Compositions and methods for the isolation and purification of anti-fungal protein from epichloe festucae and use thereof for reducing symptoms of dollar spot disease in targeted plant species
AssigneeRutgers, The State University of New Jersey
InventorsFaith C. BELANGER, Bruce B. CLARKE, Patrick FARDELLA
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Abstract
Compositions and methods are provided for prevention and treatment of plant diseases with an antifungal protein.
Claims29
A method for increasing yield of Efe-AfpA antifungal protein from transgenic P. chrysogenumEfe-AfpA comprising;
- a) culturing conidia from P. chrysogenumEfe-AfpA in high nutrient media for a suitable time period to produce mycelia without inducing Ef-AfpA expression;
- b) harvesting and culturing the mycelia of step a) in a minimal low nutrient media for a suitable time period to induce robust Efe-AfpA expression and secretion into said low nutrient media,
- c) centrifuging and filtering said low nutrient media to remove conida and excess debris;
- d) dialyzing said filtered media of step c) on an ion exchange column;
- e) eluting said Efe-AfpA with increasing salt concentrations to form an eluant;
- and
- f) filtering and desalting said eluant via passage through filter, thereby isolating said Efe-AfpA antifungal protein.
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A method for increasing yield of Efe-AfpA antifungal protein from transgenic Penicillium sp transduced with a nucleic acid encoding Efe-AfpA;
- a) culturing conidia from said transduced Penicillium sp. in high nutrient media for a suitable time period to produce mycelia without inducing Efe-AfpA expression;
- b) harvesting and culturing the mycelia of step a) in a minimal low nutrient media for a suitable time period to induce robust Efe-AfpA expression and secretion into said low nutrient media,
- c) centrifuging and filtering said low nutrient media to remove conida and excess debris;
- d) dialyzing said filtered media of step c) on an ion exchange column;
- e) eluting said Efe-AfpA with increasing salt concentrations to form an eluant;
- and
- f) filtering and desalting said eluant via passage through filter, thereby isolating said Efe-AfpA antifungal protein.
A transgenic plant, transgenic fungus, or transgenic bacteria comprising an antifungal protein from Epichloe festucae, E. gansuensis var. inebrians. E. baconii, E. aotearoae, and E. coenophiala.
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Dependencies are inferred from the claim text.