Abstract
Trichoderma aggressivum is an aggressive contaminant mould in the cultivation of Agaricus bisporus leading to severe reductions in mushroom yields. Production of fully colonised A. bisporus substrate in Europe is commonly carried out in large tunnels (Phase III), after which the substrate undergoes several bulk handling (mixing) operations before ending up on shelves in mushroom growing facilities. The work presented here studied the effect of Trichoderma aggressivum inoculum, substrate mixing and supplementation on Agaricus bisporus yields and evaluated four methods to detect T. aggressivum in bulk handled substrate. Inoculum dilution level was shown to correlate well with mushroom yield (P < 0.0001) with reductions of 2–6 % at the most dilute level (10−4) and 60–100 % at the most concentrated level (10−1), depending on the experiment. Supplementation, with or without T. aggressivum, had no significant effect on mushroom yield (P ≥ 0.85) but a high degree of substrate mixing was shown to significantly increase (P < 0.0001) T. aggressivum-associated crop losses. Four T. aggressivum detection methods were evaluated and a quantitative polymerase chain reaction (qPCR) method gave the most consistent and least variable results. Cycle threshold (CT) values ranged from 24 to 40, depending on the experiment and the inoculum dilution level, and false negatives (CT = 40) were reported on one occasion with the most dilute samples. The results indicate that Phase III mushroom substrate is vulnerable to infection by T. aggressivum when the fully colonised substrate is broken up and mixed during bulk handling operations, identifying a previously unidentified risk for Phase III substrate producers.
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References
Anon. (2016a). MushComb Flexible New Machinery. http://issuu.com/willigendael/docs/mush_comb_brochure
Anon. (2016b). Hoving Holland Int. BV. Mushroom and composting equipment. http://www.hovingholland.com/muschroom-culture-machinery-manufacturer-treatment-cultivation
Beyer, D. M., Wuest, P. J., & Kremser, J. J. (2000). Evaluation of epidemiological factors and mushroom substrate characteristics influencing the occurrence and development of Trichoderma green mold. Mushroom Science, 15(2), 633–640.
Briones Jr., M. A., & Reichardt, W. (1999). Estimating microbial population counts by most probable number using Microsoft excel®. Journal of Microbiological Methods, 35, 157–161.
Chen, X., Romaine, C. P., Ospina-Giraldo, M. D., & Royse, D. J. (1999). A polymerase chain reaction-based test for the identification of Trichoderma harzianum biotypes 2 and 4, responsible for the worldwide green mold epidemic in cultivated Agaricus bisporus. Applied Microbiology and Biotechnology, 52(2), 246–250.
Esbensen, K. H., & Heydorn, K. (2004). Metrology in sampling, a first foray (biological materials). Chemometrics and Intelligent Laboratory Systems, 74(1), 115–120.
Fletcher, J. T. (1997). Mushroom spawn and the development of Trichoderma harzianum compost mold. Mushroom News, 45(8), 6–8.
Fletcher, J. T., & Gaze, R. H. (2008). Mushroom Pest and disease control. London, UK: Manson Publishing.
Garthright, W. E., & Blodgett, R. J. (2003). FDAs preferred MPN methods for standard, large or unusual tests, with a spreadsheet. Food Microbiology, 20, 439–445.
Grogan, H. M., Noble, R., Gaze, R. H., & Fletcher, J. T. (1996). Control of Trichoderma harzianum - a weed mould of the cultivated mushroom. Brighton Crop Protection - Pests & Diseases, 4(B-3), 337–343.
Halvorson, H. O., & Ziegler, N. R. (1933). Application of statistics to problems in bacteriology I. A means of determining bacterial population by the dilution method. Journal of Bacteriology, 25(2), 101–121.
Herigstad, B., Hamilton, M., & Heersink, J. (2001). How to optimize the drop plate method for enumerating bacteria. Journal of Microbiological Methods, 44(2), 121–129.
Kaufmann, D. D., Williams, L. E., & Summers, C. B. (1963). Effect of plating medium and incubation temperature on growth of fungi in soil-dilution plates. Canadian Journal of Microbiology, 9(6), 741–751.
Lane, C., Nixon, T., Woodhall, J., Glover, R., O’Brien, M. & Grogan, H. (2010). Trichoderma green mould – Diagnostic assays for improved disease management. 20p. HDC Report M48. Agriculture and Horticulture Development Board, Stoneleigh Park, Kenilworth, CV8 2TL, UK.
Largeteau, M. L., Latapy, C., Minvielle, N., Regnault-Roger, C., & Savoie, J.-M. (2010). Microbially induced diseases of Agaricus bisporus: biochemical mechanisms and impact on commercial mushroom production. Applied and Environmental Microbiology, 158, 69–83.
Lemmers, G. (2010). Trichoderma in bulk phase III (part 1). Mushroom Business, 40, 10–13.
Mamoun, M. L., Savoie, J.-M., & Olivier, J.-M. (2000b). Interactions between the pathogen Trichoderma harzianum Th2 and Agaricus bisporus in mushroom compost. Mycologia, 92, 233–240.
Mills, P. (1996). Development of a rapid diagnostic test for colonising forms of Trichoderma. 22p. HDC Report M 10a. Agriculture and Horticulture Development Board, Stoneleigh Park, Kenilworth, CV8 2TL, UK.
Mumpuni, A., Sharma, H. S. S., & Brown, A. E. (1998). Effect of metabolites produced by Trichoderma harzianum biotypes and Agaricus bisporus on their respective growth radii in culture. Applied and Environmental Microbiology, 64(12), 5053–5056.
Petersen, L., Dahl, C. K., & Esbensen, K. H. (2004). Representative mass reduction in sampling: a critical survey of techniques and hardware. Chemometrics and Intelligent Laboratory Systems, 74(1), 95–114.
Petersen, L., Minkkinen, P., & Esbensen, K. H. (2005). Representative sampling for reliable data analysis: theory of sampling. Chemometrics and Intelligent Laboratory Systems, 77(1–2), 261–277.
Rinker, D.L. (1997). Sensitivity of spawn-run compost or casing to green mould infection. Mushroom World, 44–47
Rinker, D. L., & Alm, G. (2000). Management of green mould disease in Canada. Mushroom Science, 15(2), 617–623.
Rinker, D. L., & Alm, G. (2008). Management of casing Trichoderma using fungicides. Mushroom Science, 17, 496–509.
Romaine, C. P., Royse, D. J., & Schlagnhaufer, C. (2008). Emergence of benzimidazole-resistant green mould, Trichoderma aggressivum, on cultivated Agaricus bisporus in North America. Mushroom Science, 17, 510–523.
Samuels, G. J., Dodd, S. L., Gams, W., Castlebury, L. A., & Petrini, O. (2002). Trichoderma species associated with the green mold epidemic of commercially grown Agaricus bisporus. Mycologia, 94, 147–170.
Seaby, D. A. (1987). Infection of mushroom compost by Trichoderma species. Mushroom Journal, 179, 355–361.
Seaby, D. A. (1996). Investigation of the epidemiology of green mould disease of mushroom (Agaricus bisporus) compost caused by Trichoderma harzianum. Plant Pathology, 45, 913–923.
Acknowledgments
M. O’Brien was the recipient of a Teagasc Walsh Fellowship. Part of this work was funded by the Agriculture and Horticultural Development Board, Stoneleigh Park, Warwickshire, UK. Brian McGuinness provided technical assistance with crop trials and RT-PCR.
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O’Brien, M., Kavanagh, K. & Grogan, H. Detection of Trichoderma aggressivum in bulk phase III substrate and the effect of T. aggressivum inoculum, supplementation and substrate-mixing on Agaricus bisporus yields . Eur J Plant Pathol 147, 199–209 (2017). https://doi.org/10.1007/s10658-016-0992-9
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DOI: https://doi.org/10.1007/s10658-016-0992-9