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Signal Peptide-Selection of cDNA Cloned Directly from the Esophageal Gland Cells of the Soybean Cyst Nematode Heterodera glycines
Molecular Plant-Microbe Interactions
  • Xiaohong Wang, North Carolina State University
  • Rex Allen, University of Georgia
  • Xiongfei Ding, University of Georgia
  • Melissa Goellner, North Carolina State University
  • Tom Maier, Iowa State University
  • Jan M. de Boer, Iowa State University
  • Thomas J. Baum, Iowa State University
  • Richard S. Hussey, University of Georgia
  • Eric L. Davis, North Carolina State University
Document Type
Article
Publication Version
Published Version
Publication Date
4-1-2001
DOI
10.1094/MPMI.2001.14.4.536
Abstract
Secretions from the esophageal gland cells of plantparasitic nematodes play critical roles in the nematodeparasitic cycle. A novel method to isolate cDNA encoding putative nematode secretory proteins was developed that utilizes mRNA for reverse transcription-polymerase chain reaction derived from microaspiration of the esophageal gland cell contents of parasitic stages of the soybean cyst nematode Heterodera glycines. The resulting H. glycines gland cell cDNA was cloned into the pRK18 vector, and plasmid DNA was transformed into a mutated yeast host for specific selection of cDNA inserts that encode proteins with functional signal peptides. Of the 223 cDNA clones recovered from selection in yeast, 97% of the clones encoded a predicted signal peptide. Fourteen unique cDNA clones hybridized to genomic DNA of H. glycines on Southern blots and, among them, nine cDNA clones encoded putative extracellular proteins, as predicted by PSORT II computer analysis. Four cDNA clones hybridized to transcripts within the dorsal esophageal gland cell of parasitic stages of H. glycines, and in situ hybridization within H. glycines was not detected for eight cDNA clones. The protocol provides a direct means to isolate potential plant-parasitic nematode esophageal gland secretory protein genes.
Comments

This article is published as Wang, Xiaohong, Rex Allen, Xiongfei Ding, Melissa Goellner, Tom Maier, Jan M. de Boer, Thomas J. Baum, Richard S. Hussey, and Eric L. Davis. "Signal peptide-selection of cDNA cloned directly from the esophageal gland cells of the soybean cyst nematode Heterodera glycines." Molecular Plant-Microbe Interactions 14, no. 4 (2001): 536-544, doi: 10.1094/MPMI.2001.14.4.536. Posted with permission.

Copyright Owner
The American Phytopathological Society
Language
en
File Format
application/pdf
Citation Information
Xiaohong Wang, Rex Allen, Xiongfei Ding, Melissa Goellner, et al.. "Signal Peptide-Selection of cDNA Cloned Directly from the Esophageal Gland Cells of the Soybean Cyst Nematode Heterodera glycines" Molecular Plant-Microbe Interactions Vol. 14 Iss. 4 (2001) p. 536 - 544
Available at: http://works.bepress.com/thomas-baum/50/