Angiosperm Flora of India

Agrobacterium-mediated transformation of the barley pathogen Ramularia collo-cygni with fluorescent marker tags and live tissue imaging of infection development

Publication Type:Journal Article
Year of Publication:2011
Authors:Thirugnanasambandam, A, Wright, KM, Havis, N, Whisson, SC, NEWTON, AC
Journal:Plant Pathology
Date Published:2011
ISBN Number:1365-3059
Keywords:barley, fluorescent protein, Hordeum, Hordeum vulgare, ramularia leaf spot, transformation

Ramularia collo-cygni, the causal agent of ramularia leaf spot disease in barley was transformed to express the green fluorescent protein and DsRed fluorescent protein using a combined protoplast and Agrobacterium tumefaciens-mediated transformation technique. Regenerated protoplasts from mycelial fragments were transformed with A. tumefaciens strain AGL1 containing either pCAMBgfp or pCAMBDsRed plasmids. Two different isolates of R. collo-cygni were transformed and several stable transformants were recovered. These transformants infected detached barley leaves and characteristic swan’s-neck-shaped conidiophores were seen on detached leaves 9 days after inoculation, as well as spore-like structures formed from hyphal swellings. This first successful transformation of R. collo-cygni has enabled better description of the stages of disease development and pathogen behaviour during the endophytic/asymptomatic stage of its life cycle.

Short Title:Plant Pathology
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