Production of Conidia by the Fungus Metarhizium anisopliae Using Solid-State Fermentation

Methods Mol Biol. 2016:1477:61-9. doi: 10.1007/978-1-4939-6367-6_6.

Abstract

This chapter describes the production of conidia by Metarhizium anisopliae using solid-state fermentation. Before production of conidia, procedures for strains conservation, reactivation, and propagation are essential in order to provide genetic stability of the strains. The strain is conserved in freeze-dried vials and then reactivated through insect inoculation. Rice is used as a substrate for the conidia production in two different bioreactors: plastic bags and tubular bioreactor. The CO2 production in the tubular bioreactors is measured with a respirometer; this system allows calculating indirect growth parameters as lag time (tlag) (25-35 h), maximum rate of CO2 production (rCO2 max) (0.5-0.7 mg/gdm h), specific rate of CO2 production (μ) (0.10-0.15 1/h), and final CO2 production (CO2) (100-120 mg/gdm). Conidial yield per gram of dry substrate (gdm) should be above 1 × 10(9) conidia/gdm after 10 days of incubation. Germination and viability of conidia obtained after 10 days of incubation should be above 80 % and 75 %, respectively. Bioassays using of Tenebrio molitor as a host insect should yield a final mortality above 80 %.

Keywords: Biological control; Entomopathogenic fungi; Metarhizium anisopliae; Solid-state fermentation; Tenebrio molitor.

MeSH terms

  • Biological Control Agents*
  • Bioreactors
  • Culture Media
  • Fermentation*
  • Metarhizium / growth & development*
  • Spores, Fungal*

Substances

  • Biological Control Agents
  • Culture Media