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Co-bombardment,integration and expression of rice chitinase and thaumatin-like protein genes in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> cv. Conlon)
Authors:Dennis J Tobias  Muthusamy Manoharan  Clara Pritsch  Lynn S Dahleen
Institution:(1) Department of Plant Sciences, North Dakota State University, Fargo, ND 58105, USA;(2) Department of Agriculture, University of Arkansas, Pine Bluff, AR 71601, USA;(3) Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la Repóblica, Garzón 780, Montevideo, Uruguay;(4) Cereal Crops Research Unit, USDA-ARS Northern Crop Science Lab, Fargo, ND 58105, USA
Abstract:Pathogenesis-related (PR) proteins associated with degradation of structural components of pathogenic filamentous fungi were overexpressed in the two-rowed malting barley (Hordeum vulgare L.) cultivar Conlon. Transgenes were introduced by co-bombardment with two plasmids, one carrying a rice (Oryza sativa L.) chitinase gene (chi11) and another carrying a rice thaumatin-like protein gene (tlp). Each gene was under the control of the maize ubiquitin (Ubi1) promoter. Fifty-eight primary transformants from three independent transformation events were regenerated. T1 plants with high rice chi11 and tlp protein expression levels were advanced to identify T2 homozygotes by herbicide spray and subjected to further molecular analyses. T3 progeny from one event (E2) had stable integration and expression of the rice chi11 and tlp while those from the other events (E1 and E3) showed stable integration only of tlp. The successful production of these lines overexpressing the antifungal chi and tlp proteins provides materials to test the effects of these genes on a variety of fungal diseases that attack barley and to serve as potential additional sources of disease resistance.
Keywords:Oryza sativa            PR-proteins  Transgene expression  Transgene silencing
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