The use of DNA microarrays for the developmental expression analysis of cDNAs from the oesophageal gland cell region of Heterodera glycines

Authors

  • Jan M. De Boer,

    1. Department of Plant Pathology, Iowa State University, 351 Bessey Hall, Ames, IA 50011, USA
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    • Present address: Laboratory of Plant Breeding, Wageningen University, PO Box 386, 6700 AD Wageningen, the Netherlands.

  • Jeff P. Mcdermott,

    1. Department of Plant Pathology, Iowa State University, 351 Bessey Hall, Ames, IA 50011, USA
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  • Xiaohong Wang,

    1. Department of Plant Pathology, North Carolina State University, Box 7616, Raleigh, NC 27695-7616, USA
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  • Tom Maier,

    1. Department of Plant Pathology, Iowa State University, 351 Bessey Hall, Ames, IA 50011, USA
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  • Fang Qui,

    1. Department of Agronomy, Iowa State University, Ames, IA 50011, USA
    2. Center for Plant Genomics, Iowa State University, Ames, IA 50011, USA
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  • Richard S. Hussey,

    1. Department of Plant Pathology, University of Georgia, Athens, GA 30602-7274, USA
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  • Eric L. Davis,

    1. Department of Plant Pathology, North Carolina State University, Box 7616, Raleigh, NC 27695-7616, USA
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  • Thomas J. Baum

    Corresponding author
    1. Department of Plant Pathology, Iowa State University, 351 Bessey Hall, Ames, IA 50011, USA
    2. Center for Plant Genomics, Iowa State University, Ames, IA 50011, USA
    3. Center for Plant Responses to Environmental Stress, Iowa State University, Ames, IA 50011, USA
      * Correspondence : Department of Plant Pathology, 351 Bessey Hall, Iowa State University, Ames, IA 50011, USA. E-mail: tbaum@iastate.edu
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* Correspondence : Department of Plant Pathology, 351 Bessey Hall, Iowa State University, Ames, IA 50011, USA. E-mail: tbaum@iastate.edu

Summary

A microarray was printed containing cDNAs from a library made from cytoplasm microaspirated from the oesophageal gland cell region of parasitic stages of the soybean cyst nematode, Heterodera glycines. The array contained both previously described clones (Wang et al. Mol. Plant-Microbe Interact. 2001, 14, 536–544) and uncharacterized cDNAs. Fluorescent probes for array hybridization were prepared using RNA polymerase amplification of nematode mRNA. Developmental expression profiles of the arrayed cDNAs were determined by hybridizing the microarray with probes from parasitic and non-parasitic H. glycines life stages. Distinct patterns of developmental expression were ascertained for the previously described gland expressed genes. In addition, four H. glycines cDNAs (SCN1018, SCN1020, SCN1028 and SCN1167) were identified that showed up-regulation in one or more parasitic life stages. Clone SCN1018 encodes a C-type lectin domain and is expressed in the hypodermis of females. Clone SCN1020 encodes a probable S-adenosylmethionine synthetase. Clone SCN1028 encodes a piwi protein with high similarity to the germ-line-specific protein R06C7.1 of Caenorhabditis elegans. The sequence of SCN1167 had no similarity to known genes. This paper describes the first use of cDNA microarrays to analyse genes of a plant-parasitic nematode and establishes a functional method to mine nematode cDNA libraries.

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