In a previous study, a spontaneous subtilisin pr1A and pr1B gene-deficient mutant of the entomopathogenic fungus Metarhizium anisopliae strain V275 has been identified [Wang, C.-S. et al. (2002) FEMS Microbiol. Lett. 213, 251^255]. The insecticidal metabolites of this mutant were studied further. High-performance liquid chromatography (HPLC) analysis indicated that the mutant isolate lost the ability to produce cyclic peptide toxins, destruxins, both in vitro and in vivo. Pulsed-field gel electrophoresis revealed that the mutant concurrently lost a 1.05 Mb (approximately) chromosome, demonstrating for the first time that a conditionally dispensable (CD) chromosome exists in the insect pathogenic fungus, M. anisopliae. Concurrence of losing the ability to produce destruxins and a CD chromosome in the mutant suggests that the toxin synthetase genes of M. anisopliae are located on this CD chromosome, as similarly described for plant pathogenic fungi. Semi-quantitative api ZYM analysis showed more biochemical disparities between the mutant and the wild-type strain. ß
There is much public interest in the use of fungal biological control agents as alternatives to chemical pesticides. However, there are some concerns as to whether the metabolites produced by these fungi pose a risk to humans and the environment. Destruxins are the main metabolites produced by the insect pathogenic fungus Metarhizium anisopliae (Metsch.) Sorok. The production of these compounds in two different insect hosts and their subsequent fate in the soil were assessed as a case study. Destruxin profiling revealed that the amount and type of destruxin produced was dependant upon the fungal strain and insect host and that these compounds decomposed shortly after host death. Destruxin decomposition was presumably due to the activity of hydrolytic enzymes in the cadavers and appeared to be independent of host or soil type and biota. Temperature strongly influenced destruxin decomposition. Our studies are the first to show that the destruxins are essentially restricted to the host and pathogen and are unlikely to contaminate the environment or enter the food chain.
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