Submitted to: Nematology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 2, 2002
Publication Date: December 1, 2002
Citation: Masler, E. P. 2002. A polyclonal antiserum that recognizes the major female-specific proteins, vitellogenins, in the soybean cyst nematode Heterodera glycines. International Journal of Nematology 12: 119-124. Interpretive Summary: Plant-parasitic nematodes attack all crops of agricultural importance, causing over $10 billion in losses annually to U.S. farmers. Problems facing growers include environmental questions about a widely used chemical nematicide, and the continued spread of parasitic nematodes to uninfested soil. This makes the development of new environmentally and economically sound detection and control systems critical. This report describes the production and characteristics of an antibody that detects specific female proteins in the soybean cyst nematode, a serious soybean pest. The detected proteins are found only in females and eggs, and are essential for reproduction of this pest. This discovery is significant because it is the first report of detection of female-specific proteins by an antibody in a plant-parasitic nematode, provides the opportunity to study molecules that control reproduction in nematode pathogens of crop plants, and has potential for development into a specific nematode detection system for analysis of soil samples. This information will be used by researchers in the agrochemical and agricultural biotechnology industries who are developing safe and selective methods for nematode detection and control.
Technical Abstract: Antiserum was produced against the major female specific proteins, vitellogenins, of the soybean cyst nematode, Heterodera glycines. Denaturing polyacrylamide gel electrophoresis separations demonstrated two bands, corresponding to 180kDa and 190kDa estimated molecular weights under non-reducing sample conditions. Reduced samples yielded patterns of 190kDa and 205kDa apparent molecular weights, but no band at 180kDa. It is speculated that steric changes caused by reduction of sulfhydryl bonds in the proteins affected the electrophoretic mobility, and that two distinct vitellogenins are present. The antiserum detects vitellogenin in females and eggs but not in juveniles. The immunoblot pattern of eggs is more complex than in females, suggesting that some proteolytic processing may be involved in oocyte uptake of vitellogenin. Immunodetection by Western blot showed linear responses in both female and egg preparations, and was sensitive to as little as 10 egg equivalents. This suggests potential as a field survey or greenhouse detection tool.