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United States Department of Agriculture

Agricultural Research Service

Title: Laser Capture Microscopy Isolation of Syncytial Cells from Heterodera Glycines Feeding Sites and Est Analysis

item Klink, Vincent
item Macdonald, Margaret
item Alkharouf, Nadmi - GEORGE MASON UNIVERSITY
item Matthews, Benjamin

Submitted to: Meeting Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: February 4, 2004
Publication Date: February 4, 2004
Citation: Klink, V.P., Macdonald, M.H., Alkharouf, N., Matthews, B.F. 2004. Laser capture microscopy isolation of syncytial cells from heterodera glycines feeding sites and est analysis. Meeting Proceedings. Cambridge Healthtech Institute. P.6.

Technical Abstract: Heterodera glycines, the soybean cyst nematode (SCN), is the major pathogen of soybeans comprising approximately one billion dollars in losses, annually. SCN causes their damage by establishing a feeding site near the root vasculature. As hundreds of nematodes invade and establish feeding sites, the plant can eventually weaken and die. We used laser capture microscopy (LCM) as a means to retrieve mRNAs that are expressed within the developing syncytia of Glycine max cv. Kent. (soybeans). Over 500 cDNAs were isolated from a library generated from 8-day old microdissected syncytia. These clones could be further condensed into 196 unique contigs. While some of the cDNAs have bona fide matches to sequences in the database with known function, many of the sequences are of unknown function. One cDNA encodes an extensin, a cell wall protein known to be involved in defense responses. This extensin (soybean hydroxyproline rich protein #4 [SBHGRP4]) is a new member of the extensin gene cluster. A second gene, GMIPER, a peroxidase, is a gene that is known to specifically cross-link extensin in other plants. Both of these genes have been implicated in host defense responses. These data provide evidence that mRNA can be extracted from developing syncytial cells. Some of these mRNAs have the potential to be used to control SCN.

Last Modified: 3/27/2015
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