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Production of microsclerotia by Brazilian strains of Metarhizium spp. using submerged liquid culture fermentation

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Abstract

We investigated the potential production and desiccation tolerance of microsclerotia (MS) by Brazilian strains of Metarhizium anisopliae (Ma), M. acridum (Mc) and M. robertsii (Mr). These fungi were grown in a liquid medium containing 16 g carbon l−1 with a carbon:nitrogen ratio of 50:1. One hundred milliliters cultures were grown in 250 ml Erlenmeyer flasks in a rotary incubator shaker at 28 °C and 200 rpm for 5 days. Five-day-old MS were harvested, mixed with diatomaceous earth (DE) and air-dried for 2 days at 30 °C. The air-dried MS–DE granular preparations were milled by mortar + pestle and stored in centrifuged tubes at either 26 or −20 °C. Desiccation tolerance and conidia production were assessed for dried MS granules by measuring hyphal germination after incubation for 2 days on water agar plates at 26 °C and for conidia production following 7 days incubation. Yields of MS by all strains of Metarhizium were 6.1–7.3 × 106 l−1 after 3 days growth with maximum MS yields (0.7–1.1 × 107 l−1) after 5 days growth. No differences in biomass accumulation were observed after 3 days growth, whereas Ma-CG168 showed the highest biomass accumulation after 5 days growth. Dried MS–DE preparations of all fungal strains were equally tolerant to desiccation (≥93 % germination) and the highest conidia production was obtained by MS granules of Mc-CG423 (4 × 109 conidia g−1). All MS granules showed similar stability after storage at either 26 or −20 °C for 3.5 months.

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Acknowledgments

This work was funded by Embrapa (Empresa de Pesquisa Brasileria de Agropecuária, Brasília, Brazil). We thank Rogério B. Lopes and Marcos R. Faria (Embrapa Cenargen) for providing the strains of Metarhizium spp. and for measuring water activities.

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Authors and Affiliations

  1. Embrapa Rice and Beans, Rodovia GO-462, Km 12, Zona Rural, C.P. 179, Santo Antônio de Goiás, GO, 75375-000, Brazil

    Gabriel Moura Mascarin, Nilce Naomi Kobori, Rayan Carlos de Jesus Vital & Eliane Dias Quintela

  2. Crop Bioprotection Research Unit, United States Department of Agriculture, Agriculture Research Service, National Center for Agricultural Utilization Research, 1815 N. University St, Peoria, IL, 61604, USA

    Mark Alan Jackson

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  1. Gabriel Moura Mascarin
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  2. Nilce Naomi Kobori
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  3. Rayan Carlos de Jesus Vital
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  4. Mark Alan Jackson
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  5. Eliane Dias Quintela
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Correspondence to Gabriel Moura Mascarin.

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Mascarin, G.M., Kobori, N.N., de Jesus Vital, R.C. et al. Production of microsclerotia by Brazilian strains of Metarhizium spp. using submerged liquid culture fermentation. World J Microbiol Biotechnol 30, 1583–1590 (2014). https://doi.org/10.1007/s11274-013-1581-0

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