Open AccessUse of REMI and Agrobacterium ‐mediated transformation to identify pathogenicity mutants of the biocontrol fungus, Coniothyrium minitans
Author(s) -
Rogers Christopher W.,
Challen Michael P.,
Green Jonathan R.,
Whipps John M.
Publication year - 2004
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 - 10.1016/j.femsle.2004.10.022
Subject(s) - fungus , biology , transformation (genetics) , biological pest control , mutant , pathogenicity , microbiology and biotechnology , agrobacterium , botany , gene , genetics
Restriction enzyme mediated integration (REMI) and Agrobacterium ‐mediated transformation (ATMT) were used to transform protoplasts or germinated conidia of the mycoparasite Coniothyrium minitans to hygromycin resistance. Using REMI, up to 32 transformants μg DNA −1 were obtained, while 37.8 transformants 5 × 10 5 germlings −1 were obtained using ATMT. Single‐copy integrations occurred in 8% and 40% of REMI and ATMT transformants, respectively. A novel microtitre plate‐based test was developed to expedite screening of 4000 REMI and ATMT C. minitans transformants. Nine pathogenicity mutants that displayed reduced or no pathogenicity on sclerotia of Sclerotinia sclerotiorum were identified.