Transformation of Phomopsis viticola with the green fluorescent protein

Abstract

Phomopsis viticola is the causal agent of Phomopsis cane and leaf spot on Vitis spp., a persistent 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 mitotically stable transformants with no change in virulence on grape internodes and leaves in comparison to the wild type. These transformants will be critical tools for elucidating fungal penetration of host plants, invasive growth and the nature of its host association.

• cane and leaf spot
• pBHt2_sGFP
• pIGPAPA
• protoplast
• Vitis

Transformations were made possible with the technical guidance of Dr Soonok Kim of Ohio State University. We are also grateful for the contributions of Mr Lee Wilson and the staff at the Molecular and Cellular Imaging Center at the OARDC. We additionally thank Dr Stephen Marek and Dr Nathan Walker at Oklahoma State University for the pBHt2_sGFP vector and Dr Lynda Chiuffetti at Oregon State University for providing the ToxA promoter. Financial support for this project was provided by the Viticulture East Consortium and the Ohio Grape Industries Program.