Transformation of Phomopsis viticola with the green fluorescent protein

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.

[1]  L. L. Wilson,et al.  Effects of Application of Fungicide During the Dormant Period on Phomopsis Cane and Leaf Spot of Grape Disease Intensity and Inoculum Production. , 2006, Plant disease.

[2]  Karl-Heinz Kogel,et al.  Infection patterns in barley and wheat spikes inoculated with wild-type and trichodiene synthase gene disrupted Fusarium graminearum. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[3]  A. Rossman,et al.  Characterization of Phomopsis spp. Infecting Grapevines in the Great Lakes Region of North America. , 2005, Plant disease.

[4]  L. Madden,et al.  Temperature and Wetness-Duration Requirements for Grape Leaf and Cane Infection by Phomopsis viticola. , 2003, Plant disease.

[5]  R. Carlucci,et al.  PHOMOPSIS VITICOLA IS EASILY TRANSFORMED WITH hph AND Bmlr GENES , 2003 .

[6]  L. Madden,et al.  Effect of Growth Stage on Susceptibility of Grape Berry and Rachis Tissues to Infection by Phomopsis viticola. , 2001, Plant disease.

[7]  M. Dickman,et al.  Green Fluorescent Protein Is Lighting Up Fungal Biology , 2001, Applied and Environmental Microbiology.

[8]  D. Geiser,et al.  Agrobacterium-Mediated Transformation of Fusarium oxysporum: An Efficient Tool for Insertional Mutagenesis and Gene Transfer. , 2001, Phytopathology.

[9]  G. May,et al.  Development of a transformation system for Mycosphaerella pathogens of banana: a tool for the study of host/pathogen interactions. , 2001, FEMS microbiology letters.

[10]  B. Horwitz,et al.  A G protein alpha subunit from Cochliobolus heterostrophus involved in mating and appressorium formation. , 1999, Fungal genetics and biology : FG & B.

[11]  Laurence V. Madden,et al.  The study of plant disease epidemics , 2007 .

[12]  I. Connerton,et al.  Characterization of the glyoxysomal isocitrate lyase genes of Aspergillus nidulans (acuD) and Neurospora crassa (acu-3) , 2004, Current Genetics.

[13]  Ömer Erincik,et al.  Studies to determine time of grape berry and rachis susceptibility, and environmental parameters required for leaf and cane infection by phomopsis viticola / , 2002 .

[14]  R. Pearson,et al.  Compendium of grape diseases. , 1989 .

[15]  T. Wicks.  Compendium of Grape Diseases , 1989, Australasian Plant Pathology.

[16]  T. Pine.  Development of the Grape dead-arm disease. , 1959 .

[17]  J. G. Horskfall.  An Improved Grading System For Measuring Plant Diseases Vol-36 , 1945 .

[18]  D. Reddick.  Dead-arm disease of grapes. , 1914 .