Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE (Open Access)

Fungicide sensitivity and resistance in the blackleg fungus, Leptosphaeria maculans, across canola growing regions in Australia

A. P. Van de Wouw https://orcid.org/0000-0001-5147-0393 A * , J. L. Scanlan A , S. J. Marcroft B , A. J. Smith B , E. M. Sheedy B , N. W. Perndt B , C. E. Harrison B , L. M. Forsyth C and A. Idnurm A
+ Author Affiliations
- Author Affiliations

A School of BioSciences, The University of Melbourne, Parkville, Vic. 3010, Australia.

B Marcroft Grains Pathology, Grains Innovation Park, Horsham, Vic. 3400, Australia.

C Syngenta Crop Protection, Macquarie Park, NSW 2113, Australia.

* Correspondence to: apvdw2@unimelb.edu.au

Handling Editor: Christian Huyghe

Crop & Pasture Science 72(12) 994-1007 https://doi.org/10.1071/CP21369
Submitted: 27 May 2021  Accepted: 20 September 2021   Published: 29 November 2021

© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)

Abstract

Fungicide use has become a fundamental part of many crop protection systems around the world, including to control blackleg disease on canola (Brassica napus L.). In Australia, most canola growers routinely apply at least one fungicide, and potentially multiple fungicides with different modes of action, in a single growing season. There is evidence for the emergence of fungicide resistance in Leptosphaeria maculans, the causal agent of blackleg disease, to the demethylation inhibitor (DMI) class of fungicides in Australia. However, it is not known whether resistance exists towards other chemical classes such as the succinate dehydrogenase inhibitors (SDHI). In this work, 397 samples were screened for resistance towards seven fungicide treatments in stubble-borne L. maculans populations collected from eight canola-growing agro-ecological regions of Australia from 2018 to 2020, a time frame that bridges the introduction of new chemicals for blackleg control. We confirmed that DMI resistance in L. maculans is pervasive across all of the sampled canola-growing regions, with 15% of fungal populations displaying high levels (resistance scores >0.5) of resistance towards the DMI fungicides. Although resistance to newly introduced SDHI fungicides was low, we found evidence of positive cross-resistance between established DMI-only fungicides and a newly introduced combined DMI and quinone outside inhibitor fungicide, suggesting that the efficacy of the latter may be limited by widespread DMI resistance. Proactive surveillance, as performed here, may provide a means to avoid the rapid loss of fungicide efficacy in the field.

Keywords: blackleg disease, DMI fungicides, fungicide resistance, fungicide sensitivity, in planta assays, population surveys, QOI fungicides, SDHI fungicides.


References

Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M, Parr-Dobrzanski B (2002) The strobilurin fungicides. Pest Management Science 58, 649–662.
The strobilurin fungicides.Crossref | GoogleScholarGoogle Scholar | 12146165PubMed |

Cools HJ, Bayon C, Atkins S, Lucas JA, Fraaije BA (2012) Overexpression of the sterol 14α-demethylase gene (MgCYP51) in Mycosphaerella graminicola isolates confers a novel azole fungicide sensitivity phenotype. Pest Management Science 68, 1034–1040.
Overexpression of the sterol 14α-demethylase gene (MgCYP51) in Mycosphaerella graminicola isolates confers a novel azole fungicide sensitivity phenotype.Crossref | GoogleScholarGoogle Scholar | 22411894PubMed |

Eckert MR, Rossall S, Selley A, Fitt BDL (2010) Effects of fungicides on in vitro spore germination and mycelial growth of the phytopathogens Leptosphaeria maculans and L. biglobosa (phoma stem canker of oilseed rape). Pest Management Science 66, 396–405.
Effects of fungicides on in vitro spore germination and mycelial growth of the phytopathogens Leptosphaeria maculans and L. biglobosa (phoma stem canker of oilseed rape).Crossref | GoogleScholarGoogle Scholar | 20013877PubMed |

Fernández-Ortuño D, Torés JA, de Vicente A, Pérez-García A (2008) Mechanisms of resistance to QoI fungicides in phytopathogenic fungi. International Microbiology 11, 1–9.

Fisher MC, Hawkins NJ, Sanglard D, Gurr SJ (2018) Worldwide emergence of resistance to antifungal drugs challenges human health and food security. Science 360, 739–742.
Worldwide emergence of resistance to antifungal drugs challenges human health and food security.Crossref | GoogleScholarGoogle Scholar | 29773744PubMed |

Fitt BDL, Brun H, Barbetti MJ, Rimmer SR (2006) World-wide importance of phoma stem canker (Leptosphaeria maculans and L. biglobosa) on oilseed rape (Brassica napus. European Journal of Plant Pathology 114, 3–15.
World-wide importance of phoma stem canker (Leptosphaeria maculans and L. biglobosa) on oilseed rape (Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Fraser M, Hwang S-F, Ahmed HU, Akhavan A, Stammler G, Barton W, Strelkov SE (2017) Sensitivity of Leptosphaeria maculans to pyraclostrobin in Alberta, Canada. Canadian Journal of Plant Science 97, 83–91.
Sensitivity of Leptosphaeria maculans to pyraclostrobin in Alberta, Canada.Crossref | GoogleScholarGoogle Scholar |

Garcia-Rubio R, Gonzalez-Jimenez I, Lucio J, Mellado E (2021) Aspergillus fumigatus cross-resistance between clinical and demethylase inhibitor azole drugs. Applied and Environmental Biology 87,
Aspergillus fumigatus cross-resistance between clinical and demethylase inhibitor azole drugs.Crossref | GoogleScholarGoogle Scholar |

Garnault M, Duplaix C, Leroux P, Couleaud G, David O, Walker A-S, Carpentier F (2021) Large-scale study validates that regional fungicide applications are major determinants of resistance evolution in the wheat pathogen Zymoseptoria tritici in France. New Phytologist 229, 3508–3521.
Large-scale study validates that regional fungicide applications are major determinants of resistance evolution in the wheat pathogen Zymoseptoria tritici in France.Crossref | GoogleScholarGoogle Scholar |

Hammond KE, Lewis BG, Musa TM (1985) A systemic pathway in the infection of oilseed rape plants by Leptosphaeria maculans. Plant Pathology 34, 557–565.
A systemic pathway in the infection of oilseed rape plants by Leptosphaeria maculans.Crossref | GoogleScholarGoogle Scholar |

Hollomon DW (2015) Fungicide resistance: facing the challenge: a review. Plant Protection Science 51, 170–176.
Fungicide resistance: facing the challenge: a review.Crossref | GoogleScholarGoogle Scholar |

Ireland KB, Beard C, Cameron J, Chang S, Davidson JA, Dodhia KN, Garrard TA, Hills AL, Hollaway G, Jayasena KW, Kiss L, Mair WJ, Marcroft SJ, McLean MS, Milgate A, Poole N, Simpfendorfer S, Snyman L, Thomas GJ, Wallwork H, Van de Wouw AP, Zulak KG, Lopez-Ruiz FJ (2021) ‘Fungicide resistance management in Australian grain crops’, (Grains Research and Development Corporation: Canberra, ACT, Australia) https://grdc.com.au/fungicide-resistance-management-in-australian-grain-crops. [Accessed August 2021]

Jørgensen LN, Matzen N, Heick TM, Havis N, Holdgate S, Clark B, Glazek M, Blake J, Korbas M, Danielewicz J, Maumene C, Rodemann B, Weigand S, Kildea S, Bataille C, Brauna-Morževska E, Gulbis K, Ban R, Berg G, Semaskiene R, Stammler G (2021) Decreasing azole sensitivity of Z. tritici in Europe contributes to reduced and varying field efficacy. Journal of Plant Diseases and Protection 128, 287–301.
Decreasing azole sensitivity of Z. tritici in Europe contributes to reduced and varying field efficacy.Crossref | GoogleScholarGoogle Scholar |

Joseph-Horne T, Hollomon DW (1997) Molecular mechanisms of azole resistance in fungi. FEMS Microbiology Letters 149, 141–149.
Molecular mechanisms of azole resistance in fungi.Crossref | GoogleScholarGoogle Scholar | 9141655PubMed |

Liu C, Fernando DWG, Gan YT, Kutcher HR, Peng G (2013) Baseline sensitivity of Leptosphaeria maculans to strobilurin fungicides. Canadian Journal of Plant Pathology 35, 119

Ma Z, Proffer TJ, Jacobs JL, Sundin GW (2006) Overexpression of the 14α-demethylase target gene (Cyp51) mediates fungicide resistance in Blumeriella jaapii. Applied and Environmental Microbiology 72, 2581–2585.
Overexpression of the 14α-demethylase target gene (Cyp51) mediates fungicide resistance in Blumeriella jaapii.Crossref | GoogleScholarGoogle Scholar | 16597960PubMed |

Mair WJ, Thomas GJ, Dodhia K, Hills AL, Jayasena KW, Ellwood SR, Oliver RP, Lopez-Ruiz FJ (2020) Parallel evolution of multiple mechanisms for demethylase inhibitor fungicide resistance in the barley pathogen Pyrenophora teres f. sp. maculata. Fungal Genetics and Biology 145, 103475.
Parallel evolution of multiple mechanisms for demethylase inhibitor fungicide resistance in the barley pathogen Pyrenophora teres f. sp. maculata.Crossref | GoogleScholarGoogle Scholar | 33035658PubMed |

Peng G, Liu X, McLaren DL, McGregor L, Yu F (2020) Seed treatment with the fungicide fluopyram limits cotyledon infection by Leptosphaeria maculans and reduces blackleg of canola. Canadian Journal of Plant Pathology 42, 480–492.
Seed treatment with the fungicide fluopyram limits cotyledon infection by Leptosphaeria maculans and reduces blackleg of canola.Crossref | GoogleScholarGoogle Scholar |

Poloni NM, Carvalho G, Vicentini SNC, Dorigan AF, Maciel JLN, McDonald BA, Moreira SI, Hawkins N, Fraaije BA, Kelly DE, Kelly SL, Ceresini PC (2021) Widespread distribution of resistance to triazole fungicides in Brazilian populations of the wheat blast pathogen. Plant Pathology 70, 436–448.
Widespread distribution of resistance to triazole fungicides in Brazilian populations of the wheat blast pathogen.Crossref | GoogleScholarGoogle Scholar |

Price CL, Parker JE, Warrilow AGS, Kelly DE, Kelly SL (2015) Azole fungicides: understanding resistance mechanisms in agricultural fungal pathogens. Pest Management Science 71, 1054–1058.
Azole fungicides: understanding resistance mechanisms in agricultural fungal pathogens.Crossref | GoogleScholarGoogle Scholar | 25914201PubMed |

R Core Team (2019) R: a language and environment for statistical computing. The R Foundation, Vienna, Austria.

Sewell TR, Hawkins NJ, Stotz HU, Huang YJ, Kelly SL, Kelly DE, Fraaije B, Fitt BDL (2017) Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase. Scientific Reports 7, 15849.
Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol 14α-demethylase.Crossref | GoogleScholarGoogle Scholar | 29158527PubMed |

Sierotzki H, Scalliet G (2013) A review of current knowledge of resistance aspects for the next-generation succinate dehydrogenase inhibitor fungicides. Phytopathology 103, 880–887.
A review of current knowledge of resistance aspects for the next-generation succinate dehydrogenase inhibitor fungicides.Crossref | GoogleScholarGoogle Scholar | 23593940PubMed |

Signorell A (2021) ‘DescToola: tools for descriptive statistics. R package version 0.99.41’. (The R Foundation: Vienna, Austria). https://andrisignorell.github.io/DescTools/; https://github.com/AndriSignorell/DescTools/

Sprague SJ, Balesdent M-H, Brun H, Hayden HL, Marcroft SJ, Pinochet X, Rouxel T, Howlett BJ (2006) Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia. European Journal of Plant Pathology 114, 33–40.
Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia.Crossref | GoogleScholarGoogle Scholar |

Sprague SJ, Marcroft SJ, Lindbeck KD, Ware AH, Khangura RK, Van de Wouw AP (2018) Detection, prevalence and severity of upper canopy infection on mature Brassica napus plants caused by Leptosphaeria maculans in Australia. Crop & Pasture Science 69, 65–78.
Detection, prevalence and severity of upper canopy infection on mature Brassica napus plants caused by Leptosphaeria maculans in Australia.Crossref | GoogleScholarGoogle Scholar |

Van de Wouw AP, Marcroft SJ, Howlett BJ (2016) Blackleg disease of canola in Australia. Crop & Pasture Science 67, 273–283.
Blackleg disease of canola in Australia.Crossref | GoogleScholarGoogle Scholar |

Van de Wouw AP, Elliott VL, Chang S, López-Ruiz FJ, Marcroft SJ, Idnurm A (2017) Identification of isolates of the plant pathogen Leptosphaeria maculans with resistance to the triazole fungicide fluquinconazole using a novel in planta assay. PLoS One 12,
Identification of isolates of the plant pathogen Leptosphaeria maculans with resistance to the triazole fungicide fluquinconazole using a novel in planta assay.Crossref | GoogleScholarGoogle Scholar | 29141039PubMed |

Van de Wouw AP, Howlett BJ, Idnurm A (2018) Changes in allele frequencies of avirulence genes in the blackleg fungus, Leptosphaeria maculans, over two decades in Australia. Crop & Pasture Science 69, 20–29.
Changes in allele frequencies of avirulence genes in the blackleg fungus, Leptosphaeria maculans, over two decades in Australia.Crossref | GoogleScholarGoogle Scholar |

Van de Wouw AP, Marcroft SJ, Sprague SJ, Scanlan JL, Vesk PA, Idnurm A (2021) Epidemiology and management of blackleg of canola in response to changing farming practices in Australia. Australasian Plant Pathology 50, 137–149.
Epidemiology and management of blackleg of canola in response to changing farming practices in Australia.Crossref | GoogleScholarGoogle Scholar |

Van de Wouw AP, Marcroft SJ, Howlett BJ (2016) Blackleg disease of canola in Australia. Crop & Pasture Science 67, 273–283.

Wickham H, François R, Henry L, Müller K (2019) dplyr: A grammar of data manipulation. https://CRAN.R-project.org/package=dplyr.

Yang Y, Marcroft SJ, Forsyth LM, Zhao J, Li Z, Van de Wouw AP, Idnurm A (2020) Sterol demethylation inhibitor fungicide resistance in Leptosphaeria maculans is caused by modifications in the regulatory region of ERG11. Plant Disease 104, 1280–1290.
Sterol demethylation inhibitor fungicide resistance in Leptosphaeria maculans is caused by modifications in the regulatory region of ERG11.Crossref | GoogleScholarGoogle Scholar | 32202465PubMed |

Zhang X, Fernando DW (2018) Insights into fighting against blackleg disease of Brassica napus in Canada. Crop & Pasture Science 69, 40–47.
Insights into fighting against blackleg disease of Brassica napus in Canada.Crossref | GoogleScholarGoogle Scholar |

Zheng X, Koopmann B, Ulber B, von Tiedemann A (2020) A global survey on diseases and pests in oilseed rape–current challenges and innovative strategies of control. Frontiers in Agronomy 2, 590908.
A global survey on diseases and pests in oilseed rape–current challenges and innovative strategies of control.Crossref | GoogleScholarGoogle Scholar |