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.

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.

Copyright Owner

The Society of Nematologists

Language

en

File Format

application/pdf

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