APS Abstracts of Presentations

Transformation of Botrytis cinerea with GFP - A tool to study the cytology of silicon-induced resistance against gray mold
S. KAMENIDOU (1), S. Casanova (1), M. R. Dhulipala (1), J. N. Enis (1), T. J. Cavins (2), S. M. Marek (1)
(1) Oklahoma State University; (2) Sun Gro Horticulture
Phytopathology 96:S57

Botrytis cinerea Pers. causes gray mold and Botrytis blight, two of the most common and economically important diseases of greenhouse floricultural crops. Greenhouse crops grown in soilless peat-mixes are deficient in silicon (Si). Cell wall silica accumulation results in tissue fortification, creating a physical barrier thought to be responsible for plant disease resistance. In preliminary studies, Si supplementation of sunflowers reduced Botrytis blight stem lesion incidence and severity. In order to better study this host-pathogen interaction, conidia of B. cinerea strain Gerbera-1 were transformed using Agrobacterium tumefaciens AGL-1 carrying one of three different binary vectors: pBHt2-sGFP, pOHT-sGFP or pOHT-DsRed. Hygromycin resistance (hph) was under the control of either the trpC or oliC promoter. Green fluorescent protein (sGFP) expression was driven by the ToxA promoter and DsRed fluorescent protein was driven by the gpdA promoter. Transformants were purified by subculturing single-conidia twice and all expressed resistance to 100 µg/ml hygromycin and variable green or red fluorescence. T-DNA integration was confirmed by PCR. The brightest green fluorescent transformants were selected to study the cytology of Si-induced resistance against B. cinerea.

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