ABSTRACT
Plants commandeer a pathogen's virulence factor to bolster immunity.
Subject(s)
Fusarium , Phaseolus , Plant Diseases , Plant Immunity , Plant Proteins , Virulence Factors , Plant Diseases/immunology , Virulence Factors/metabolism , Fusarium/immunology , Fusarium/pathogenicity , Phaseolus/immunology , Phaseolus/microbiology , Plant Proteins/metabolismABSTRACT
White grain disorder is a recently emerged wheat disease in Australia, caused by Eutiarosporella darliae, E. pseudodarliae, and E. tritici-australis. The disease cycle of these pathogens and the molecular basis of their interaction with wheat are poorly understood. To address this knowledge gap, we undertook a comparative genomics analysis focused on the secondary metabolite gene repertoire among these three species. This analysis revealed a diverse array of secondary metabolite gene clusters in these pathogens, including modular polyketide synthase genes. These genes have only been previously associated with bacteria and this is the first report of such genes in fungi. Subsequent phylogenetic analyses provided strong evidence that the modular PKS genes were horizontally acquired from a bacterial or a protist species. We also uncovered a secondary metabolite gene cluster with three polyketide/nonribosomal peptide synthase genes (Hybrid-1, -2, and -3) in E. darliae and E. pseudodarliae. In contrast, only remnant and partial genes homologous to this cluster were identified in E. tritici-australis, suggesting loss of this cluster. Homologues of Hybrid-2 in other fungi have been proposed to facilitate disease in woody plants, suggesting a possible alternative host range for E. darliae and E. pseudodarliae. Subsequent assays confirmed that E. darliae and E. pseudodarliae were both pathogenic on woody plants, but E. tritici-australis was not, implicating woody plants as potential host reservoirs for the fungi. Combined, these data have advanced our understanding of the lifestyle and potential host-range of these recently emerged wheat pathogens and shed new light on fungal secondary metabolism.
Subject(s)
Ascomycota/genetics , Biological Evolution , Polyketide Synthases/genetics , Triticum/microbiology , Ascomycota/metabolism , Ascomycota/pathogenicity , Gene Duplication , Gene Transfer, Horizontal , Genome, Fungal , Plant Diseases , Secondary MetabolismABSTRACT
White grain disorder (WGD) is a recently emerged wheat disease in Australia caused by three Botryosphaeriaceae fungi, from the genus Eutiarosporella. These species are E. tritici-australis, E. darliae, and E. pseudodarliae. Characterisation of the mating type genes for the WGD-species show that the genome sequence of a single E. darliae and E. pseudodarliae isolate both harbour MAT1-2-1 and MAT1-1-1, which suggests that these species are homothallic. However, unlike most other characterised mating-type loci from other homothallic Dothideomycetes, these species' MAT1-1-1 are located at a separate locus, inserted within the coding region of another gene. The sequenced strain of E. tritici-australis analysed did not harbour MAT1-1-1. Including the sequenced strain, we screened the mating type genes present in 16 E. tritici-australis individuals isolated from infected grain from fields in South Australia. Of these 16, 11 harbour MAT1-1-1 and the other five harbour MAT1-2-1. The genome of a MAT1-1-1 harbouring isolate was re-sequenced, which demonstrated that MAT1-1-1 was present at the MAT locus. We examined non-coding DNA surrounding the MAT1-1-1 gene in E. pseudodarliae and observed fragments of the MAT locus both up and downstream. These fragments and their orientation around MAT1-1-1 is similar to characterised heterothallic Botryosphaeriaceae. Based on these gene arrangements, we conclude that the new MAT1-1-1 containing locus likely originated from a cryptic DNA integration event between two heterothallic individuals. We hypothesise that this integration event led to the formation of a homothallic lineage, which is the common ancestor of E. darliae and E. pseudodarliae.
Subject(s)
Ascomycota/physiology , Genes, Mating Type, Fungal , Genetic Speciation , Triticum/microbiology , Ascomycota/genetics , Ascomycota/growth & development , Fertility/geneticsABSTRACT
In this article, we describe the presence of genes encoding close homologues of an endogenous plant peptide, rapid alkalinization factor (RALF), within the genomes of 26 species of phytopathogenic fungi. Members of the RALF family are key growth factors in plants, and the sequence of the RALF active region is well conserved between plant and fungal proteins. RALF1-like sequences were observed in most cases; however, RALF27-like sequences were present in the Sphaerulina musiva and Septoria populicola genomes. These two species are pathogens of poplar and, interestingly, the closest relative to their respective RALF genes is a poplar RALF27-like sequence. RALF peptides control cellular expansion during plant development, but were originally defined on the basis of their ability to induce rapid alkalinization in tobacco cell cultures. To test whether the fungal RALF peptides were biologically active in plants, we synthesized RALF peptides corresponding to those encoded by two sequenced genomes of the tomato pathogen Fusarium oxysporum f. sp. lycopersici. One of these peptides inhibited the growth of tomato seedlings and elicited responses in tomato and Nicotiana benthamiana typical of endogenous plant RALF peptides (reactive oxygen species burst, induced alkalinization and mitogen-activated protein kinase activation). Gene expression analysis confirmed that a RALF-encoding gene in F. oxysporum f. sp. lycopersici was expressed during infection on tomato. However, a subsequent reverse genetics approach revealed that the RALF peptide was not required by F. oxysporum f. sp. lycopersici for infection on tomato roots. This study has demonstrated the presence of functionally active RALF peptides encoded within phytopathogens that harbour an as yet undetermined role in plant-pathogen interactions.
Subject(s)
Fungal Proteins/metabolism , Fusarium/metabolism , Fusarium/pathogenicity , Peptide Hormones/metabolism , Plant Proteins/metabolism , Fungal Proteins/genetics , Fusarium/genetics , Solanum lycopersicum/genetics , Solanum lycopersicum/metabolism , Solanum lycopersicum/microbiology , Peptide Hormones/genetics , Plant Proteins/genetics , Seedlings/genetics , Seedlings/metabolism , Seedlings/microbiology , Nicotiana/genetics , Nicotiana/metabolism , Nicotiana/microbiologyABSTRACT
Phytopathogens are a global threat to plant agriculture and biodiversity. The genomics era has lead to an exponential rise in comparative gene and genome studies of both economically significant and insignificant microorganisms. In this review we highlight some recent comparisons and discuss how they identify shared genes or genomic regions associated with host virulence. The two major mechanisms of rapid genome adaptation - horizontal gene transfer and hybridisation - are reviewed and we consider how intra-specific pan-genome sequences encode alternative host specificity. We also discuss the power that access to expansive gene databases provides in aiding the study of phytopathogen emergence. These databases can rapidly enable the identification of an unknown pathogen and its origin, as well as genomic adaptations required for emergence.