Open AccessAgrobacterium tumefaciens ‐mediated transformation of Leptosphaeria spp. and Oculimacula spp. with the reef coral gene DsRed and the jellyfish gene gfp
Author(s) -
Eckert Maria,
Maguire Kerry,
Urban Martin,
Foster Simon,
Fitt Bruce,
Lucas John,
HammondKosack Kim
Publication year - 2005
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.2005.09.041
Subject(s) - leptosphaeria maculans , biology , agrobacterium tumefaciens , transformation (genetics) , agrobacterium , mycelium , reporter gene , botany , gene , microbiology and biotechnology , genetics , gene expression
Four filamentous ascomycetes, Leptosphaeria maculans , L. biglobosa , Oculimacula yallundae and O. acuformis , were transformed via Agrobacterium tumefaciens ‐mediated transformation with the genes encoding DsRed and GFP. Using vectors pCAMDsRed and pCAMBgfp, either germinated conidia of Leptosphaeria spp. and O. yallundae or physically fragmented cultures of Oculimacula spp. were transformed. In vitro, the expression of the two reporter proteins in mycelium of both Oculimacula and both Leptosphaeria species was sufficient to distinguish each species in co‐inoculated cultures. In planta , transformants of L. maculans or L. biglobosa expressing DsRed or GFP could be observed together in leaves of Brassica napus. Either reporter protein could be used to view the colonization of leaf petioles by both Leptosphaeria spp. and growth in the xylem vessels could be clearly observed. With the generation of these transformants, further studies on interactions between pathogen species involved in disease complexes on various host species and between opposite mating types of the same species are now possible.