Agrobacterium tumefaciens-mediated transformation of Leptosphaeria spp. and Oculimacula spp. with the reef coral gene DsRed and the jellyfish gene gfp.
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.
Eckert
Maria
M
Rothamsted Research, Harpenden, Herts, AL5 2JQ, UK.
Maguire
Kerry
K
Urban
Martin
M
Foster
Simon
S
Fitt
Bruce
B
Lucas
John
J
Hammond-Kosack
Kim
K
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000818:Animals; D037421:Anthozoa / Q000235:genetics; D001203:Ascomycota / Q000235:genetics* / Q000472:pathogenicity; D001483:Base Sequence; D029688:Brassica napus / Q000382:microbiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D010935:Plant Diseases / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D037401:Scyphozoa / Q000235:genetics; D014170:Transformation, Genetic
D008164:Luminescent Proteins; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins