Chan, John , Perry, Greg , Pauls, K. Peter .
An Agrobacterium-infiltration transient expression assay adopted for Phaseolus vulgaris L.
Agrobacterium-mediated transient expression has been used by investigators as a convenient means of assaying the effects of transgenes in a variety of plant models like Nicotiana tabacum and Arabidopsis thaliana. We have modified an Agroinfiltration protocol first reported by Behamane et al. 2000 to accommodate transient gene expression assays in Phaseolus vulgaris. The A. tumefaciens strain GV3101 was transformed with various BiBAC2 constructs containing both GFP under the control of an enhanced CaMV 35S constitutive promoter and GUS-NPTII reporter genes or GUS-NPTII reporter gene alone and P. vulgaris BAC inserts of 150 kb and 300 kb respectively. Reporter gene expression was analyzed and patterns of successful transient expression were compared for the different constructs using vacuum infiltration on young seedlings and individual leaves on older plants. We have developed this transient expression assay as a method to conveniently screen for genes conferring resistance to common bacterial blight (CBB) in a BiBAC2 expression library constructed from the CBB-resistant P. vulgaris L. variety OAC Rex.
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1 - University of Guelph, Plant Agriculture, 5 Stone Rd., Guelph, Ontario, N1G 2W1, Canada
2 - University of Guelph, Plant Agriculture
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM