Culture conditions affect virulence and production of insect toxic proteins in the entomopathogenic fungus Metarhizium anisopliae.

Conidial spores are often used as the infectious agent during insect biocontrol applications of entomopathogenic fungi. Here we show differential virulence of conidia derived fromMetarhizium anisopliaestrain EAMa 01/58-Su depending upon the solid substrata used for cultivation, where LC50values differed by up to ~10-fold (5.3×106−4.5×105conidia/ml) and LT50values by ~40% (9.8−7.1 d). This fungal strain is also known to secrete proteins that are toxic towards adult Mediterranean fruit flies,Ceratitis capitata, and the Greater wax moth,Galleria mellonella, larvae.In vitroproduction and intrahemoceol injection usingG. mellonellaas the host was used to test fractions during purification of the protein toxins, demonstrating that they elicited defence-related responses including melanisation and tissue necrosis. Production of these proteins/peptides along with a number of potential cuticle degrading enzymes was confirmed bothin vitroand during the infection process (in vivo). Two-dimensional gel electrophoresis, followed by gel elution and bioassay, was used to identify at least three proteins or peptides (molecular mass=11, 15 and 15 kDa) as mediating the observed insect toxicity. These data demonstrate thatin vitroscreening for insect toxins can mimicin vivo(i.e. during the infection process) secretion and applies the use of proteomics to invertebrate pathology. [ABSTRACT FROM AUTHOR]