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United States Department of Agriculture

Agricultural Research Service

Title: Isolation of a Pathogenesis Related Gene (Pr Gene) Using Laser Capture Microdissection of Glycine Max (Soybean) Syncytia

Authors
item Klink, Vincent
item Macdonald, Margaret
item Matthews, Benjamin

Submitted to: BARC Poster Day
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 2006
Publication Date: May 1, 2006
Citation: Klink, V.P., Macdonald, M.H., Matthews, B.F. 2006. Isolation of a pathogenesis related gene (pr gene) using laser capture microdissection of glycine max (soybean) syncytia [abstract]. BARC Poster Day. Paper No. 24.

Technical Abstract: The soybean cyst nematode (SCN), Heterodera glycines Ichinohe, is the major pest of soybean, Glycine max, and causes an estimated half to one billion dollars in losses annually in the U.S. Our goal is to broaden resistance of soybean to the multiple races of SCN using biotechnology. To identify genes expressed at the feeding site of SCN in soybean, roots of soybean, cv. Kent L. Merr., plants susceptible to the soybean cyst nematode (SCN), were inoculated and allowed to develop feeding sites (syncytia) for eight days. Root samples enriched in syncytial cells were collected using laser capture microdissection (LCM). RNA was extracted and used to make a cDNA library and expressed sequence tags (ESTs) were produced and analyzed. A pathogenesis-related (PR) gene was identified by these EST analyses. RT-PCR results indicated enhanced expression of various genes, including the PR gene, in syncytium samples as compared to samples extracted from whole roots. To analyze the function of these genes expressed in the syncytium, we developed a rapid-cycling soybean variety for transformation studies. MiniMax is shown to be suitable for hairy-root transformation and produce somatic embryos, thus making them a useful alternative for transformation studies and studying the pathogenicity of SCN in soybean. These data and tools will help scientists develop soybean with broader resistance to SCN.

Last Modified: 7/25/2014
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