Parasitology

Research Article

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

J. Roosiena1, P. M. Van Zandvoorta1, R. T. Folkertsmaa1, J. N. A. M. Rouppe Van Der Voorta1, A. Goversea1, F. J. Gommersa1 and J. Bakkera1

a1 Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands

Abstract

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.

(Received January 05 1993)

(Revised April 24 1993)

(Accepted April 24 1993)

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