Abstract
Commercial products based on Trichoderma are obtained mainly from solid-state fermentation. Submerged liquid fermentation is the most appropriate method compared to the solid medium for large-scale production of Trichoderma spp. The present study aimed to optimize the combination of key variables that influence the liquid fermentation process of Trichoderma asperelloides LQC-96 for conidial production coupled with its efficiency in the control of Sclerotinia sclerotiorum. In addition, we verified whether the optimized culture conditions can be used for the conidial production of Trichoderma erinaceum T-12 and T-18 and Trichoderma harzianum T-15. Fermentation studies were performed in shake flasks following a planned experimental design to reduce the number of tests and consumable costs. The effect of temperature, pH, photoperiod, carbon:nitrogen ratio and water activity on conidial production were assessed, which of pH was the only meaningful factor contributing to increased conidial production of T. asperelloides LQC-96. From the five variables studied initially, pH and C:N ratio were further used in the second design (rotational central composite design—RCCD). Hence, the best conditions for the production of T. asperelloides LQC-96 conidia by liquid fermentation consisted of initial pH of 3.5, C:N ratio of 200:1 at 30 °C, without glycerol, and under 24 h photoperiod. The highest conidial concentration was observed after seven days of fermentation. Under these optimal conditions, T. erinaceum T-12 and T-18, and T. harzianum T-15 were also cultivated, but only LQC-96 efficiently parasitized S. sclerotiorum, precluding sclerotium myceliogenic germination. Our findings propose optimal fermentation conditions that maximize conidial production of T. asperelloides as a potential biofungicide against S. sclerotiorum.
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Acknowledgments
This work was financed by Embrapa Macroprograma 3 grant number 03.10.06.005.00.00. Wagner Bettiol (CNPq 303899/2015-8), Flavio HV Medeiros (CNPq 309307/2017-1), and Bernardo A. Halfeld-Vieira (CNPq 303396/2018-0) acknowledge Conselho Nacional de Desenvolvimento Científico e Tecnológico – CNPq for the productivity fellowship CNPq. ALAC, LBC, LCR, LGS, and ZVP acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – CAPES for the scholarship. Special thanks go to Dr. Ricardo Harakava, Instituto Biológico de São Paulo, for the molecular identification of Trichoderma strains.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by ALAC, LBC, LCR, LGS, and ZVP. Reagents/materials and analysis tools were obtained by BAHV, MABM, and WB. ALAC, LBC, LCR, LGS, FHVM, GMM, ZVP, and WB analyzed the data. The first draft of the manuscript was written by LCR and WB and all authors commented on previous versions of the manuscript. WB, GMM, BAHV, and FHVM, prepared the final version of the manuscript. All authors read and approved the final manuscript.
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de Rezende, L.C., de Andrade Carvalho, A.L., Costa, L.B. et al. Optimizing mass production of Trichoderma asperelloides by submerged liquid fermentation and its antagonism against Sclerotinia sclerotiorum. World J Microbiol Biotechnol 36, 113 (2020). https://doi.org/10.1007/s11274-020-02882-7
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DOI: https://doi.org/10.1007/s11274-020-02882-7