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Co-bombardment, integration and expression of rice chitinase and thaumatin-like protein genes in barley (Hordeum vulgare cv. Conlon)

  • Genetic Transformation and Hybridization
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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.

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Acknowledgments

This material is based upon the work supported by the U.S. Department of Agriculture. This is a cooperative project with the U.S. Wheat and Barley Scab Initiative (USWBSI 0405-DA-067). The authors are grateful to Dr. R. Skadsen, (USDA-ARS, Madison, WI) for the antiserum of tlp and Dr. S. Muthukrishnan (Kansas State University, Manhattan, KS) for the plasmids and chi antiserum. We thank W. Morgan, N. Hillen and A. Jha for technical support. The use of trade, firm, or corporation names in this publication is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable.

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Authors and Affiliations

  1. Department of Plant Sciences, North Dakota State University, Fargo, ND, 58105, USA

    Dennis J. Tobias

  2. Department of Agriculture, University of Arkansas, Pine Bluff, AR, 71601, USA

    Muthusamy Manoharan

  3. Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la Repóblica, Garzón 780, Montevideo, Uruguay

    Clara Pritsch

  4. Cereal Crops Research Unit, USDA-ARS Northern Crop Science Lab, Fargo, ND, 58105, USA

    Lynn S. Dahleen

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  1. Dennis J. Tobias
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  2. Muthusamy Manoharan
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  3. Clara Pritsch
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  4. Lynn S. Dahleen
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Correspondence to Lynn S. Dahleen.

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Communicated by P. P. Kumar

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Tobias, D.J., Manoharan, M., Pritsch, C. et al. Co-bombardment, integration and expression of rice chitinase and thaumatin-like protein genes in barley (Hordeum vulgare cv. Conlon). Plant Cell Rep 26, 631–639 (2007). https://doi.org/10.1007/s00299-006-0263-x

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  • DOI: https://doi.org/10.1007/s00299-006-0263-x

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