Publication Type: | Journal Article |
Year of Publication: | 2011 |
Authors: | Thirugnanasambandam, A, Wright, KM, Havis, N, Whisson, SC, NEWTON, AC |
Journal: | Plant Pathology |
Volume: | 60 |
Issue: | 5 |
Date Published: | 2011 |
ISBN Number: | 1365-3059 |
Keywords: | barley, fluorescent protein, Hordeum, Hordeum vulgare, ramularia leaf spot, transformation |
Abstract: | 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. |
URL: | http://dx.doi.org/10.1111/j.1365-3059.2011.02440.x |
Short Title: | Plant Pathology |
Agrobacterium-mediated transformation of the barley pathogen Ramularia collo-cygni with fluorescent marker tags and live tissue imaging of infection development
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