Snow mold is a severe plant disease caused by psychrophilic or psychrotolerant fungi, of which Microdochium species are the most harmful. A clear understanding of Microdochium biology has many gaps; the pathocomplex and its dynamic are poorly characterized, virulence factors are unknown, genome sequences are not available, and the criteria of plant snow mold resistance are not elucidated. Our study aimed to identify comprehensive characteristics of a local community of snow mold-causing Microdochium species colonizing a particular crop culture. By using the next-generation sequencing (NGS) technique, we characterized fungal and bacterial communities of pink snow mold-affected winter rye (Secale cereale) plants within a given geographical location shortly after snowmelt. Twenty-one strains of M. nivale were isolated, classified on the basis of internal transcribed spacer 2 (ITS2) region, and characterized by morphology, synthesis of extracellular enzymes, and virulence. Several types of extracellular enzymatic activities, the level of which had no correlations with the degree of virulence, were revealed for Microdochium species for the first time. Our study shows that genetically and phenotypically diverse M. nivale strains simultaneously colonize winter rye plants within a common area, and each strain is likely to utilize its own, unique strategy to cause the disease using “a personal” pattern of extracellular enzymes.
Winter rye is a versatile crop widely used for food and industry. Although rye is resistant to abiotic stressors and many phytopathogens, it is severely damaged by pink snow mold (SM)—a progressive disease caused by the psychrotolerant fungus Microdochium nivale under the snow cover or during prolonged periods of wet and cool conditions. Due to little use of the SM resistance sources in contemporary breeding, varieties with at least moderate resistance to SM are limited. Our study aimed to integrate field assessment under natural conditions and an artificially enriched infection background with laboratory techniques for testing rye accessions and selecting SM resistant sources for applied breeding programs and genetic research. We revealed valuable sources of SM resistance and split rye accessions, according to the level of the genetic divergence of the SM resistance phenotype. This allowed us to select the most distinct donors of the SM resistance, for their use as parental forms, to include novel variability sources in the breeding program for achieving high genetic variability, as well as enhanced and durable SM resistance, in progeny. The rye accessions analyzed here, and the suggested options for their use in breeding, are valuable tools for rye breeding.
Our study investigates the intra- and interpopulation diversity of phytopathogenic fungi, using Microdochium nivale as a model organism. A total of 136 strains, representing two populations, were collected from different winter cereal crops (rye, wheat, and triticale) across two agrocenoses. In these strains, we analyzed and compared genetic and phenotypic traits, exploring potential relationships between them. Significant genetic and phenotypic diversity was revealed among the assayed strains, with most exhibiting distinctive genetic features compared to previously characterized strains from other countries. Both populations included highly virulent and lowly virulent strains, with the majority displaying similar virulence levels across different cereal crops. Some strains demonstrated differential virulence toward different crops, and some even exhibited a plant-beneficial effect on rye while being virulent to wheat and triticale. The percentages of strains with specific genetic traits were different in the two analyzed populations. Our study lays the groundwork for identifying genetic markers associated with phenotypic traits in the studied strains, which will aid in developing reliable diagnostics for predicting crop loss risk and improving disease management. Furthermore, it will assist in identifying reliable sources of resistance to M. nivale-induced diseases within the winter cereal gene pool.
The study investigated the adaptation of the snow mold causal fungus, Microdochium nivale, to the fungicides fludioxonil and tebuconazole. Analysis of intrapopulation diversity among 136 M. nivale strains from two Russian populations revealed no strains with high-level resistance to these fungicides. However, the strains exhibited considerable variability in their sensitivity to small fungicide doses. Fungicide sensitivity levels were not associated with virulence levels, whereas strains from different phylogenetic groups exhibited different predispositions to decreased sensitivity to tebuconazole and fludioxonil. In vitro adaptation experiments were conducted to assess: (1) the potential ability of M. nivale to acquire high-level resistance to these fungicides; (2) the relative adaptation efficiency to each fungicide; and (3) the impact of resistance acquisition on virulence. Our results showed that M. nivale strains could adapt to high concentrations of both fungicides with little or no effect on virulence. Adaptation to fludioxonil was significantly less effective than to tebuconazole. To get closer to understanding the mechanisms of fludioxonil adaptation in M. nivale, whole-genome sequencing was performed on a fludioxonil-adapted derivative and its parental fludioxonil-sensitive strain. Comparative genome analysis identified mutations potentially involved in the enhanced fludioxonil resistance, which are discussed within the framework of molecular resistance mechanisms.
Fungicide treatment is one of the most common methods for controlling fungal plant diseases. However, many phytopathogenic fungi develop resistance to fungicides. Addressing this agriculturally important issue requires comprehensive investigations into fungicide resistance. Our study aims to assess the degree and prevalence of resistance to carbendazim—one of the most widely used fungicides—in populations of Microdochium nivale, the causal agent of the deleterious plant disease pink snow mold; to explore possible relationships between carbendazim resistance and physiological and genetic traits; and to gain insight into the molecular basis of carbendazim resistance in this species. We showed that carbendazim resistance is widespread in the analyzed M. nivale populations, and that the application of carbendazim increases the proportion of resistant strains. Nevertheless, carbendazim-resistant strains are present at high relative abundance in populations that have never been exposed to fungicides. Carbendazim resistance in M. nivale is strongly associated with sequence variations in the β-tubulin gene, resulting in amino acid sequence variability that leads to differential affinity for carbendazim. Additionally, we propose a metabarcoding-based approach employing a genetic marker linked to a specific phenotypic trait to assess the ratio of genotypes with contrasting properties within a particular fungal species in environmental communities.
Fusarium head blight (FHB) is a major threat to cereal crops, causing yield losses and mycotoxin contamination. This study investigated Fusarium species associated with spikes and grains of cereals in the Volga region, focusing on species diversity, virulence, and mycotoxin production. F. sporotrichioides, F. avenaceum, and F. poae were the most prevalent species isolated from wheat, rye, barley, triticale, and stored grains in the Volga region. Individual strains of F. culmorum and F. graminearum were also identified. High intraspecific variability in virulence was observed for the first time within F. sporotrichioides and F. poae species, and highly virulent strains were identified for the first time within these species. Not only symptomatic but also asymptomatic (weakly expressed) infections caused by F. sporotrichioides were shown to be associated with the accumulation of high levels of T-2 toxin in the grains of infected plants. F. sporotrichioides strains were first demonstrated to exhibit intraspecific variability in zearalenone-producing potential. A F. graminearum strain possessing the nivalenol chemotype was first identified in Russia. The study highlights the diversity of the regional FHB pathocomplex and the risks it poses to grain safety.