Skip to main content
SpringerLink
Log in

Characterization of spontaneous mutants of Magnaporthe grisea expressing stable resistance to the Qo-inhibiting fungicide azoxystrobin

  • Research Article
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract.

The class of Qo-inhibiting fungicides (QoIs) act as respiration inhibitors by binding to the Qo center of cytochrome b. The longevity of these fungicides has been challenged by the selection of fungal sub-populations resisting high doses of QoI fungicides, with a G143A amino acid exchange in the cytochrome b target site identified as the most common cause of resistance. In contrast, the mechanism of alternative respiration, as another mechanism of fungal QoI resistance, has thus far not been affiliated with practical resistance. In the present study, azoxystrobin-resistant mutants of Magnaporthe grisea were generated and characterized. Emergence of these spontaneous mutants was facilitated when resting melanized mycelia were allowed to escape full inhibition by azoxystrobin. This escape was related to the intactness of alternative respiration, indicating that residual expression of this rescue mechanism was involved in the spontaneous emergence of target-site mutants. The two mutants characterized resisted high doses of the QoI, azoxystrobin, with resistance factors exceeding 1,000. Two different mutations of the cytochrome b gene were identified as exchanges of guanine, leading to a G143A or a G143S amino acid exchange. Resistance of both target-site mutants remained stable during four consecutive disease cycles in the absence of azoxystrobin. Several parameters tested to measure fitness penalties inherent to the mutational changes revealed that the G143A mutant was not compromised. In contrast, the conidia production of the G143S mutant was significantly lower under both saprophytic and pathogenic conditions of reproduction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1A–C.
Fig. 2A, B.

Similar content being viewed by others

References

  • Altschul SF, Madden TL, Schäffer AA, Zhang, J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    PubMed  Google Scholar 

  • Avila-Adame C, Köller W (2002) Disruption of the alternative oxidase gene in Magnaporthe grisea and its impact on host infection. Mol Plant-Microbe Interact 15:493–500

    CAS  Google Scholar 

  • Balhadère PV, Foster AJ, Talbot NJ (1999) Identification of pathogenicity mutants of the rice blast fungus Magnaporthe grisea by insertional mutagenesis. Mol Plant-Microbe Interact 12:129–142

    Google Scholar 

  • Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M, Parr-Dobrzanski B (2002) The strobilurin fungicides. Pest Manag Sci 58:649–662

    Article  CAS  PubMed  Google Scholar 

  • Bohr VA, Anson RM (1999) Mitochondrial DNA repair pathways. J Bioenerg Biomembr 31:391–398

    Article  CAS  PubMed  Google Scholar 

  • Crawford MS, Chumley FG, Weaver CG, Valent B (1986) Characterization of the heterokaryotic and vegetative diploid phases of Magnaporthe grisae. Genetics 114:1111–1129

    Google Scholar 

  • Csaikl UM, Bastian H, Brettschneider R, Gauch S, Meir A, Schauerte M, Scholz F, Sperisen C, Vornam B, Ziegenhagen B (1998) Comparative analysis of different DNA extraction protocols: a fast, universal maxi-preparation of high quality plant DNA for genetic evaluation and phylogenetic studies. Plant Mol Biol Rep 16:69–86

    Article  CAS  Google Scholar 

  • Dekker J (1995) Development of resistance to modern fungicides and strategies for its avoidance. In: Lyr H (ed) Modern selective fungicides. Fisher, Jena, pp 23–38

  • DeZwaan TM, Carroll AM, Valent B, Sweigard JA (1999) Magnapothe grisea Pth11p is a novel plasma membrane protein that mediates appressorium differentiation in response to inductive substrate cues. Plant Cell 11:2013–2030

    CAS  PubMed  Google Scholar 

  • Di Rago JP, Hermann-Le Denmant S, Paques F, Risler J, Netter P, Slonimski PP (1995) Genetic analysis of the folded structure of yeast mitochondrial cytochrome b by selection of intragenic second-site revertants. J Mol Biol 248:804–811

    Article  PubMed  Google Scholar 

  • Heaney SP, Hall AA, Davies SA, Olaya G (2000) Resistance to fungicides in the QoI-STAR cross-resistance group: current perspectives. Proc Br Crop Prot Conf Pests Dis 2000:755–764

    Google Scholar 

  • Ishii H, Fraaije BA, Sugiyama T, Noguchi K, Nishimura K, Takeda T, Amano T, Hollomon DW (2001) Occurrence and molecular characterization of strobilurin resistance in cucumber powdery mildew and downy mildew. Phytopathology 91:1166–1171

    CAS  Google Scholar 

  • Joseph-Horne T, Hollomon DW (2000) Functional diversity within the mitochondrial electron transport chain of plant pathogenic fungi. Pest Manag Sci 56:24–30

    Article  CAS  Google Scholar 

  • Joseph-Horne T, Hollomon DW, Wood PM (2001) Fungal respiration: a fusion of standard and alternative components. Biochim Biophys Acta 1504:179–195

    CAS  PubMed  Google Scholar 

  • Köller W (1991) Fungicide resistance in crop protection. In: Pimentel D (ed) CRC handbook of pest management in agriculture. CRC Press, Boca Raton, Fla., pp 679–720

  • Köller W (1999) Chemical approaches to managing plant pathogens. In: Ruberson JR (ed) Handbook of pest management. Dekker, New York, pp 337–376

  • Köller W (2001) Fungicide resistance. In: Maloy OC, Murray TD (eds) Encyclopedia of plant pathology. Wiley, New York, pp 483–488

  • Köller W, Avila-Adame C, Olaya G, Zheng D (2001) Resistance to strobilurin fungicides. In: Clark JM, Yamaguchi I (eds) Agrochemical resistance extent, mechanism, and detection. American Chemical Society, Washington, D.C., pp 215–229

  • Milgroom MG, Levin SA, Fry WE (1989) Population genetics theory and fungicide resistance. In: Leonard KJ, Fry WE (eds) Plant disease epidemiology: genetics, resistance and management, vol 2. McGraw-Hill, New York, pp 340–367

    Google Scholar 

  • Olaya G, Köller W (1999) Baseline sensitivities of Venturia inaequalis populations to the strobilurin fungicide kresoxim-methyl. Plant Dis 83:274–278

    CAS  Google Scholar 

  • Olaya G, Zheng D, Köller W (1998) Differential responses of germinating Venturia inaequalis conidia to kresoxim-methyl. Pestic Sci 54:230–236

    Article  CAS  Google Scholar 

  • Rosenberg SM (2001) Evolving responsively: adaptive mutation. Nat Rev Genet 2:504–515

    CAS  PubMed  Google Scholar 

  • Sauter H, Steglich W, Anke T (1999) Strobilurins: evolution of a new class of active substances. Angew Chem Int Ed 38:1328–1349

    Article  Google Scholar 

  • Sawer DE, Van Houten B (1999) Repair of DNA damage in mitochondria. Mutat Res 434:161–176

    CAS  PubMed  Google Scholar 

  • Sierotzki H, Parisi S, Steinfeld U, Tenzer I, Poirey S, Gisi U (2000a) Mode of resistance to respiration inhibitors at the cytochrome bc1 enzyme complex of Mycosphaerella fijiensis field isolates. Pest Manag Sci 56: 833–841

    Article  CAS  Google Scholar 

  • Sierotzki H, Wullschleger J, Gisi U (2000b) Point mutation in cytochrome b gene conferring resistance to strobilurin fungicides in Erysiphe graminis f. sp. tritici field isolates. Pestic Biochem Physiol 68:107–112

    Article  CAS  Google Scholar 

  • Singh KK, Sigala B, Sikder HA, Schwimmer C. (2001) Inactivation of Saccharomyces cerevisiae OGG1 DNA repair gene leads to an increased frequency of mitochondrial mutants. Nucleic Acids Res 29:1381–1388

    CAS  PubMed  Google Scholar 

  • Skylakakis G (1987) Changes in the composition of pathogen populations caused by resistance to fungicides. In: Wolfe MS, Caten CE (eds) Populations of plant pathogens: their dynamics and genetics. Blackwell, Oxford, pp 227–237

    Google Scholar 

  • Wolfe MS (1982) Dynamics of the pathogen population in relation to fungicide resistance. In: Dekker J, Georgopoulos SG (eds) Fungicide resistance in crop protection. Centre for agricultural publishing and documentation, Wageningen, pp 139–148

  • Ypema HL, Gold RE (1999) Kresoxim-methyl. modification of a naturally occurring compound to produce a new fungicide. Plant Dis 83:4–19

    CAS  Google Scholar 

  • Yukioka H, Inagaki S, Tanaka R, Katoh K, Miki N, Mizutani A, Masuko M (1998) Transcriptional activation of the alternative oxidase gene of the fungus Magnaporthe grisea by a respiratory-inhibiting fungicide and hydrogen peroxide. Biochim Biophys Acta 1442:161–169

    Article  CAS  PubMed  Google Scholar 

  • Zheng D, Olaya G, Köller W (2000) Characterization of a laboratory mutant of Venturia inaequalis resistant to the strobilurin-related fungicide kresoxim-methyl. Curr Genet 38:148–155

    Article  CAS  PubMed  Google Scholar 

  • Ziogas BN, Baldwin BC, Young JE (1997) Alternative respiration: a biochemical mechanism of resistance to azoxystrobin (ICIA 5504) in Septoria tritici. Pestic Sci 50: 28–34

    Article  CAS  Google Scholar 

Download references

Acknowledgements.

This work was supported, in part, by financial support to C.A.-A. provided by the Fulbright–Garcia Robles foundation, the Institute of International Education, CONACYT–Mexico and the Colegio de Postgraduados–Mexico.

Author information

Author notes

  1. Cruz Avila-Adame

    Present address: Departamento de Fitopatologia, Colegio de Postgraduados Campus Tabasco, H. Cárdenas, Tabasco, CP 86500, México

Authors and Affiliations

  1. Department of Plant Pathology, Cornell University, New York State Agricultural Experiment Station, Geneva, NY 14456, USA

    Cruz Avila-Adame & Wolfram Köller

Authors
  1. Cruz Avila-Adame
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfram Köller
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wolfram Köller.

Additional information

Communicated by U. Kück

Rights and permissions

Reprints and permissions

About this article

Cite this article

Avila-Adame, C., Köller, W. Characterization of spontaneous mutants of Magnaporthe grisea expressing stable resistance to the Qo-inhibiting fungicide azoxystrobin. Curr Genet 42, 332–338 (2003). https://doi.org/10.1007/s00294-002-0356-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00294-002-0356-1

Keywords.

Search

Navigation