Submitted to: Mid Atlantic Plant Molecular Biology Society Conference
Publication Type: Abstract Only
Publication Acceptance Date: 9/1/2005
Publication Date: 9/1/2005
Citation: Martins, V., Klink, V.P., Alkharouf, N., Macdonald, M.H., Matthews, B.F. 2005. Isolation and characterization of the soybean cyst nematode (heterodera glycines) homolog of caenorhabditis elegans unc-97 gene [abstract]. Mid Atlantic Plant Molecular Biology Society Conference. p. 13. .
Technical Abstract: The soybean cyst nematode (SCN) is the major pest of soybean in the United States and no agronomically important soybean cultivars are resistant to all races of SCN. Our major goal is to develop new modes of soybean resistance to SCN using biotechnology. Therefore, we identified and characterized the SCN (Heterodera glycines) homolog of the Caenorhabditis elegans unc-97 gene (Hg-unc97) a gene that may be essential for SCN survival. In C. elegans the unc-97 gene encodes a protein involved in movement that constitutes a component of the muscular adherens junction by interacting with the beta-integrin PAT-3 at the focal adhesion-like attachment sites of muscles. In C. elegans mutants of unc-97 are limp, egg laying-defective and paralyzed. We identified a SCN partial cDNA clone, Hg-unc97, with high amino acid identity to C. elegans unc-97 and used PCR to obtain the full-length Hg-unc97 gene. The Hg-unc-97 cDNA is 1574 nt, encoding a 522 aa protein. Phylogenetic analysis of the full-length sequence revealed 63% amino acid identity with C. elegans unc-97. Real-time reverse transcription polymerase chain reaction (RT-PCR) indicated that Hg-unc-97 is expressed in all stages of the SCN life cycle. The relative ratios of Hg-unc97 between the different time points in the life cycle obtained by RT-PCR indicated a 6-fold increase in Hg-unc-97 RNA in J2s, respective to eggs, and nematodes 15 and 30-day post infection. The use of the Hg-unc-97 to broaden resistance of soybean to SCN will be tested in future experiments.