Authors
Mark K. Nakhla,
Kristina J. Owens,
Wenbin Li, and
Gang Wei, National Plant Germplasm and Biotechnology Laboratory, United States Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) PPQ-CPHST;
Andrea M. Skantar, Nematology Laboratory, USDA Agricultural Research Service; and
Laurene Levy, National Plant Germplasm and Biotechnology Laboratory, USDA-APHIS-PPQ-CPHST, Beltsville, MD 20705
ABSTRACT
TaqMan primer-probe sets were developed for the detection and identification of potato cyst nematodes (PCNs) Globodera pallida and G. rostochiensis using two-tube, multiplex real-time polymerase chain reaction (PCR). One tube contained a primer-probe set specific for G. pallida (pale potato cyst nematode) multiplexed with another primer-probe set specific for G. rostochiensis (golden potato cyst nematode). A second tube consisted of the G. pallida-specific primer-probe set multiplexed with a primer-probe set specific for G. tabacum (the morphologically similar tobacco cyst nematode). This internal transcribed spacer rDNA-based system was specific for the Globodera spp. of interest and successfully identified several populations of PCN. This rapid, sensitive, and specific quantitative PCR assay presents a useful tool for PCN regulatory response and management programs.
