Single juveniles of the potato cyst nematodes Globodera rostochiensis and G. pallida differentiated by randomly amplified polymorphic DNA.
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.
Roosien
J
J
Department of Nematology, Wageningen Agricultural University, The Netherlands.
Van Zandvoort
P M
PM
Folkertsma
R T
RT
Van der Voort
J N
JN
Goverse
A
A
Gommers
F J
FJ
Bakker
J
J
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers; D005260:Female; D008969:Molecular Sequence Data; D009348:Nematoda / Q000145:classification* / Q000235:genetics* / Q000254:growth & development; D016133:Polymerase Chain Reaction / Q000379:methods*; D011198:Solanum tuberosum; D013045:Species Specificity
D017931:DNA Primers; D004247:DNA