Skip to main content
Log in

Development of benzimidazole resistant Trichoderma strains for the integration of chemical and biocontrol methods of grapevine pruning wound protection

  • Published:
BioControl Aims and scope Submit manuscript

Abstract

In the protection of grapevine pruning wounds from trunk pathogen infection, fungicides provide mainly short term protection while biocontrol agents provide mainly long term protection. The integration of fungicide and biological wound protection could provide better wound protection, but is limited by the susceptibility of the biocontrol agents to the fungicides. Stable benzimidazole resistant mutants were generated by gamma irradiation (250 Gy) from three wild-type Trichoderma Pers. isolates (UST1, UST2 and T77) shown to provide wound protection. The wild-type Trichoderma isolates were found to be naturally resistant to thiophanate methyl while mycelial growth was completely inhibited by 2.5 μg ml−1 of benomyl and carbendazim. There was no reduction in biological fitness and in vitro antagonist activity for mutants generated from UST1 and UST2 while the mutant from T77 had reduced fitness and antagonistic activity compared to its wild type. The wild type and the mutant of UST1 were further tested in the field and significantly (P < 0.001) reduced pruning wound infection by Phaeomoniella chlamydospora (W. Gams, Crous, M.J. Wingf. & Mugnai) Crous & W. Gams, when applied alone and in combination with thiophanate methyl and carbendazim, respectively. The combination of the mutant UST1 with carbendazim gave the highest reduction of infection compared to the rest of the treatments when P. chlamydospora was inoculated 24 h after pruning. The Trichoderma transformants generated in this study can be applied in combination with benzimidazole fungicides for a more effective and sustainable wound protection.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1

Similar content being viewed by others

References

  • Ahloowalia BS, Maluszynski M (2001) Induced mutations—a new paradigm in plant breeding. Euphytica 11:167–173

    Article  Google Scholar 

  • Blank G, Corrigan D (1995) Comparison of resistance of fungal spores to gamma and electron beam radiation. Int J Food Microbiol 26:269–277

    Article  CAS  PubMed  Google Scholar 

  • Braghini R, Pozzi CR, Aquino S, Rocha LO, Corrêa B (2009) Effects of gamma-radiation on the fungus Alternaria alternata in artificially inoculated cereal samples. Appl Radiat Isot 67:1622–1628

    Article  CAS  PubMed  Google Scholar 

  • Brunner K, Zeilinger S, Ciliento R, Woo SL, Lorito M, Kubicek CP, Mach RL (2005) Improvement of the fungal biocontrol agent Trichoderma atroviride to enhance both antagonism and induction of plant systemic disease resistance. Appl Environ Microbiol 71:3959–3965

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Carter M, Price T (1975) Biological control of Eutypa armeniacae. III A comparison of chemical, biological and integrated control. Aust J Agric Res 26:537–543

    Article  CAS  Google Scholar 

  • Chapuis L, Richard L, Dubos B (1998) Variation in susceptibility of grapevine pruning wound to infection by Eutypa lata in south-western France. Plant Pathol 47:463–472

    Article  Google Scholar 

  • Creaser ML, Wicks T (2002) Evaluation of fungicides, paints and a biological product for the prevention of Eutypa dieback of grapevines. Aust NZ Grapegrow Winemak 465:15–18

    Google Scholar 

  • Crous PW, Gams W (2000) Phaeomoniella chlamydospora gen. Et comb. Nov., a causal organism of Petri grapevine decline and esca. Phytopathol Mediterr 39:112–188

    Google Scholar 

  • Dadachova E, Casadevall A (2008) Ionizing radiation: how fungi cope, adapt, and exploit with the help of melanin. Curr Opin Microbiol 11:525–531

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Díaz GA, Latorre BA (2013) Efficacy of paste and liquid fungicide formulations to protect pruning wounds against pathogens associated with grapevine trunk diseases in Chile. Crop Prot 46:106–112

    Article  Google Scholar 

  • Faretra F, Pollastro S (1991) Genetic basis of resistance to benzimidazole and dicarboximide fungicides in Botryotinia fuckeliana (Botrytis cinerea). Mycol Res 95:943–951

    Article  CAS  Google Scholar 

  • Fourie PH, Halleen F (2004) Proactive control of Petri disease of grapevine through treatment of propagation material. Plant Dis 88:1241–1245

    Article  Google Scholar 

  • Fourie PH, Halleen F (2006) Chemical and biological protection of grapevine propagation material from trunk disease pathogens. Eur J Plant Pathol 116:255–265

    Article  CAS  Google Scholar 

  • Fourie PH, Halleen F, van der Vyver J, Schrueder W (2001) Effect of Trichoderma treatments on the occurrence of decline pathogens on the roots and rootstocks of nursery plants. Phytopathol Mediterr 40S:473–478

    Google Scholar 

  • Gendloff EH, Ramsdell DC, Burton C (1983) Fungicidal control of Eutypa armeniacae infecting concord grapevine in Michigan. Plant Dis 67:754–756

    Article  CAS  Google Scholar 

  • Gramaje D, Armengol J (2011) Fungal trunk pathogens in the grapevine propagation process: potential inoculum sources, detection, identification and management strategies. Plant Dis 95:1040–1055

    Article  Google Scholar 

  • Gramaje D, Aroca A, Raposo R, García-Jiménez J, Armengol J (2009) Evaluation of fungicides to control Petri disease pathogens in the grapevine propagation process. Crop Prot 28:1091–1097

    Article  CAS  Google Scholar 

  • Gramaje D, Ayres MR, Trouillas FP, Sosnowski MR (2012) Efficacy of fungicides on mycelial growth of diatrypaceous fungi associated with grapevine trunk disease. Australas Plant Pathol 41:295–300

    Article  CAS  Google Scholar 

  • Gubler WD, Rolshausen PE, Trouillas FP, Úrbez-Torres JR, Voegel TM, Leavitt GM, Weber EA (2005) Grapevine trunk diseases in California. Pract Winery Vineyard. Jan/Feb, 6–25

  • Halleen F, Crous PW, Petrini O (2003) Fungi associated with healthy grapevine cuttings in nurseries, with special reference to pathogens involved in the decline of young vines. Australas Plant Pathol 32:47–52

    Article  Google Scholar 

  • Halleen F, Fourie PH, Lombard PJ (2010) Protection of grapevine pruning wounds against Eutypa lata by biological and chemical methods. S Afr J Enol Vitic 31:125–132

    CAS  Google Scholar 

  • Hanson L, Howell C (2002) Biocontrol efficacy and other characteristics of protoplast fusants between Trichoderma koningii and T. virens. Mycol Res 106:321–328

    Article  Google Scholar 

  • Harman G (2000) Myths and dogmas of biocontrol changes in perceptions derived from research on Trichoderma harzinum T-22. Plant Dis 84:377–393

    Article  Google Scholar 

  • Hatvani L, Manczinger L, Kredics L, Szekeres A, Antal Z, Vágvölgyi C (2006) Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion. Antonie van Leeuwenhoek 89:387–393

    Article  PubMed  Google Scholar 

  • John S, Wicks TJ, Hunt JS, Lorimer MF, Oakey H, Scott ES (2005) Protection of grapevine pruning wounds from infection by Eutypa lata using Trichoderma harzianum and Fusarium lateritium. Australas Plant Pathol 34:569–575

    Article  Google Scholar 

  • Keinath A, Zitter T (1998) Resistance to benomyl and thiophanate-methyl in Didymella bryoniae from South Carolina and New York. Plant Dis 82:479–484

    Article  CAS  Google Scholar 

  • Kotze C, van Niekerk J, Halleen F, Fourie PH (2011) Evaluation of biocontrol agents for grapevine pruning wound protection against trunk pathogen infection. Phytopathol Mediterr 50(Supplement):S247–S263

    Google Scholar 

  • Kovács E, Keresztes A (2002) Effect of gamma and UV-B/C radiation on plant cells. Micron 33:199–210

    Article  PubMed  Google Scholar 

  • Larignon P, Dubos B (2000) Preliminary studies on the biology of Phaeoacremonium. Phytopathol Mediterr 39:184–189

    Google Scholar 

  • Leroux P, Chapeland F, Desbrosses D, Gredt M (2000) Patterns of cross-resistance to fungicides in Botryotinia fuckeliana (Botrytis cinerea) isolates from French vineyards. Crop Prot 18:687–697

    Article  Google Scholar 

  • Lewis JA, Papavizas GC (1991) Biocontrol of plant diseases: the approach for tomorrow. Crop Prot 10:95–105

    Article  Google Scholar 

  • Ma Z, Yoshimura M, Michailides T (2003) Identification and characterization of benzimidazole resistance in Monilinia fructicola from stone fruit orchards in California. Appl Environ Microbiol 69:7145–7152

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Matthews RW (1982) Effect of temperature on the radiation induced reduction of dichromate ion. Int J Radiat Isot 33:454–455

    Article  CAS  Google Scholar 

  • Moeller R, Stackebrandt E, Reitz G, Berger T, Rettberg P, Doherty AJ, Horneck G, Nicholson WL (2007) Role of DNA repair by nonhomologous-end joining in Bacillus subtilis spore resistance to extreme dryness, mono- and polychromatic UV, and ionizing radiation. J Bacteriol 189:3306–3311

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Moller W, Kasimatis A (1980) Protection of grapevine pruning wounds from Eutypa dieback. Phytopathology 64:278–280

    Google Scholar 

  • Mostert L, Groenewald JZ, Summerbell RC, Gams W, Crous PW (2006) Taxonomy and pathology of Togninia (Diaporthales) and its Phaeoacremonium anamorphs. Stud Mycol 54:1–113

    Article  Google Scholar 

  • Mugnai L, Graniti A, Surico G (1999) Esca (black measles) and brown wood-streaking: two old and elusive diseases of grapevines. Plant Dis 83:404–418

    Article  Google Scholar 

  • Munkvold G, Marois J (1993a) The effects of fungicides on Eutypa lata germination, growth and infection of grapevines. Plant Dis 77:50–55

    Article  CAS  Google Scholar 

  • Munkvold G, Marois J (1993b) Efficacy of natural epiphytes and colonizers of grapevine pruning wounds for biological control of Eutypa dieback. Phytopathology 83:624–629

    Article  Google Scholar 

  • Mutawila C, Fourie PH, Halleen F, Mostert L (2011) Histo-pathology study of the growth of Trichoderma harzianum, Phaeomoniella chlamydospora and Eutypa lata on grapevine pruning wounds. Phytopathol Mediterr 50(Supplement):S46–S60

    Google Scholar 

  • Ossanna N, Mischke S (1990) Genetic transformation of the biocontrol fungus Gliocladium virens to benomyl resistance. Appl Environ Microbiol 56:3052–3056

    PubMed Central  CAS  PubMed  Google Scholar 

  • Paparu P, Mcleod A, Dubois T, Coyne D, Viljoen A (2009) Efficacy of chemical and fluorescent protein markers in studying plant colonization by endophytic non-pathogenic Fusarium oxysporum isolates. BioControl 54:709–722

    Article  CAS  Google Scholar 

  • Pascoe I, Cottral E (2000) Developments in grapevine trunk diseases research in Australia. Phytopathol Mediterr 39:68–75

    Google Scholar 

  • Petzoldt C, Moller W, Sall M (1981) Eutypa dieback of grapevine: seasonal differences in infection and duration of susceptibility of pruning wounds. Phytopathology 71:540–543

    Article  Google Scholar 

  • Rolshausen P, Gubler WD (2005) Use of boron for the control of Eutypa dieback of gapevines. Plant Dis 89:734–738

    Article  CAS  Google Scholar 

  • Rolshausen PE, Úrbez-Torres JR, Rooney-Latham S, Eskalen A, Smith RJ, Gubler WD (2010) Evaluation of pruning wound susceptibility and protection against fungi associated with grapevine trunk diseases. Am J Enol Vitic 61:113–119

    Google Scholar 

  • Serra S, Mannoni MA, Ligios V (2008) Studies on the susceptibility of pruning wounds to infection by fungi involved in grapevine wood diseases in Italy. Phytopathol Mediterr 47:234–246

    Google Scholar 

  • Surico G, Mugnai L, Marchi G (2008) The ecsa complex. In: Ciancio A, Mukerji K (eds) Integrated management of diseases casued by fungi, phytoplasma and bacteria. Springer, Dordrecht, The Netherlands, pp 119–136

  • Trouillas FP, Úrbez-Torres JR, Gubler WD (2010) Diversity of diatrypaceous fungi associated with grapevine canker diseases in California. Mycologia 102:319–336

    Article  CAS  PubMed  Google Scholar 

  • van Niekerk JM, Crous PW, Groenwald JZ, Fourie PH, Halleen F (2004) DNA phylogeny, morphology and pathogenicity of Botryosphaeria species on grapevines. Mycologia 96:781–798

    Article  PubMed  Google Scholar 

  • van Niekerk JM, Groenwald JZ, Farr C, Fourie PH, Halleen F, Crous PW (2005) Reassessment of Phomopsis species on grapevine. Austaralas Plant Pathol 34:27–39

    Article  Google Scholar 

  • van Niekerk J, Halleen F, Fourie P (2011) Temporal susceptibility of grapevine pruning wounds to trunk pathogen infection in South African grapevines. Phytopathol Mediterr 50 (Supplement):S139–S150

    Google Scholar 

  • World Health Organisation (WHO) (1994) Safety and nutritional adequacy of irradiated food. World Health Organisation, Geneva, p 161. Accessible on: http://www.who.int/iris/handle/10665/39463. Accessed 12 Mar 2013

  • Yan K, Dickman M (1996) Isolation of a beta-tubulin gene from Fusarium moniliforme that confers cold-sensitive benomyl resistance. Appl Environ Microbiol 62:3053–3056

    PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We acknowledge financial support from the South African Table Grape Industry (SATI), Winetech, the Technology and Human Resources for Industry Programme (THRIP) and National Research Foundation (NRF). The authors appreciate the technical assistance that was provided by Dr. J.P. Slabbert and Mr. P. Beukes of iThemba Labs, Cape Town for the irradiation dosimetry and fungal irradiation.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lizel Mostert.

Additional information

Handling Editor: Jesus Mercado Blanco.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mutawila, C., Halleen, F. & Mostert, L. Development of benzimidazole resistant Trichoderma strains for the integration of chemical and biocontrol methods of grapevine pruning wound protection. BioControl 60, 387–399 (2015). https://doi.org/10.1007/s10526-014-9647-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10526-014-9647-y

Keywords

Navigation