Open AccessCo‐transformation of Metarhizium anisopliae by electroporation or using the gene gun to produce stable GUS transformants
Author(s) -
Leger Raymond J.St.,
Shimizu Susumu,
Joshi Lokesh,
Bidochka Michael J.,
Roberts Donald W.
Publication year - 1995
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.1574-6968.1995.tb07789.x
Subject(s) - metarhizium anisopliae , biology , electroporation , transformation (genetics) , plasmid , benomyl , microbiology and biotechnology , hypha , entomopathogenic fungi , hygromycin b , reporter gene , gene , botany , genetics , gene expression , fungicide , biological pest control
The potential of β‐glucuronidase as a molecular marker for studying the environmental microbiology of entomopathogenic fungi was assessed. Metarhizium anisopliae was stably co‐transformed with plasmids (pNOM102 and pBENA3) containing the β‐glucuronidase and benomyl resistance (β‐tubulin) genes, using both electroporation and biolistic delivery systems, and it was confirmed that the expressed phenotypes were not exhibited by ten randomly chosen indigenous North‐American isolates. In spite of random and multiple integrations, the co‐transformants showed normal growth rates and retained their pathogenicity to insects. β‐Glucuronidase activity in the co‐transformants was used to detect histochemically the presence of fungal hyphae in infected host insects ( Bombyx mori ) and thus provides a practical means of marking genetically engineered pathogens for field trials.