivars to the disease at seedling and adult plant stages. The goals of this research were to verify (a) the reaction of black and white Brazilian oat cultivars to infection of P. oryzae and (b) if the response to blast on seedling leaves and panicles was correlated. Seedlings and adult plants of 13 oat cultivars were inoculated with a conidial suspension of one P. oryzae isolate obtained from a black oat cultivar. The cultivars were classified according to blast severity on the leaf seedlings and panicles estimated by visual estimation. Two experiments were conducted for each one of the two evaluated stages. Cultivars with lower value to blast severity on seedling leaves were 'UPFA Gaudéria' (25.82%) and 'UPF 18' (24.88%) and, on panicles, 'BRS Centauro' (31.63%). Coefficients of Pearson, Spearman, and Kendall obtained from correlation analyses between blast severity on seedling leaves and panicles were -0.21232, -0.35714, and -0.30769, respectively (non-significant at 0.05). Lack of correlation between the responses of these oat cultivars to blast on seedling leaves and panicles be considered in the management of the disease in the field and the generation of new oat cultivars.

RESUMO: A brusone, causada por Pyricularia oryzae, ameaça as culturas da aveia preta (Avena strigosa) e branca (A. sativa) no Brasil. Sabe-se pouco sobre a reação das cultivares brasileiras de aveia à brusone e se existe relação entre a resposta destas cultivares à doença nos estádios de plântula e de planta adulta. Os objetivos deste trabalho foram: (a) verificar a reação de cultivares brasileiras de aveia à infecção de P. oryzae e; (b) se existe relação entre a resposta à brusone dessas cultivares nos estádios de plântula e de planta adulta. Plântulas e plantas adultas de 13 cultivares de aveia foram inoculadas com uma suspensão de conídios de um isolado de P. oryzae obtido de uma planta de uma cultivar de aveia preta. As cultivares foram classificadas de acordo com a estimativa visual da severidade da doença nas folhas das plântulas e nas panículas das plantas adultas. Dois experimentos foram conduzidos para cada um dos estádios avaliados. As cultivares com menor severidade de brusone nas plântulas foram a 'Gaudéria UPFA' (25,82%) e a 'UPF 18' (24,88%) e, nas panículas, foi a 'BRS Centauro' (31,63%). Os coeficientes de Pearson, Spearman e Kendall obtidos da análise de correlação entre os dois estádios avaliados foram -0,21232, -0,35714 e -0,30769, respectivamente (todos sem significância estatística à 0,05). A falta de correlação entre as respostas das cultivares de aveia à brusone, em plântulas e panículas, constitui-se em informação a ser considerada no manejo da doença, no campo e na geração de novas cultivares de aveia.

Resultados agronômicos e qualitativos da nova cultivar de trigo 'BRS Marcante' / Agronomic and qualitative results of the new wheat cultivar 'BRS Marcante'

A cultivar de trigo 'BRS Marcante' foi desenvolvida pela Embrapa, envolvendo um híbrido F1 do cruzamento entre as linhagens PF 980533 e PF 970227 com a cultivar 'BRS Guamirim', realizado em telado, na Embrapa Trigo, no inverno de 2003. As gerações segregantes foram conduzidas pelo método genealógico e a linhagem resultante, nomeada de PF 080310. A cultivar caracteriza-se pela sua ampla capacidade de adaptação às condições de cultivo do sul do Brasil, pelo bom potencial de rendimento de grãos e qualidade industrial da classe Pão.

The wheat cultivar 'BRS Marcante' was developed by Embrapa, as a result a cross between F1 hybrid between lines PF 980533 and PF 970227 with 'BRS Guamirim' and carried out in a green-house of Embrapa Wheat, on 2003 winter season. The segregate generations were conducted by genealogic method and the genotype resulted was named PF 080310. It has wide adaptation ability to south Brazilian conditions, high grain yield potential and belongs to Bread Class.

Esporulação de Pyricularia grisea em diferentes meios de cultura e regimes de luz / Sporulation of Pyricularia grisea on different culture media and light regimes

Diferentes fatores influenciam o crescimento e a esporulação dos fungos, tais como: composição do meio de cultivo, temperatura e luz. O objetivo do trabalho foi determinar o melhor meio de cultivo e regime de luz para a esporulação de Pyricularia grisea. Um isolado deste, obtido da cultivar de trigo 'BRS 194', foi repicado por disco micelial em oito meios de cultivo (farinha integral de arroz, aveia, centeio, cevada, trigo, triticale; BSA e extrato de tomate) e submetido a três regimes de luz (escuro, fotoperíodo de 12h e luz contínua). As placas permaneceram por 10 dias, em câmara de crescimento sob temperatura de 23-25°C. Em seguida, o número de conídios foi determinado em câmara de Neubauer. O meio de cultivo à base de farinha integral de trigo propiciou a maior esporulação do fungo em todos os regimes de luz, e a maior esporulação ocorreu em regime de luz contínua para todos os meios de cultivos testados. Os meios de arroz e BSA propiciaram uma menor produção de conídios pelo fungo.

A set of conditions is necessary for the growth and sporulation of fungi, particularly the composition of culture media, temperature and light. The objective of this research was to determine the ideal culture medium and light regime for the sporulation of P. grisea. Micelial discs of an isolate, obtained from cultivar 'BRS 194' of wheat, were transferred to eight culture media (rice wholemeal, oat, rye, barley, wheat, triticale, PSA and tomato sauce) and kept in the dark, under continuous light and 12h of light. In a growing chamber, for 10 days at 23-25°C. Then, the number of conidia was determined in a Neubauer chamber. The higher sporulation, regardless of light regime, was observed on wheat wholemeal culture medium. The continuous light provided higher sporulation this fungus in all culture medium. The lowest production of conidia was observed on rice wholemeal and PSA culture media.

Densidade populacional de Ralstonia solanacearum em cultivares de batata a campo / Population densities of Ralstonia solanacearum on potato cultivars in the field

A ocorrência de populaçöes latentes de Ralstonia solanacearum em plantas de batata (Solanum tuberosum) pode representar fonte de inóculo de potencial desconhecido. Além disso, também é desconhecido se a populaçäo latente da bactéria é menor em cultivares resistentes do que em cultivares suscetíveis. Com a finalidade de estudar estes aspectos, um experimento foi conduzido a campo em dois locais no Estado do Rio Grande do Sul, Brasil. O objetivo foi verificar se havia relaçäo entre a densidade populacional e o grau de resistência de cultivares de batata. O experimento foi conduzido em Eldorado do Sul, durante o período de primavera, e em Caxias do Sul, durante o período de outono. Tubérculos das cultivares Achat, Baronesa, Elvira, Macaca, Monte Bonito e Trapeira foram inoculados com uma estirpe de R. solanacearum, biovar II, e plantados a campo. A densidade populacional da bactéria foi estimada em plantas com e sem sintomas de murcha, através de ELISA e imunofluorescência. Näo houve evidência da relaçäo entre densidade populacional da bactéria e cultivar. Além disso, a densidade populacional na cultivar Achat, caracterizada como a mais resistente entre as cultivares testadas, foi igual à registrada nas outras cultivares.