Development of SNP-based assays for identification of Globodera rostochiensis and Globodera pallida

摘要

Potato cyst nematodes (PCNs), Globodera rostochiensis and G. pallida, are one of the major pathogens of potato and cause significant losses worldwide. These species, which are on the worldwide quarantine list, can survive in infected areas for more than 20 years. Therefore, these species need to be identified accurately and quickly. Although different molecular methods have been developed for the detection of G. rostochiensis and G. pallida, new techniques are being studied to identify these species. Kompetitive Allele-Specific PCR (KASP) based on single nucleotide polymorphisms (SNP) is a novel molecular technique. This technique is used for marker-assisted selection of genes that confer resistance in plants, for determining genetic variation of pathogens and for species identification. In this study, G. rostochiensis and G. pallida specific KASP primers were developed from the available DNA sequences of all Globodera species retrieved from GenBank. The KASP assay successfully identified the species G. rostochiensis and G. pallida. The developed KASP assays were able to distinguish these species from each other and from other nematode species. Results shown that even one cyst of these species can be sufficient for identification. This is the first study to develop KASP primers to identify PCNs. These assays can be used for accurate and rapid identification PCNs in laboratories.