Open AccessTransformation of the entomopathogenic fungus Metarhizium anisopliae to benomyl resistance
Author(s) -
Bernier L.,
Cooper R.M.,
Charnley A.K.,
Clarkson J.M.
Publication year - 1989
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1989.tb03483.x
Subject(s) - biology , benomyl , metarhizium anisopliae , entomopathogenic fungus , microbiology and biotechnology , metarhizium , transformation (genetics) , botany , gene , conidium , genetics , biological pest control , beauveria bassiana , fungicide
Protoplasts of the entomopathogenic fungus Metarhizium anisopliae were transformed to benomyl resistance using cosmid pSV50 which harbours a β‐tubulin gene cloned from a Neurospora crassa benomyl‐resistant mutant. Transformant colonies, which appeared at a frequency of 4 per 50 μg DNA, grew and sporulated on 10 μg/ml benomyl, whereas the wild type was inhibited by 3 μg/ml. Southern blot hybridization of DNA from transformants showed that, in each case, tandem repeats of the cosmid had integrated at several chromosomal loci. The transformants were mitotically stable when subcultured on non‐selective agar and retained the ability to infect and kill larvae of Manduca sexta . Two transformants were less virulent than the wild type and one of them showed slower in vitro spore germination. The benomyl‐resistant phenotype persisted in reisolates from insect cadavers.