Campus Units
Plant Pathology and Microbiology
Document Type
Article
Publication Version
Published Version
Publication Date
9-1998
Journal or Book Title
Journal of Nematology
Volume
30
Issue
3
First Page
309
Last Page
312
Abstract
A method is presented for in-situ hybridization to mRNA in second-stage juveniles (J2) of the soybean cyst nematode Heterodera glycines. The protocol was developed using a digoxigenin-labeled RNA probe transcribed from cDNA of a cellulase gene that was known to be expressed in the subventral esophageal glands of H. glycines. Formaldehyde-fixed J2 were cut into sections with a vibrating razor blade to make the inside of the nematodes accessible for probing. These nematode fragments then were hybridized in suspension with riboprobe, and labeled with an alkaline phosphatase-conjugated antibody to digoxigenin. Staining with nitroblue tetrazolium and bromo-chloro-indolyl phosphate revealed a highly specific hybridization signal to mRNA within the cytoplasm of the subventral gland cells, using this specific antisense probe. This in-situ hybridization protocol will be useful for the characterization and identification of esophageal gland secretion genes in plant-parasitic nematodes, among other applications.
Copyright Owner
The Society of Nematologists
Copyright Date
1998
Language
en
File Format
application/pdf
Recommended Citation
de Boer, J. M.; Yan, Y.; Smant, G.; Davis, E. L.; and Baum, T. J., "In-situ Hybridization to Messenger RNA in Heterodera glycines" (1998). Plant Pathology and Microbiology Publications. 189.
https://lib.dr.iastate.edu/plantpath_pubs/189
Included in
Agricultural Science Commons, Agriculture Commons, Genetics and Genomics Commons, Plant Pathology Commons
Comments
This article is published as De Boer, J. M., Yitang Yan, G. Smant, E. L. Davis, and T. J. Baum. "In-situ hybridization to messenger RNA in Heterodera glycines." Journal of Nematology 30, no. 3 (1998): 309. Posted with permission.