Transformation of Phomopsis viticola with the Green Fluorescent Protein
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Date
2009-03-31T17:50:26Z
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Abstract
Phomopsis viticola is the causal agent of Phomopsis cane and leaf spot on Vitis spp., which is a serious and economically important disease in temperate regions. Here we describe the transformation of this fungus with two different constructs (pBHt2_sGFP and pIGPAPA) containing the green fluorescent protein (GFP) and the hygromycin B resistance gene (hph). Protoplast mediated transformation yielded three fluorescent transformants (DA-2, DA-4, and DA-5) and multiple non-fluorescent transformants. All transformants obtained were mitotically stable. The growth rates of DA-4 and DA-5 were slightly, but significantly (P < 0.05), less than DA-2 and OH-22 (wild type); however, transformation resulted in no change in sporulation rate, log spore production, and virulence on grape internodes and leaves compared to the wild type. These transformants will be useful tools for elucidating fungal penetration of host plants, invasive growth, and the nature of its host association.
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Phomopsis cane, protoplast transformation, leaf spot