Screening for resistance to ripe rot caused by Colletotrichum acutatum in grape germplasm

We screened 235 Vitis and six Muscadinia grapevine cultivars and selections conserved at the National Institute of Fruit Tree Science in Japan for resistance to grape ripe rot, caused by Colletotrichum acutatum Simmonds ex Simmonds. This fungus is insensitive to fungicides such as benomyl, diethofencarb, and iminoctadine-triacetate. We evaluated the disease resistance of nearly ripe berries from each cultivar and selection by artificial inoculation with C. acutatum. Analysis of variance of 20 cultivars and selections indicated that the genotype had a significant effect but that the year had no significant effect on the percentage of diseased berries. Genetic variance explained 85 % of total variance. Each cultivar or selection was classified into one of the following four classes based on its level of resistance to ripe rot: 50 highly resistant (≤ 20 % affected), 37 resistant (21- 40 %), 48 susceptible (41- 60 %), and 106 highly susceptible (≥ 61 %). Of the highly resistant cultivars and selections, we consider a diploid named 676-64 to be promising material for ripe rot resistant table grape breeding.

Transcriptome profiling on the response of Mycosphaerella graminicola isolates to an azole fungicide using cDNA arrays

Resistance to azole antifungals is a major problem in the control of diseases caused by fungal pathogens of both humans and plants. Potential for the development of azole resistance in the wheat leaf blotch pathogen Mycosphaerella graminicola, the causal agent of the most economically significant foliar disease of wheat in north-western Europe, is now of particular concern after the recent emergence of widespread resistance to quinone outside inhibitor fungicides. Using a cDNA microarray representing around 25% of the genome, we have profiled the transcriptional response of M. graminicola to epoxiconazole, currently the most widely used azole fungicide on cereal crops. By comparing the transcription profiles of two M. graminicola isolates with contrasting sensitivities to epoxiconazole we show qualitative and quantitative differences in differentially expressed genes, including those involved in ergosterol biosynthesis, mitochondrial respiration and transport mechanisms. This represents the first study investigating the response of a plant pathogenic fungus to a fungicide using cDNA microarray technology

Fungicide-Driven Evolution and Molecular Basis of Multidrug Resistance in Field Populations of the Grey Mould Fungus Botrytis cinerea

The grey mould fungus Botrytis cinerea causes losses of commercially important fruits, vegetables and ornamentals worldwide. Fungicide treatments are effective for disease control, but bear the risk of resistance development. The major resistance mechanism in fungi is target protein modification resulting in reduced drug binding. Multiple drug resistance (MDR) caused by increased efflux activity is common in human pathogenic microbes, but rarely described for plant pathogens. Annual monitoring for fungicide resistance in field isolates from fungicide-treated vineyards in France and Germany revealed a rapidly increasing appearance of B. cinerea field populations with three distinct MDR phenotypes. All MDR strains showed increased fungicide efflux activity and overexpression of efflux transporter genes. Similar to clinical MDR isolates of Candida yeasts that are due to transcription factor mutations, all MDR1 strains were shown to harbor activating mutations in a transcription factor (Mrr1) that controls the gene encoding ABC transporter AtrB. MDR2 strains had undergone a unique rearrangement in the promoter region of the major facilitator superfamily transporter gene mfsM2, induced by insertion of a retrotransposon-derived sequence. MDR2 strains carrying the same rearranged mfsM2 allele have probably migrated from French to German wine-growing regions. The roles of atrB, mrr1 and mfsM2 were proven by the phenotypes of knock-out and overexpression mutants. As confirmed by sexual crosses, combinations of mrr1 and mfsM2 mutations lead to MDR3 strains with higher broad-spectrum resistance. An MDR3 strain was shown in field experiments to be selected against sensitive strains by fungicide treatments. Our data document for the first time the rising prevalence, spread and molecular basis of MDR populations in a major plant pathogen in agricultural environments. These populations will increase the risk of grey mould rot and hamper the effectiveness of current strategies for fungicide resistance management