APS Abstracts of Presentations

Agrobacterium-mediated transformation of Phoma medicaginis. M. DHULIPALA, J. N. Enis, and S. M. Marek. Dept. of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078. Phytopathology 94:S25. Publication no. P-2004-0165-AMA.

Phoma medicaginis causes spring black stem and leaf spot of alfalfa and can be an important foliar pathogen of many other legumes. The objective of this study was to assess the genomic tractability of P. medicaginis as a fungal pathosystem for the model legume, Medicago truncatula. Conidia from 3 isolates of P. medicaginis from alfalfa were successfully transformed to co-express green fluorescent protein (GFP) and resistance to hygromycin B (hph) using Agrobacterium tumefaciens LBA 4404 carrying the binary vector, pPTGFPH. The conidia and hyphae of hygromycin-resistant transformants fluoresced brightly and facilitated live imaging of the infection of M. truncatula leaf cells. T-DNA integration into genome of P. medicaginis was confirmed by PCR. Analysis of the transgene insertion frequency and the genomic integration sites of transformants will be presented.

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