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.

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.

Pyricularia oryzae is a species complex that causes blast disease on more than 50 species of poaceous plants. Pyricularia oryzae has a worldwide distribution as a rice pathogen and in the last 30 years emerged as an important wheat pathogen in southern Brazil. We conducted phylogenetic analyses using 10 housekeeping loci for 128 isolates of P. oryzae sampled from sympatric populations of wheat, rice, and grasses growing in or near wheat fields. Phylogenetic analyses grouped the isolates into three major clades. Clade 1 comprised isolates associated only with rice and corresponds to the previously described rice blast pathogen P. oryzae pathotype Oryza (PoO). Clade 2 comprised isolates associated almost exclusively with wheat and corresponds to the previously described wheat blast pathogen P. oryzae pathotype Triticum (PoT). Clade 3 contained isolates obtained from wheat as well as other Poaceae hosts. We found that Clade 3 is distinct from P. oryzae and represents a new species, Pyricularia graminis-tritici (Pgt). No morphological differences were observed among these species, but a distinctive pathogenicity spectrum was observed. Pgt and PoT were pathogenic and highly aggressive on Triticum aestivum (wheat), Hordeum vulgare (barley), Urochloa brizantha (signal grass), and Avena sativa (oats). PoO was highly virulent on the original rice host (Oryza sativa), and also on wheat, barley, and oats, but not on signal grass. We conclude that blast disease on wheat and its associated Poaceae hosts in Brazil is caused by multiple Pyricularia species. Pyricularia graminis-tritici was recently found causing wheat blast in Bangladesh. This indicates that P. graminis-tritici represents a serious threat to wheat cultivation globally.

Fungicides have not been effective in controlling the wheat blast disease in Brazil. An earlier analysis of 179 isolates of Pyricularia oryzae Triticum lineage (PoTl) sampled from wheat fields across six populations in central‐southern Brazil during 2012 discovered a high level of resistance to strobilurin fungicides. Here we analysed azole resistance in the same strains based on EC50 measurements for tebuconazole and epoxiconazole. All six Brazilian populations of PoTl exhibited high resistance to both azoles, with in vitro EC50 values that were at least 35 to 50 times higher than the recommended field doses. We sequenced the CYP51A and CYP51B genes to determine if they were likely to play a role in the observed azole resistance. Although we found five distinct haplotypes in PoTl carrying four nonsynonymous substitutions in CYP51A, none of these substitutions were correlated with elevated EC50. CYP51B was sequenced for nine PoTl isolates, three each representing low, medium, and high tebuconazole EC50. Both PoTl CYP51A and CYP51B could complement yeast CYP51 function. All PoTl CYP51A‐expressing yeast transformants were less sensitive to triazoles than the PoTl CYP51B ones. Transformants expressing PoTl CYP51A haplotype H1 carrying the R158K substitution were not more resistant than those expressing PoTl CYP51A haplotype H5, which is synonymous to haplotype H6, found in triazole‐sensitive P. oryzae Oryza isolates from rice blast. Therefore, the reduced triazole sensitivity of wheat blast isolates compared to rice blast isolates appears to be associated with a non‐target‐site related resistance mechanism acquired after higher exposure to triazoles.

25The wheat blast disease has been a serious constraint for wheat production in Latin America 26 since the late 1980s. We used a population genomics analysis including 95 genome 27 sequences of the wheat blast pathogen Pyricularia graminis-tritici (Pygt) and other 28Pyricularia species to show that Pygt is a distinct, highly diverse pathogen species with a 29 broad host range. We assayed 11 neutral SSR loci in 526 Pygt isolates sampled from wheat 30 and other grasses distributed across the wheat-growing region of Brazil to estimate gene 31 flow, assess the importance of sexual reproduction, and compare the genetic structures of 32Pygt populations infecting wheat and nearby grasses. Our results suggest a mixed 33 reproductive system that includes sexual recombination as well as high levels of gene flow 34 among regions, including evidence for higher gene flow from grass-infecting populations and 35 into wheat-infecting populations than vice versa. The most common virulence groups were 36 shared between the grass-and wheat-infecting Pygt populations, providing additional 37 evidence for movement of Pygt between wheat fields and nearby grasses. Analyses of 38 fruiting body formation found that proto-perithecia and perithecia developed on senescing 39 stems of wheat and other grass hosts, suggesting that sexual reproduction occurs mainly 40 during the saprotrophic phase of the disease cycle on dead residues. Phalaris canariensis 41 (canarygrass) supported the fullest development of perithecia, suggesting it is a promising 42 candidate for identifying the teleomorph in the field. Based on these findings, we formulated 43 a more detailed disease cycle for wheat blast that includes an important role for grasses 44 growing near wheat fields. Our findings strongly suggest that widely grown pasture grasses 45 function as a major reservoir of wheat blast inoculum and provide a temporal and spatial 46 bridge that connects wheat fields across Brazil. 47. CC-BY-NC-ND 4.0 International license It is made available under a (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/203455 doi: bioRxiv preprint first posted online Oct. 16, 2017; 3 Author summary (200 words) 48 After the first wheat blast epidemic occurred in 1985 in Paraná, Brazil, the disease 49 spread to Bolivia, Argentina, and Paraguay, and was introduced into Bangladesh in 2016 50 followed by India in 2017. Wheat blast is caused by Pyricularia graminis-tritici (Pygt), a 51 highly diverse pathogen species related to the rice blast fungus P. oryzae, but with an 52 independent origin and a broader host range. We conducted a large scale contemporary 53 sampling of Pygt from symptomatic wheat and other grass species across Brazil and analyzed 54 the genetic structure of Pygt populations. Pygt populations on both wheat and other grasses 55 had high genotypic and virulence diversity, a genetic structure consistent with a ...

Wheat blast is one of the most important and devastating fungal diseases of wheat in South America, South‐east Asia, and now in southern Africa. The disease can reduce grain yield by up to 70% and is best controlled using integrated disease management strategies. The difficulty in disease management is compounded by the lack of durable host resistance and the ineffectiveness of fungicide sprays. New succinate dehydrogenase inhibitor (SDHI) fungicides were recently introduced for the management of wheat diseases. Brazilian field populations of the wheat blast pathogen Pyricularia oryzae Triticum lineage (PoTl) sampled from different geographical regions in 2012 and 2018 were shown to be resistant to both QoI (strobilurin) and DMI (azole) fungicides. The main objective of the current study was to determine the SDHI baseline sensitivity in these populations. Moderate levels of SDHI resistance were detected in five out of the six field populations sampled in 2012 and in most of the strains isolated in 2018. No association was found between target site mutations in the sdhB, sdhC, and sdhD genes and the levels of SDHI resistance, indicating that a pre‐existing resistance mechanism not associated with target site mutations is probably present in Brazilian wheat blast populations.

Yellow and black Sigatoka, caused by Mycosphaerella fijiensis and M. musicola, respectively, are the most important worldwide foliar diseases of bananas. Disease control is heavily dependent on intensive fungicide sprays, which increase selection pressure for fungicide resistance in pathogen populations. The primary objective of this study was to assess the level and spread of resistance to quinone-outside inhibitors (QoI—strobilurin) fungicides in populations of both pathogens sampled from banana fields under different fungicide spray regimes in Southeastern Brazil. Secondly, we aimed to investigate when QoI resistance was confirmed if this was associated with the target-site alteration G143A caused by a mutation in the mitochondrial encoded cytochrome b gene. QoI resistance was detected in fungicide treated banana fields, while no resistance was detected in the organic banana field. A total of 18.5% of the isolates sampled from the pathogens’ populations were resistant to QoI. The newly described M. thailandica was also found. It was the second most abundant Mycosphaerella species associated with Sigatoka-like leaf spot symptoms in the Ribeira Valley and the highest level of QoI resistance was found for this pathogen. The G143A cytochrome b alteration was associated with the resistance to the QoI fungicides azoxystrobin and trifloxystrobin in M. fijiensis, M. musicola and M. thailandica strains. In order to reduce resistance development and maintain the efficacy of QoI fungicides, anti-resistance management strategies based on integrated disease management practices should be implemented to control the Sigatoka disease complex.

ABSTRACT. We studied the effects of storage temperature on the activities of phenylalanine ammonialyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO) in minimally processed kale (Brassica oleracea var. acephala) that was stored for 15 and 9 days at 5 ± 1ºC and 10 ± 1ºC, respectively. The main visual evidence for quality loss in whole leaves was yellowing and loss of turgescence. Minimally processed leaves presented significant browning, indicating increased POD and PPO activities. The PAL activity in minimally processed leaves stored at 5ºC was fourfold higher than that of whole leaves after two days of storage. We showed that minimal processing influenced PAL, POD and PPO activities. The activity of all enzymes studied increased during storage, indicating that changes in phenolic metabolism play an important role in the decline of kale quality. PAL activity increased rapidly at the beginning of storage and exhibited a reduced rate of increase over time, while the PPO and POD activities increased continuously over time. The storage at 5 ° C was a great ally in delaying changes in phenolic metabolism; however, the absolute PAL activity was higher at 5 than at 10ºC.Keywords: Brassica oleracea cv. acephala, enzymatic browning, peroxidase, polyphenol oxidase, phenylalanine ammonia-lyase.Efeito da temperatura de conservação na qualidade e no metabolismo fenólico de folhas de couve inteiras e minimamente processadas RESUMO. Estudou-se o efeito da temperatura de armazenamento e do processamento mínimo sobre a atividade da fenilalanina amônia-liase (PAL), peroxidase (POD) e polifenol oxidase (PPO), em folhas de couve (Brassica oleracea var. acephala) por 15 e 9 dias, nas condições de 5 ± 1ºC e de 10 ± 1ºC, respectivamente. O amarelecimento e a perda de turgescência foram as principais causas para a redução da qualidade visual das folhas inteiras, enquanto nas folhas minimamente processadas apresentaram também, escurecimento, que coincidiu com o aumento na atividade da POD e PPO nas duas temperaturas estudadas. A atividade da PAL em folhas minimamente processadas mantidas a 5ºC mais que dobrou até o segundo dia de conservação em relação às conservadas a 10ºC. O processamento mínimo influenciou a atividade da PAL, POD e PPO. Todas as enzimas avaliadas, aumentaram a atividade durante a conservação, seja no início, no caso da PAL, ou no final, para a PPO e POD, indicando que as mudanças no metabolismo fenólico desempenha um papel importante no declínio da qualidade de couve. A conservação a 5ºC retardou as alterações no metabolismo fenólico, embora nessa temperatura, a atividade da PAL foi maior, em grandeza numérica, em relação às folhas armazenadas à 10ºC.Palavras-chave: Brassica oleraceae cv. acephala, escurecimento enzimático, peroxidase, polifenol oxidase, fenilalanina amônialiase.