Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: December 1, 2005
Publication Date: December 1, 2005
Citation: Klink, V.P., Macdonald, M.H., Alkharouf, N., Matthews, B.F. 2005. Isolation of genes using laser capture microdissection of glycine max (soybean) syncytia formed by the plant pathogen heterodera glycines, (soybean cyst nematode) [abstract]. Cambridge Healthtech Institute Laser Capture Microdissection: Cutting Edge Applications. Paper No. 219.
Roots of soybean, Glycine max cv. Kent L. Merr., plants susceptible to the soybean cyst nematode (SCN), Heterodera glycines Ichinohe, 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. The cDNAs encoding unique '-tubulin (GmTubA1), '-tubulin (GmTubB4), and an aquaporin (GmPIP2,2) were identified by EST analyses. Immunolocalization and image analyses within and around the syncytium indicated increased levels of '-tubulin within the syncytium. However, '-tubulin labeling appeared diffuse or clumped. RT-PCR results indicated enhanced expression of GmTubA1, GmTubB4, GmPIP2,2, and several other genes including a pathogen resistance gene in syncytium-enriched samples as compared to samples extracted from whole roots. The isolation of these cDNAs has provided a bank of genes we are using for both overexpression and RNAi studies.