Tropical Plant Pathology

, Volume 44, Issue 2, pp 140–150 | Cite as

Fitness costs associated with G461S mutants of Monilinia fructicola could favor the management of tebuconazole resistance in Brazil

  • Paulo S. F. Lichtemberg
  • Themis J. Michailides
  • Ryan D. Puckett
  • Walmes M. Zeviani
  • Louise L. May De MioEmail author
Original Article


In this study, we investigated the sensitivity components of demethylation fungicide inhibitors (DMI-tebuconazole) for two different sensitivity genotypes of Monilinia fructicola (G461S mutants and wild-type isolates). These components included the fungicide resistance stability and fitness studied over the course of nine and five successive transfers in vitro and ex vivo, respectively, in the absence of fungicide. The results showed no effect of successive transfers on tebuconazole sensitivity of isolates with the G461S mutation for in vitro and ex vivo assays. Athough stable resistance was found, resistant isolates demonstrated a fitness disadvantage relative to the wild-type isolates. The in vitro assay revealed that wild-type isolates had higher germination rates and mycelial growth capacity; and when tested ex vivo, wild-type isolates could produce more spores and larger lesions in peach fruit. The competition studies corroborate the predicted fitness results, where resistant spores were less frequently re-isolated from two pairs of resistant and sensitive mixtures prepared at different ratios. The results obtained in this study have important implications for tebuconazole resistance management of stone-fruit orchards in Brazil and field tests are currently underway to confirm its practicality.


Brown rot disease Demethylation-inhibiting fungicides Peach Sensitivity genotypes 



We thank David Morgan and Michael Luna from the University of California Davis/Kearney Agricultural Research and Extension Center for their technical assistance and Bayer SA for supplying the fungicide used in this study. A special appreciation to the Coordination for the Improvement of Higher Education Personnel (CAPES) doctoral fellowship for making this study possible both in Brazil and in California. This study is based upon work supported by Conselho Nacional de Desenvolvimento Cientifico e Tecnológico grants no 479041/2010-5 Universal/CNPq and grant no 552334/2011-1 CNPQ-Procad.

Supplementary material

40858_2018_254_MOESM1_ESM.pdf (1014 kb)
ESM 1 (PDF 1014 kb)


  1. Adaskaveg J, Schnabel G, Förster H (2008) Diseases of peach caused by fungi and fungal-like organisms: biology, epidemiology and management. In: Layne D, Bassi D (eds) The peach: botany, production and uses. CAB International, Wallingford, pp 352–406CrossRefGoogle Scholar
  2. AGROFIT (2017) Sistema de agrotóxico fitossanitário. Available at: Accessed on January 5th 2018
  3. Anderson JB (2005) Evolution of antifungal-drug resistance: mechanisms and pathogen fitness. Nature Reviews Microbiology 3:547–556CrossRefGoogle Scholar
  4. Bardas G, Myresiotis C, Karaoglanidis G (2008) Stability and fitness of anilinopyrimidine-resistant strains of Botrytis cinerea. Phytopathology 98:443–450CrossRefGoogle Scholar
  5. Box GE, Cox DR (1964) An analysis of transformations. Journal of Royal Statistical Society, Series B 26:211–252Google Scholar
  6. Byrde RJW, Willetts HJ (1977) The brown rot fungi of fruit: their biology and control. Pergamon Press, OxfordGoogle Scholar
  7. Carvalho C, Santos C, Kist B (2017) Anuário brasileiro de fruticultura 2017. Santa Cruz do Sul, Brazil. Editora Gazeta Santa CruzGoogle Scholar
  8. Chen F, Fan J, Zhou T, Liu X, Liu J, Schnabel G (2012) Baseline sensitivity of Monilinia fructicola from China to the DMI fungicide SYP-Z048 and analysis of DMI-resistant mutants. Plant Disease 96:416–422CrossRefGoogle Scholar
  9. Chen F, Liu X, Schnabel G (2013a) Field strains of Monilinia fructicola resistant to both MBC and DMI fungicides isolated from stone fruit orchards in the Eastern United States. Plant Disease 97:1063–1068CrossRefGoogle Scholar
  10. Chen F, Liu X, Chen S, Schnabel E, Schnabel G (2013b) Characterization of Monilinia fructicola strains resistant to both propiconazole and boscalid. Plant Disease 97:645–651CrossRefGoogle Scholar
  11. Chen S, Luo C, Hu M, Schnabel G (2016) Fitness and competitive ability of Botrytis cinerea isolates with resistance to multiple chemical classes of fungicides. Phytopathology 106:997–1005CrossRefGoogle Scholar
  12. Cox K, Bryson P, Schnabel G (2007) Instability of propiconazole resistance and fitness in Monilinia fructicola. Phytopathology 97:448–453CrossRefGoogle Scholar
  13. Elmer P, Braithwaite M, Saville D 1992 Changes in triforine sensitivity in populations of Monilinia fructicola from Hawkes Bay orchards, Proceedings of the New Zealand Plant Protection Conference. New Zealand Plant Protection Society, pp. 138–140Google Scholar
  14. Fischer J, Savi D, Aluizio R, May De Mio L, Glienke C (2017) Characterization of Monilinia species associated with brown rot in stone fruit in Brazil. Plant Pathology 66:423–436CrossRefGoogle Scholar
  15. Gilpatrick J (1981) Resistance to ergosterol biosynthesis-inhibiting fungicides in laboratory strains of Monilinia fructicola. Netherlands Journal Plant Pathology 87:240Google Scholar
  16. Hamilton WD (1964) The genetical evolution of social behaviour. I. Journal of Theoretical Biology 7:1–16CrossRefGoogle Scholar
  17. Hobbelen P, Paveley N, Van den Bosch F (2011a) Delaying selection for fungicide insensitivity by mixing fungicides at a low and high risk of resistance development: a modeling analysis. Phytopathology 101:1224–1233CrossRefGoogle Scholar
  18. Hobbelen P, Paveley N, Fraaije B, Lucas J, Van den Bosch F (2011b) Derivation and testing of a model to predict selection for fungicide resistance. Plant Pathology 60:304–313CrossRefGoogle Scholar
  19. Holmes G, Eckert J (1995) Relative fitness of imazalil-resistant and-sensitive biotypes of Penicillium digitatum. Plant Disease 79:1068–1073CrossRefGoogle Scholar
  20. Hothorn T, Bretz F, Westfall P (2008) Simultaneous inference in general parametric models. Biometrical Journal 50:346–363CrossRefGoogle Scholar
  21. Ishii H (2015) Stability of resistance. In: Ishii H, Hollomon DW (eds) Fungicide resistance in plant pathogens: principles and a guide to practical management. Springer, Japan, pp 35–48CrossRefGoogle Scholar
  22. Karaoglanidis G, Thanassoulopoulos C, Ioannidis P (2001) Fitness of Cercospora beticola field isolates–resistant and–sensitive to demethylation inhibitor fungicides. European Journal of Plant Pathology 107:337–347CrossRefGoogle Scholar
  23. Keske C, Amorim L, May De Mio LL (2011) Peach brown rot incidence related to pathogen infection at different stages of fruit development in an organic peach production system. Crop Protection 30:802–806CrossRefGoogle Scholar
  24. Kim Y, Xiao C (2011) Stability and fitness of pyraclostrobin-and boscalid-resistant phenotypes in field isolates of Botrytis cinerea from apple. Phytopathology 101:1385–1391CrossRefGoogle Scholar
  25. Klosowski AC, Brahm L, Stammler G, De Mio LLM (2016) Competitive fitness of Phakopsora pachyrhizi isolates with mutations in the CYP51 and CYTB genes. Phytopathology 106:1278–1284CrossRefGoogle Scholar
  26. Köller W, Smith F, Reynolds K (1991) Phenotypic instability of flusilazole sensitivity in Venturia inaequalis. Plant Pathology 40:608–611CrossRefGoogle Scholar
  27. Leader J (2004) Numerical analysis and scientific computation. Pearson Addison Wesley, BostonGoogle Scholar
  28. Lichtemberg PSF, Zeviani WM, Michailides TJ, May De Mio LL (2016a) Comparative in vivo and in vitro study on Monilia fructicola causing brown rot of stone fruit in Brazil and California. Tropical Plant Pathology 41:98–106CrossRefGoogle Scholar
  29. Lichtemberg PSF, Zeviani WM, Michailides TJ, May De Mio LL (2016b) Comparison of the sensitivity of Monilinia fructicola isolates to tebuconazole in Brazil using three methods. Canadian Journal of Plant Pathology 38:55–63CrossRefGoogle Scholar
  30. Lichtemberg P, Yong L, Morales R, Muehlmann-Fischer J, Michailides T, May de Mio L (2017) The point mutation G461S in the MfCYP51 gene is associated with tebuconazole resistance in Monilinia fructicola populations in Brazil. Phytopathol 107:1507–1514CrossRefGoogle Scholar
  31. Lim T-H, Chang T-H, Byeongjin C (1999) Biological characteristics of benzimidazole-resistant and-senstive isolates of Monilinia fructicola from peach fruits in Korea. The Plant Pathology Journal 15:340–344Google Scholar
  32. Lim T, Yi J, Chang T, Byeongjin C (2001) Fitness of dicarboximide-resistant and sensitive Monilinia fructicola isolated from peach in Korea. The Plant Pathology Journal 17:205–209Google Scholar
  33. Luo CX, Cox KD, Amiri A, Schnabel G (2008) Occurrence and detection of the DMI resistance-associated genetic element 'Mona' in Monilinia fructicola. Plant Disease 92:1099–1103CrossRefGoogle Scholar
  34. May De Mio LL, Garrido L, Ueno B (2004) Doenças de fruteiras de caroço. In: Monteiro LB, May De Mio LL, Serrat BM, Motta AC, Cuquel FL (eds) Fruteiras de caroço: Uma visão ecológica. Curitiba, UFPR, pp 169–222Google Scholar
  35. May De Mio LL, Luo Y, Michailides TJ (2011) Sensitivity of Monilinia fructicola from Brazil to tebuconazole, azoxystrobin, and thiophanate-methyl and implications for disease management. Plant Disease 95:821–827CrossRefGoogle Scholar
  36. Moreira LM, May De Mio LL, Valdebenito-Sanhueza RM (2008) Fungos antagonistas e efeito de produtos químicos no controle da podridão parda em pomar de pessegueiro. Summa Phytopathologica 34:272–276CrossRefGoogle Scholar
  37. Nikou D, Malandrakis A, Konstantakaki M, Vontas J, Markoglou A, Ziogas B (2009) Molecular characterization and detection of overexpressed C-14 alpha-demethylase-based DMI resistance in Cercospora beticola field isolates. Pesticide Biochemistry and Physiology 95:18–27CrossRefGoogle Scholar
  38. Nuninger-Ney C, Schwinn FJ, Staub T (1989) In vitro selection of sterol-biosynthesis-inhibitor (SBI)-resistant mutants in Monilinia fructicola (Wint.) honey. Netherlands Journal of Plant Pathology 95:137–150Google Scholar
  39. Oliver RP, Hewitt HG (2014) Fungicides in crop protection. Second ed. Oxfordshire, UK. CAB InternationalGoogle Scholar
  40. Peever TL, Milgroom MG (1994) Lack of correlation between fitness and resistance to sterol biosynthesis-inhibiting fungicides in Pyrenophora teres. Phytopathology 84:515–519CrossRefGoogle Scholar
  41. Penrose L, Davis K, Koffmann W (1979) The distribution of benomyl-tolerant Sclerotinia fructicola (Wint.) Rehm. In stone fruit orchards in New South Wales and comparative studies with susceptible isolates. Crop Pasture &Science 30:307–319CrossRefGoogle Scholar
  42. Pfeufer EE, Ngugi HK (2012) Orchard factors associated with resistance and cross resistance to sterol demethylation inhibitor fungicides in populations of Venturia inaequalis from Pennsylvania. Phytopathology 102:272–282CrossRefGoogle Scholar
  43. Primiano IV, Molina JPE, Mio LLMD, Peres NA, Amorim L (2017) Reduced sensitivity to azoxystrobin is stable in Monilinia fructicola isolates. Scientia Agricola 74:169–173CrossRefGoogle Scholar
  44. Sanoamuang N, Gaunt R (1995) Persistence and fitness of carbendazim-and dicarboximide-resistant isolates of Monilinia fructicola (Wint.) honey in flowers, shoots and fruit of stone fruit. Plant Pathology 44:448–457CrossRefGoogle Scholar
  45. Schepers H (1985) Fitness of isolates of Sphaerotheca fuliginea resistant or sensitive to fungicides which inhibit ergosterol biosynthesis. Netherlands Journal of Plant Pathology 91:65–76CrossRefGoogle Scholar
  46. Schnabel G, Jones AL (2001) The 14α-demethylasse (CYP51A1) gene is overexpressed in Venturia inaequalis strains resistant to myclobutanil. Phytopathology 91:102–110CrossRefGoogle Scholar
  47. Silva S, Kohls V, Manica-Berto R, Rigatto P, Rombaldi C (2011) Apropriação tecnológica da produção integrada de pêssego na região de Pelotas no estado do Rio Grande do Sul. Ciência Rural 41:1667–1673CrossRefGoogle Scholar
  48. Smith FD, Parker DM, Köller W (1991) Sensitivity distribution of Venturia inaequalis to the sterol demethylation inhibitor flusilazole: baseline sensitivity and implications for resistance monitoring. Phytopathology 81:392–396CrossRefGoogle Scholar
  49. van Den Bosch F, Paveley N, Shaw M, Hobbelen P, Oliver R (2011) The dose rate debate: does the risk of fungicide resistance increase or decrease with dose? Plant Pathology 60:597–606CrossRefGoogle Scholar
  50. van den Bosch F, Paveley N, van den Berg F, Hobbelen P, Oliver R (2014) Mixtures as a fungicide resistance management tactic. Phytopathology 104:1264–1273CrossRefGoogle Scholar
  51. Vega B, Dewdney MM (2014) QoI-resistance stability in relation to pathogenic and saprophytic fitness components of Alternaria alternata from citrus. Plant Disease 98:1371–1378CrossRefGoogle Scholar
  52. Villani SM, Cox KD (2011) Characterizing Fenbuconazole and Propiconazole sensitivity and prevalence of 'Mona'in isolates of Monilinia fructicola from New York. Plant Disease 95:828–834CrossRefGoogle Scholar
  53. Zanette F, Biasi L (2004) Introdução a fruteiras de caroço. In: Monteiro LB, May De Mio LL, Serrat BM, Motta AC, Cuquel FL (eds) Fruteiras de caroço: Uma visão ecológica. Curitiba, UFPR, pp 1–4Google Scholar
  54. Zhu F, Bryson PK, Schnabel G (2012) Influence of storage approaches on instability of propiconazole resistance in Monilinia fructicola. Pest Management Science 68:1003–1009CrossRefGoogle Scholar

Copyright information

© Sociedade Brasileira de Fitopatologia 2018

Authors and Affiliations

  1. 1.Department of Plant PathologyFederal University of Paraná StateCuritibaBrazil
  2. 2.Department of Plant PathologyUniversity of California-DavisParlierUSA
  3. 3.Department of StatisticsFederal University of Paraná StateCuritibaBrazil

Personalised recommendations