Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum, the maize leaf-blight fungus.
Agrobacterium tumefaciens has the ability to transfer its T-DNA to plants, yeast, filamentous fungi, and human cells and integrate it into their genome. Conidia of the maize pathogen Helminthosporium turcicum were transformed to hygromycin B resistance by a Agrobacterium- tumefaciens-mediated transformation system using a binary plasmid vector containing the hygromycin B phosphotransferase ( hph) and the enhanced green fluorescent protein ( EGFP) genes controlled by the gpd promoter from Agaricus bisporus and the CaMV 35S terminator. Agrobacterium- tumefaciens-mediated transformation yielded stable transformants capable of growing on increased concentrations of hygromycin B. The presence of hph in the transformants was confirmed by PCR, and integration of the T-DNA at random sites in the genome was demonstrated by Southern blot analysis. Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum provides an opportunity for advancing studies of the molecular genetics of the fungus and of the molecular basis of its pathogenicity on maize.
Degefu
Yeshitila
Y
Department of Applied Biology, University of Helsinki, Plant and Forest Pathology, P.O. Box 27, 00014, Finland. yeshitila.degefu@helsinki.fi
Hanif
Mubashir
M
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism; D015139:Blotting, Southern; D004269:DNA, Bacterial; D006375:Helminthosporium / Q000235:genetics*; D006921:Hygromycin B / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic; D003313:Zea mays / Q000382:microbiology*
D004269:DNA, Bacterial; C036483:T-DNA; D006921:Hygromycin B