APS Abstracts of Presentations
Agrobacterium tumefaciens-mediated transformation of Monilinia
fructicola with green fluorescent protein. S. M. MAREK (1), Z. Pan
(1), L. M. Ciuffetti (2), and R. M. Bostock (1). (1) Dept. Plant Pathology,
University of California, Davis, CA 95616; (2) Dept. Botany & Plant
Pathology, Oregon State University, Corvalis, OR 97331. Publication no.
P-2002-0673-AMA.
Monilinia fructicola causes brown rot of many stone fruits. In order to
genetically manipulate this fungus, a transformation system using Agrobacterium
was developed. The binary transformation vector, pPTGFPH, was constructed
from the plasmid pCT74, carrying green fluorescent protein (GFP) driven by the
ToxA promoter of Pyrenophora tritici-repentis and hygromycin B
phosphotransferase (hph) under control of the trpC promoter of
from Aspergillus nidulans, and the binary vector pCB403.2, carrying
neomycin phosphotransferase (nptII) between the T-DNA borders.
Macroconidia of M. fructicola were co-incubated with A. tumefaciens strain
LBA 4404(pPTGFPH) on media containing acetosyringone for two days. Hygromycin-
and G418-resistant M. fructicola transformants were selected while
inhibiting A. tumefaciens with cefotaxime. Transformants expressing GFP
fluoresced brightly. Transformation efficiency, T-DNA integration frequency,
and in situ studies on stone fruit tissues will be presented.
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