Growth substrates and caleosin-mediated functions affect conidial virulence in the insect pathogenic fungus Beauveria bassiana.

The entomopathogenic fungus, Beauveria bassiana, is a microbial biological control agent capable of infecting a wide range of insect hosts. Conidia (spores) initiate infection via adhesion, growth and penetration of the insect cuticle, whose outmost layer is rich in lipids. Conidial virulence was investigated in B. bassiana WT and caleosin mutants (ΔBbcal1), the latter a protein involved in lipid storage and turnover. Topical insect bioassays revealed that conidia of the WT strain showed up to 40-fold differences in LD50 values depending upon the growth substrate. The most virulent conidia were harvested from potato dextrose agar containing oleic acid, and the least potent were those derived from Sabouraud dextrose/yeast extract agar (SDAY). However, with the exception of conidia derived from SDAY and Czapek Dox agar, in which values were reduced, mean lethal times to kill (LT50) were essentially unaffected. In topical bioassays, the ΔBbcal1 mutant displayed LD50 values 5-40-fold higher than the WT depending upon the growth substrate, with ΔBbcal1 conidia derived from SDAY unable to effectively penetrate the host cuticle. The ΔBbcal1 mutant also showed concomitant dramatic increases in LT50 values from a mean of ~4.5 for WT to >8.5 days for the mutant. In contrast, intrahaemocoel injection bioassays that bypass cuticle penetration events revealed only minor effects on virulence for either WT or ΔBbcal1 conidia. These data highlight the importance of caleosin-dependent lipid mobilization and/or signalling in cuticle penetration events but suggest their dispensability for immune evasion and within-host growth.