ABSTRACT
Wheat stem rust, a devastating disease of wheat and barley caused by the fungal pathogen Puccinia graminis f. sp. tritici, was largely eradicated in Western Europe during the mid-to-late twentieth century. However, isolated outbreaks have occurred in recent years. Here we investigate whether a lack of resistance in modern European varieties, increased presence of its alternate host barberry and changes in climatic conditions could be facilitating its resurgence. We report the first wheat stem rust occurrence in the United Kingdom in nearly 60 years, with only 20% of UK wheat varieties resistant to this strain. Climate changes over the past 25 years also suggest increasingly conducive conditions for infection. Furthermore, we document the first occurrence in decades of P. graminis on barberry in the UK . Our data illustrate that wheat stem rust does occur in the UK and, when climatic conditions are conducive, could severely harm wheat and barley production.
ABSTRACT
BACKGROUND: The major facilitator superfamily (MFS) drug transporter MgMfs1 of the wheat pathogen Mycosphaerella graminicola (Fuckel) J Schroeter is a potent multidrug transporter with high capacity to transport strobilurin fungicides in vitro. The data presented in this paper indicate that, in addition to the predominant cause of strobilurin resistance, cytochrome b G143A subsititution, MgMfs1 can play a role in sensitivity of field strains of this pathogen to trifloxystrobin. RESULTS: In a major part of field strains of M. graminicola (collected in the Netherlands in 2004) containing the cytochrome b G143A substitution, the basal level of expression of MgMfs1 was elevated as compared with sensitive strains lacking the G143A substitution. Induction of MgMfs1 expression in wild-type isolates upon treatment with trifloxystrobin at sublethal concentrations proceeded rapidly. Furthermore, in disease control experiments on wheat seedlings, disruption mutants of MgMfs1 displayed an increased sensitivity to trifloxystrobin. CONCLUSION: It is concluded that the drug transporter MgMfs1 is a determinant of strobilurin sensitivity of field strains of M. graminicola.
Subject(s)
Acetates/pharmacology , Ascomycota/drug effects , Ascomycota/metabolism , Fungal Proteins/metabolism , Fungicides, Industrial/pharmacology , Imines/pharmacology , Membrane Transport Proteins/metabolism , Plant Diseases/microbiology , Triticum/microbiology , Acetates/metabolism , Amino Acid Substitution , Ascomycota/genetics , Ascomycota/isolation & purification , Cytochrome b Group/genetics , Cytochrome b Group/metabolism , Fungal Proteins/genetics , Fungicides, Industrial/metabolism , Imines/metabolism , Membrane Transport Proteins/genetics , Methacrylates/metabolism , Methacrylates/pharmacology , Microbial Sensitivity Tests , Mutation , Strobilurins , Triticum/drug effectsABSTRACT
Medical drugs known to modulate the activity of human ATP-binding cassette (ABC) transporter proteins (modulators) were tested for the ability to potentiate the activity of the azole fungicide cyproconazole against in vitro growth of Mycosphaerella graminicola and to control disease development due to this pathogen on wheat seedlings. In vitro modulation of cyproconazole activity could be demonstrated in paper disk bioassays. Some of the active modulators (amitriptyline, flavanone, and phenothiazines) increased the accumulation of cyproconazole in M. graminicola, suggesting that they reversed cyproconazole efflux. However, synergism between cyproconazole and modulators against M. graminicola on wheat seedlings could not be shown. Despite their low in vitro toxicity to M. graminicola, some modulators (amitriptyline, loperamide, and promazine) did show significant intrinsic disease control activity in preventive and curative foliar spray tests with wheat seedlings. The results suggest that these compounds have indirect disease control activity based on modulation of fungal ABC transporters essential for virulence and constitute a new class of disease control agents.
Subject(s)
ATP-Binding Cassette Transporters/metabolism , Antifungal Agents/pharmacology , Ascomycota/drug effects , Gene Expression Regulation, Fungal , Seedlings/microbiology , Triticum/microbiology , ATP-Binding Cassette Transporters/genetics , Adrenergic Uptake Inhibitors/pharmacology , Amitriptyline/pharmacology , Antidiarrheals/pharmacology , Ascomycota/growth & development , Dopamine Antagonists/pharmacology , Drug Synergism , Fungal Proteins , Loperamide/pharmacology , Microbial Sensitivity Tests , Plant Diseases/microbiology , Promazine/pharmacology , Triazoles/pharmacologyABSTRACT
The ABC transporter-encoding gene MgAtr7 from the wheat pathogen Mycosphaerella graminicola was cloned based upon its high homology to ABC transporters involved in azole-fungicide sensitivity. Genomic and cDNA sequences indicated that the N-terminus of this ABC transporter contains a motif characteristic for a dityrosine/pyoverdine biosynthesis protein. This makes MgAtr7 the first member of a new class of fungal ABC transporters harboring both a transporter and a biosynthetic moiety. A homologue of MgAtr7 containing the same biosynthetic moiety was only found in the Fusarium graminearum genome and not in any other fungal genome examined so far. The gene structure of both orthologous transporters is highly conserved and the genomic area surrounding the ABC transporter exhibits micro-synteny between M. graminicola and F. graminearum. Functional analyses revealed that MgAtr7 is neither required for virulence nor involved in fungicide sensitivity but indicated a role in maintenance of iron homeostasis.
Subject(s)
ATP-Binding Cassette Transporters/physiology , Ascomycota/physiology , Homeostasis/physiology , Iron/metabolism , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Ascomycota/genetics , Drug Resistance, Fungal/genetics , Drug Resistance, Fungal/physiology , Fungal Proteins/metabolism , Gene Expression Regulation, Fungal , Plant Diseases/microbiologyABSTRACT
MgMfs1, a major facilitator superfamily (MFS) gene from the wheat pathogenic fungus Mycosphaerella graminicola, was identified in expressed sequence tag (EST) libraries. The encoded protein has high homology to members of the drug:H(+) antiporter efflux family of MFS transporters with 14 predicted transmembrane spanners (DHA14), implicated in mycotoxin secretion and multidrug resistance. Heterologous expression of MgMfs1 in a hypersensitive Saccharomyces cerevisiae strain resulted in a strong decrease in sensitivity of this organism to a broad range of unrelated synthetic and natural toxic compounds. The sensitivity of MgMfs1 disruption mutants of M. graminicola to most of these compounds was similar when compared to the wild-type but the sensitivity to strobilurin fungicides and the mycotoxin cercosporin was increased. Virulence of the disruption mutants on wheat seedlings was not affected. The results indicate that MgMfs1 is a true multidrug transporter that can function as a determinant of pathogen sensitivity and resistance to fungal toxins and fungicides.
Subject(s)
Antifungal Agents/pharmacology , Ascomycota/drug effects , Fungal Proteins/physiology , Fungicides, Industrial/pharmacology , Membrane Transport Proteins/physiology , Triticum/microbiology , Ascomycota/genetics , Ascomycota/metabolism , Drug Resistance, Multiple, Fungal/genetics , Expressed Sequence Tags , Fungal Proteins/genetics , Fungal Proteins/metabolism , Genes, Fungal/genetics , Ketoconazole/pharmacology , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Miconazole/pharmacology , Microbial Sensitivity Tests , Models, Biological , Mutation/genetics , Mutation/physiology , Perylene/analogs & derivatives , Perylene/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/genetics , Triazoles/pharmacologyABSTRACT
Chitinase specific activity was measured spectrophotometrically in wheat leaf tissues during the compatible and incompatible interactions with Puccinia striiformis f. sp. tritici, the causal agent of yellow rust disease. The wheat cultivar, Federation* 4/Kavkaz, was inoculated with virulent (134E134A+) or avirulent (4EOA+) races of P. striiformis f. sp. tritici in the first leaf stage. The results showed that chitinase activity pattern was similar in both compatible and incompatible interactions up to 72 hrs after inoculation. However, the specific activity increased rapidly in the incompatible reaction thereafter. In susceptible reaction, chitinase activity gradually declined after 72 hrs post-inoculation reaching a level similar to that in the control plants two weeks after inoculation. Chitinase specific activity in resistance response was at least three times greater than that in the susceptible reaction two weeks following the inoculation. Electrophoresis of native polyacrylamide gel impregnated with 0.1% (w/v) glycol chitin as the substrate revealed the presence of eight chitinase isoforms with relative electrophoretic mobility (Rm) values ranging from 0.11 to 0.64 in the resolving gel.