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Single juveniles of the potato cyst nematodes Globodera rostochiensis and G. pallida differentiated by randomly amplified polymorphic DNA

Published online by Cambridge University Press:  06 April 2009

J. Roosien ,
P. M. Van Zandvoort ,
R. T. Folkertsma ,
J. N. A. M. Rouppe Van Der Voort ,
A. Goverse ,
F. J. Gommers  and
J. Bakker
J. Roosien
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
P. M. Van Zandvoort
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
R. T. Folkertsma
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
J. N. A. M. Rouppe Van Der Voort
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
A. Goverse
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
F. J. Gommers
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
J. Bakker
Affiliation:
Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands
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Summary

Random amplified polymorphic DNA (RAPD) offers a potential basis for the development of a diagnostic assay to differentiate the potato cyst nematode species Globodera rostochiensis and G. pallida. Nine decamer primers have been tested for their ability to amplify species-specific DNA sequences. Primer OPG-05 produced 2 discrete DNA fragments, which were consistently present in 5 G. rostochiensis populations and absent in 5 G. pallida populations. These fragments were detectable in single females as well as in single 2nd-stage juveniles. Their amplification is extremely efficient, and reproducible over a wide range of template concentrations. One-fifth of a single juvenile is sufficient to generate reproducible RAPD markers. The amplification from single juveniles requires no DNA isolation. The use of a crude homogenate does not impair the polymerase chain reaction.

Keywords

potato cyst nematodespathotypesrandom amplified polymorphic DNApolymerase chain reactionsibling species
Type
Research Article
Information
Parasitology , Volume 107 , Issue 5 , December 1993 , pp. 567 - 572
Copyright
Copyright © Cambridge University Press 1993

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