Effector gene vap1based DGGE fingerprinting to assess variation within and among Globodera species and populations.

Abstract

Population-level genetic tools to study the epidemiology of potato cyst nematodes (PCN) are needed to evaluate their adaptive potential and validate management strategies. The effector gene vap1 is essential in early infection by PCN. We expected high allelic variation due to diversifying selection. A technique was developed specifically to amplify vap1 fragments of PCN, and electrophoretically separate variants in a denaturing gradient (DGGE). Sequenced gene variants differed in intron as well as exon regions. Siblings were more similar in vap1 pattern than juveniles from different cysts. Nearly all juveniles analysed had unique patterns with up to ten or five vap1 variants for Globodera pallida or G. rostochiensis, respectively. Patterns differed significantly among populations, with similarities increasing from between-species, within-species to within-pathotype comparisons. In conclusion, the method is suitable to determine the similarity of heterogeneous PCN populations for studies on their temporal change and spread.