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Sensitivity of Cercospora spp. from soybean to quinone outside inhibitors and methyl benzimidazole carbamate fungicides in Brazil

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Abstract

Cercospora leaf blight and purple seed stain are caused by Cercospora kikuchii and other Cercospora spp. The fungus can infect leaves and seeds on soybean, and the disease is controlled with fungicides. In Brazil, the intensive use of fungicides on soybean to control other diseases such as soybean rust has exerted resistance selection pressure on all fungal pathogens that attack in the crop. In this study, we evaluated the sensitivity of 56 Cercospora spp. soybean isolates collected during  9 crop seasons in 9 Brazilian states to quinone outside inhibitors (QoI) and methyl benzimidazole carbamate (MBC) fungicides using mycelial growth inhibition in amended media with a discriminatory dose of 10 μg/mL. We also analyzed single polymorphisms in the target genes cytb and β-tubulin using target genotyping by sequencing by Illumina short reads. Genome-local association was used to correlate the point mutations found in the coding sequence with in vitro assays results. For cytb gene, it was observed only the G143A mutation in 73% of the isolates. The mutation G143A was present in 97% of the isolates classified as resistant at least for two QoI fungicides. The mutation E198A was present in 71% of the isolates being significantly associated with the reduction control to MBC fungicides. Cross-resistance was observed into QoI (azoxystrobin, picoxystrobin, and pyraclostrobin) and MBC (carbendazim, and thiophanate-methyl) active ingredients fungicides. About 97% of Cercospora spp. isolates analyzed possessed the double mutations G143A and E198A and resulting in less sensitive to QoI and MBC fungicides, respectively. All isolates showed homozygous variation in the alternative allele in G143A and E198A mutations. Our results suggest that resistance of the Cercospora spp. from soybean to QoI and MBC fungicides has occurred since the 2008 cropping season in Brazil associated with G143A and E198A mutations.

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Data availability

Data that support the findings of this study have been deposited in the National Center for Biotechnology Information (NCBI) under accession number PRJNA634433.

References

  • AGROFIT. Consulta de ingredientes ativos 2019. Available at: : http:// http://agrofit.agricultura.gov.br/agrofit_cons/principal_agrofit_cons. Accessed on: 09 Jan 2020

  • Albu S, Price T, Doyle V, Padgett B, Schneider R (2016) The G143A mutation is responsible for strobilurin fungicide resistance in Cercospora cf. flagellaris, a leaf blight and purple seed stain pathogen of Louisiana soybean. Plant Health Progress 17:197–197

    Article  Google Scholar 

  • Andrews S (2010) FastQC: a quality control tool for high throughput sequence data. Available at: http://www.bioinformatics.babraham.ac.uk/projects/fastqc/. Accessed on: 05 Aug 2020

  • Birla K, Rivera-Varas V, Secor GA, Khan MF, Bolton MD (2012) Characterization of cytochrome b from European field isolates of Cercospora beticola with quinone outside inhibitor resistance. European Journal of Plant Pathology 134:475–488

    Article  CAS  Google Scholar 

  • Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120

    Article  CAS  Google Scholar 

  • Bolton MD, Rivera V, Secor G (2013) Identification of the G143A mutation associated with QoI resistance in Cercospora beticola field isolates from Michigan, United States. Pest Management Science 69:35–39

    Article  CAS  Google Scholar 

  • Bradley CA, Pedersen DK (2011) Baseline sensitivity of Cercospora zeae-maydis to quinone outside inhibitor fungicides. Plant Disease 95:189–194

    Article  CAS  Google Scholar 

  • CONAB. Acompanhamento da safra brasileira de grãos, v. 7 - Safra 2019/20, n.4 - Sexto levantamento, janeiro 2020. Available at: https://www.conab.gov.br/info-agro/safras/graos. Accessed on 09 Jan 2020

  • CONSÓRCIO ANTIFERRUGEM (2019) Custo ferrugem asiática da soja . Available at: http://www.consorcioantiferrugem.net/#/conteudos/view/5. Accessed on 8 Feb 2020

  • Danecek P, Auton A, Abecasis G, Albers CA, Banks E, DePristo MA, Handsaker RE, Lunter G, Marth GT, Sherry ST, McVean G (2011) The variant call format and VCFtools. Bioinformatics 27:2156–2158

    Article  CAS  Google Scholar 

  • Duan Y, Xin W, Lu F, Li T, Li M, Wu J, Wang J, Zhou M (2019) Benzimidazole-and QoI-resistance in Corynespora cassiicola populations from greenhouse-cultivated cucumber: an emerging problem in China. Pesticide Biochemistry and Physiology 153:95–105

    Article  CAS  Google Scholar 

  • FRAC. List of plant pathogenic organisms resistant to disease control agents. Available at: https://www.frac.info/docs/default-source/publications/list-of-resistant-plant-pathogens/list-of-resistant-plant-pathogenicorganisms_may2018.pdf?sfvrsn=a2454b9a_2. Accessed on: 08 Jan 2020

  • Gisi U, Sierotzki H, Cook A, McCaffery A (2002) Mechanisms influencing the evolution of resistance to Qo inhibitor fungicides. Pest Management Science 58:859–867

    Article  CAS  Google Scholar 

  • Godoy CV, Almeida AMR, Costamilan LM, Meyer MC, Dias WP, Seixas CDS, Soares RM, Henning AA, Yorinori JT, Ferreira LP, Silva JFV (2016) Doenças da soja. In: Amorim L, Rezende JAM, Bergamin Filho A, Camargo LEA (eds) Manual de Fitopatologia. Vol. 2, Doenças das plantas cultivadas, vol 5. Ed. Ceres, São Paulo, pp 657–675

    Google Scholar 

  • Hartman GL, Rupe JC, Sikora EJ, Domier LL, Davis JA, Steffey KL (eds) (2015) Compendium of soybean diseases and pests. APS Press, The American Phytopathological Society, St. Paul

    Google Scholar 

  • Imazaki I, Iizumi H, Ishikawa K, Sasahara M, Yasuda N, Koizumi S (2006) Effects of thiophanate-methyl and azoxystrobin on the composition of Cercospora kikuchii populations with thiophanate-methyl-resistant strains. Journal of General Plant Pathology 72:292–300

    Article  CAS  Google Scholar 

  • Ishii H, Fountaine J, Chung WH, Kansako M, Nishimura K, Takahashi K, Oshima M (2009) Characterisation of QoI-resistant field isolates of Botrytis cinerea from citrus and strawberry. Pest Management Science: formerly Pesticide Science 65:916–922

    Article  CAS  Google Scholar 

  • Kayamori M, Shimizu M, Yamana T, Komatsu T, Minako S, Shinmura A, Sasaki J, Kozawa T, Notsu A, Yasuoka S (2020) First report of QoI resistance in Cercospora beticola in sugar beet in Japan. Journal of General Plant Pathology 86:149–153

    Article  CAS  Google Scholar 

  • Klosowski AC, May De Mio LL, Miessner S, Rodrigues R, Stammler G (2016) Detection of the F129L mutation in the cytochrome b gene in Phakopsora pachyrhizi. Pest Management Science 72:1211–1215

    Article  CAS  Google Scholar 

  • Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nature Methods 9:357–359

    Article  CAS  Google Scholar 

  • Lipka AE, Tian F, Wang Q, Peiffer J, Li M, Bradbury PJ, Zhang Z (2012) GAPIT: genome association and prediction integrated tool. Bioinformatics 28:2397–2399

    Article  CAS  Google Scholar 

  • Ma Z, Michailides TJ (2005) Advances in understanding molecular mechanisms of fungicide resistance and molecular detection of resistant genotypes in phytopathogenic fungi. Crop Protection 24:853–863

    Article  CAS  Google Scholar 

  • Mathew FM, Byamukama E, Neves DL, Bradley CA (2019) Resistance to quinone outside inhibitor fungicides conferred by the G143A mutation in Cercospora sojina (causal agent of frogeye leaf spot) isolates from South Dakota soybean fields. Plant Health Progress 20:104–105

    Article  Google Scholar 

  • McGrath MT (2015) Cucurbit powdery mildew in the USA. In: Fungicide resistance in plant pathogens. Springer,Tokyo. p. 401–417

  • Price PP III, Purvis MA, Cai G, Padgett GB, Robertson CL, Schneider RW, Albu S (2015) Fungicide resistance in Cercospora kikuchii, a soybean pathogen. Plant Disease 99:1596–1603

    Article  CAS  Google Scholar 

  • R Core Team (2019) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available at: URL https://www.R-project.org/. Accessed on: 05 Feb 2020

  • Recomendações técnicas para a cultura da soja na cultura da soja na região central do Brasil 2000/01 (2000) Embrapa Soja/ Fundação MT, Londrina PR. p. 245

  • Rimmer A, Phan H, Mathieson I, Iqbal Z, Twigg SR, Wilkie AO, McVean G, Lunter G (2014) Integrating mapping-, assembly-and haplotype-based approaches for calling variants in clinical sequencing applications. Nature Genetics 46:912–918

    Article  CAS  Google Scholar 

  • Sautua, FJ, Gonzalez SA, Doyle VP, Berretta MF, Gordó M, Scandiani MM, Rivarola ML, Fernandez P, Carmona MA (2019a) Draft genome sequence data of Cercospora kikuchii, a causal agent of Cercospora leaf blight and purple seed stain of soybeans. Data in Brief 27:104693

  • Sautua FJ, Searight J, Doyle VP, Price PP III, Scandiani MM, Carmona MA (2019b) The G143A mutation confers azoxystrobin resistance to soybean Cercospora leaf blight in Bolivia. Plant Health Progress 20:2–3

    Article  Google Scholar 

  • Sautua FJ, Doyle VP, Price PP, Porfiri A, Fernandez P, Scandiani MM, Carmona MA (2020) Fungicide resistance in Cercospora species causing cercospora leaf blight and purple seed stain of soybean in Argentina. Plant Pathology 00:1–17

    Google Scholar 

  • Soares AP, Guillin EA, Borges LL, Da Silva AC, De Almeida ÁM, Grijalba PE, Gottlieb AM, Bluhm BH, De Oliveira LO (2015) More Cercospora species infect soybeans across the Americas than meets the eye. PLoS One 10:e0133495

    Article  Google Scholar 

  • Wickham H (2016) ggplot2: elegant graphics for data analysis. Available at: https://ggplot2.tidyverse.org. Accessed on 30 July 2020

  • Zeng F, Arnao E, Zhang G, Olaya G, Wullschleger J, Sierotzki H, Ming R, Bluhm BH, Bond JP, Fakhoury AM, Bradley CA (2015) Characterization of quinone outside inhibitor fungicide resistance in Cercospora sojina and development of diagnostic tools for its identification. Plant Disease 99:544–550

    Article  CAS  Google Scholar 

  • Zhou T, Mehl HL (2020) Rapid quantification of the G143A mutation conferring fungicide resistance in Virginia populations of Cercospora sojina using pyrosequencing. Crop Protection 127:104942

    Article  CAS  Google Scholar 

Download references

Funding

The first author thanks the Jeff Schell scholarship by Bayer Foundations and Coordination of Superior Level Staff Improvement (CAPES) scholarship for funding this research.

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Authors and Affiliations

  1. Universidade Estadual de Londrina, Londrina, Paraná, 86051-970, Brazil

    Flávia E. de Mello, Helen Prudente, Sheila A. Xavier-Valencio & Maria I. Balbi-Peña

  2. Embrapa Soja, Londrina, Paraná, 86001-970, Brazil

    Valéria S. Lopes-Caitar, Francismar C. Marcelino-Guimaraes & Cláudia V. Godoy

  3. University of Kiel, 24118, Kiel, Germany

    Sören Franzenburg & Joseph-Alexander Verreet

  4. Bayer CropScience, 40789, Monheim, Germany

    Andreas Mehl

Authors
  1. Flávia E. de Mello
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  2. Valéria S. Lopes-Caitar
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  3. Helen Prudente
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  4. Sheila A. Xavier-Valencio
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  5. Sören Franzenburg
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  6. Andreas Mehl
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  7. Francismar C. Marcelino-Guimaraes
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  8. Joseph-Alexander Verreet
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  9. Maria I. Balbi-Peña
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  10. Cláudia V. Godoy
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Contributions

CVG, FCMG, MIBP, AM, and JAV planned and designed the experimental work. FEM, HP, SAXV, and SF executed the experiments. FEM and VSLC conducted the data analyses. FEM, VSLC, and CVG wrote the manuscript.

Corresponding author

Correspondence to Flávia E. de Mello.

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Authors declare that this manuscript have not published elsewhere. All authors read and approved the final version of this manuscript. The authors declare that the present work was developed without any potential conflict of interest, with no human or animal participants.

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de Mello, F.E., Lopes-Caitar, V.S., Prudente, H. et al. Sensitivity of Cercospora spp. from soybean to quinone outside inhibitors and methyl benzimidazole carbamate fungicides in Brazil. Trop. plant pathol. 46, 69–80 (2021). https://doi.org/10.1007/s40858-020-00410-4

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