Abstract
Mutants of Botrytis cinerea with moderate and high resistance to pyraclostrobin, a Qo inhibitor of mitochondrial electron transport at the cytochrome bc 1 complex, were isolated at a high mutation frequency, after nitrosoguanidine mutagenesis and selection on medium containing pyraclostrobin and salicylhydroxamate (SHAM), a specific inhibitor of cyanide-resistant (alternative) respiration. Oxygen uptake in whole cells was strongly inhibited in the wild-type strain by pyraclostrobin and SHAM, but not in the mutant isolates. Cross-resistance studies with other Qo and Qi inhibitors (QoIs and QiIs) of cytochrome bc 1 complex of mitochondrial respiration showed that the mutation(s) for resistance to pyraclostrobin also reduced the sensitivity of mutant strains to other QoIs as azoxystrobin, fluoxastrobin, trifloxystrobin and picoxystrobin, but not to famoxadone and to the QiIs cyazofamid and antimycin-A. An increased sensitivity of pyraclostrobin-resistant strains to the carboxamide boscalid, an inhibitor of complex II, and to the anilinopyrimidine cyprodinil, a methionine biosynthesis inhibitor, was observed. Moreover, no effect of pyraclostrobin resistance mutation(s) on fungitoxicity of the hydroxyanilide fenhexamid, the phenylpyrrole fludioxonil, the benzimidazole benomyl, and to the phenylpyridinamine fluazinam, which affect other cellular pathways, was observed. Study of fitness parameters in the wild-type and pyraclostrobin-resistant mutants of B. cinerea showed that most mutants had a significant reduction in the sporulation, conidial germination and sclerotia production. Experiments on the stability of the pyraclostrobin-resistant phenotype showed a reduction of resistance, mainly in moderate resistant strains, when the mutants were grown on inhibitor-free medium. However, a rapid recovery of the resistance level was observed after the mutants were returned to a selective medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of high resistant mutants was higher than the moderate ones. Pathogenicity tests on cucumber seedlings showed that all mutant strains tested exhibited an infection ability similar with the wild-type parent strain. Preventive applications of the commercial product of F-500 25EC (pyraclostrobin) were effective against lesion development on cotyledons by the wild-type, but ineffective, even at high concentrations, against disease caused by the pyraclostrobin-resistant isolates. Boscalid (F-510 50WG) was found equally effective against the disease caused by the wild-type or pyraclostrobin-resistant mutants. This is the first report indicating the appearance of B. cinerea strains resistant to QoI fungicides by the biochemical mechanism of site modification and the risk for field resistance.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Ammermann E, Lorenz G, Schelberger K, Mueller B, Kirstgen R and Sauter H (2000) BAS 500 F – the new broad-spectrum strobilurin fungicide. In: Proceedings of Brighton Crop Protection Conference, Pests and Diseases, Vol. 2 (pp. 541–548) British Crop Protection Council, Farnham, Surrey, UK
Avila-Adame C and Köller W (2003). Characterization of spontaneous mutants of Magnaporthe grisea expressing stable resistance to the Qo-inhibiting fungicide azoxystrobin. Current Genetics 42: 332–338
Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M and Parr-Dobrzanski B (2002). The strobilurin fungicides. Pest Management Science 58: 649–662
Brent KJ and Hollomon DW (1998) Fungicide resistance: The assessment of risk. In: FRAC Monograph No 2. (pp. 1–48) Global Crop Protection Federation, Brussels, Belgium
Chapeland F, Fritz R, Lanen C, Gredt M and Leroux P (1999). Inheritance and mechanisms of resistance to anilinopyrimidine fungicides in Botrytis cinerea (Botryotinia fuckeliana). Pesticide Biochemistry and Physiology 64: 85–100
Chin KM, Chavaillaz D, Kaesbohrer M, Staub T and Felsenstein FG (2001a). Characterizing resistance risk of Erysiphe graminis f.sp. tritici to strobilurins. Crop Protection 20: 87–96
Chin KM, Wirz M and Laird D (2001b). Sensitivity of Mycosphaerella fijiensis from banana to trifloxystrobin. Plant Disease 85: 1264–1270
Collina M, Landi L, Guerrini P, Branzanti MB and Brunelli A (2004) QoI resistance of Plasmopara viticola in Italy: Biological and quantitative real-time PCR approaches. In: 14th International Reinhardsbrunn Symposium, Modern Fungicides and Antifungal Compounds, Friedrichroda, Germany
Di Rago JP and Colson AM (1989). Molecular basis for resistance to myxothiazol, mucidin (strobilurin A) and stigmatellin. The Journal of Biological Chemistry 264: 14543–14548
Faretra F and Pollastro S (1993). Isolation, characterization and genetic analysis of laboratory mutants of Botryotinia fuckeliana resistant to the phenylpyrrole fungicide CGA 173506. Mycological Research 97: 620–624
Farman ML (2001). The molecular basis of field resistance to QoI fungicides in Pyricularia grisea. Phytopathology (Abstr.) 91: S110
Heaney SP, Hall AA, Davies SA and Olaya G (2000) Resistance to fungicides in the QoI-STAR cross-resistance group: Current perspectives. In: Proceedings of Brighton Crop Protection Conference, Pests and Diseases, Vol. 2 (pp. 755–762) British Crop Protection Council, Farnham, Surrey, UK
Ishii H, Fraaije BA, Sugiyama T, Noguchi K, Nishimura K, Takeda T, Amano T and Hollomon DW (2001). Occurrence and molecular characterization of strobilurin resistance in cucumber powdery mildew and downy mildew. Phytopathology 91: 1166–1171
Joseph-Horne T, Wood PM, Heppner C and Hollomon DW (1999). Involvement of the alternative oxidase in cellular energy production in the wheat ‘Take-All’ fungus, Gaeumannomyces graminis var. tritici. Pesticide Science 55: 367–370
Kim YS, Dixon EW, Vincelli P and Farman ML (2003). Field resistance to strobilurin (QoI) fungicides in Pyricularia grisea caused by mutations in the mitochondrial cytochrome b gene. Phytopathology 93: 891–900
Leroux P, Chapeland F, Desbrosses D and Gredt M (1999). Patterns of cross-resistance to fungicides in Botryotinia fuckeliana (Botrytis cinerea) isolates from French vineyards. Crop Protection 18: 687–697
Ma Z, Felts D and Michailides TJ (2003). Resistance to azoxystrobin in Alternaria isolates from pistachio in California. Pesticide Biochemistry and Physiology 77: 66–74
Olaya G and Köller W (1999a). Diversity of kresoxim-methyl sensitivities in baseline populations of Venturia inaequalis. Pesticide Science 55: 1083–1088
Olaya G and Köller W (1999b). Baseline sensitivities of Venturia inaequalis populations to the strobilurin fungicide kresoxim-methyl. Plant Disease 83: 274–278
Pasche JS, Piche LM and Gudmestad NC (2005). Effect of the F129L mutation in Alternaria solani on fungicides affecting mitochondrial respiration. Plant Disease 89: 269–278
Sierotzki H, Wullschleger J and Gisi U (2000a). Point-mutation in cytochrome b gene confering resistance to strobilurin fungicides in Erysiphe graminis f.sp. tritici field isolates. Pesticide Biochemistry and Physiology 68: 107–112
Sierotzki H, Parisi S, Steinfeld U, Tenzer I, Poirey S and Gisi U (2000b). Mode of resistance to respiration inhibitors at the cytochrome bc 1 enzyme complex of Mycosphaerella fijiensis field isolates. Pest Management Science 56: 833–841
Sierotzki H, Kraus N, Assemat P, Stanger C, Cleere S, Windass J and Gisi U (2004) Evolution of resistance to QoI fungicides in Plasmopara viticola in Europe. In: 14th International Reinhardsbrunn Symposium, Modern Fungicides and Antifungal Compounds, Friedrichroda, Germany
Steinfeld U, Sierotzki H, Parisi S, Poirey S and Gisi U (2001). Sensitivity of mitochondrial respiration to different inhibitors in Venturia inaequalis. Pest Management Science 57: 787–796
Tamura H, Mizutani A, Yukioka H, Miki N, Ohba K and Masuko M (1999). Effect of the methoxyiminoacetamide fungicide, SSF129, on respiratory activity of Botrytis cinerea. Pesticide Science 55: 681–686
Vincelli P and Dixon E (2002). Resistance to QoI (strobilurin-like) fungicides in isolates of Pyricularia grisea from perennial ryegrass. Plant Disease 86: 235–240
Yukioka H, Tanaka R, Inagaki S, Katoh K, Miki N, Mizutani A, Masuko M and Kunoh H (1997). Mutants of the phytopathogenic fungus Magnaporthe grisea deficient in alternative, cyanide-resistant respiration. Fungal Genetics and Biology 22: 221–228
Zheng D and Köller W (1997). Characterization of the mitochondrial cytochrome b gene from Venturia inaequalis. Current Genetics 32: 361–366
Zheng D, Olaya G and Köller W (2000). Characterization of laboratory mutants of Venturia inaequalis resistant to the strobilurin-related fungicide kresoxim-methyl. Current Genetics 38: 148–155
Ziogas BN and Girgis SM (1993). Cross-resistance relationships between benzimidazole fungicides and diethofencarb in Botrytis cinerea and their genetical basis in Ustilago maydis. Pesticide Science 39: 199–205
Ziogas BN and Kalamarakis AE (2001). Phenylpyrrole fungicides: Mitotic instability in Aspergillus nidulans and resistance in Botrytis cinerea. Journal of Phytopathology 149: 301–308
Ziogas BN, Baldwin BC and Young JE (1997). Alternative respiration: A biochemical mechanism of resistance to azoxystrobin (ICIA 5504) in Septoria tritici. Pesticide Science 50: 28–34
Ziogas BN, Markoglou AN and Tzima A (2002). A non-Mendelian inheritance of resistance to strobilurin fungicides in Ustilago maydis. Pest Management Science 58: 908–916
Ziogas BN, Markoglou AN and Malandrakis AA (2003). Studies on the inherent resistance risk to fenhexamid in Botrytis cinerea. European Journal of Plant Pathology 109: 311–317
Ziogas BN, Markoglou AN and Spyropoulou V (2005). Effect of phenylpyrrole-resistance mutations on ecological fitness of Botrytis cinerea and their genetical basis in Ustilago maydis. European Journal of Plant Pathology 113: 83–100
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Markoglou, A.N., Malandrakis, A.A., Vitoratos, A.G. et al. Characterization of Laboratory Mutants of Botrytis cinerea Resistant to QoI Fungicides. Eur J Plant Pathol 115, 149–162 (2006). https://doi.org/10.1007/s10658-006-0008-2
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10658-006-0008-2