Project Number: 1907-22000-018-12
Project Type: Reimbursable

Start Date: Jul 16, 2007
End Date: Sep 30, 2009

Objective:
1) Establishment of in vitro cultures for the potato cyst nematode (Globodera pallida) propagation. 2) Testing of molecular markers for G. pallida identification.

Approach:
-Both hairy root cultures of potato or tomato and regular root cultures of potato will be used to support the growth and the development of G. pallida. -Nematode will be surface-sterilized and inoculated on in vitro root cultures. -Once mature cysts are formed, they will be recovered and nematode reproduction rate and hatching ability will be evaluated. -Genomic DNA will be isolated from G. rostochiensis and G. pallida. -Primers specific to the chorismate mutase genes of G. rostochiensis and G. pallida will be used to amplify unique DNA fragments from both species. The amplified DNA fragments will be cloned and sequenced to confirm the specificity of these primer sets for nematode species identification. -Other nematode parasitism genes will be cloned from both nematode species and gene sequences will be analyzed to identify potential DNA polymorphisms that may be used for marker development.