Using point mutations in rDNA for differentiation of bioraces of Ditylenchus dipsaci from the Czech Republic
M. Zouhar, M. Marek, J. Lucinio, P. Ryšánekhttps://doi.org/10.17221/10490-PPSCitation:Zouhar M., Marek M., Lucinio J., Ryšánek P. (2002): Using point mutations in rDNA for differentiation of bioraces of Ditylenchus dipsaci from the Czech Republic. Plant Protect. Sci., 38: 358-360.
Ditylenchus dipsaci is one of the most harmful parasitic nematodes in Central Europe. It is able to survive for long time in soil without its host plants and that is why it belongs to organisms with quarantine importance. Nothing is known about D. dipsaci distribution in the Czech Republic. The aim of the study was to collect samples of D. dipsaci from the Czech Republic and to identify them by molecular methods. Region of rDNA including 3'end of 18 S gene, ITS1, 5,8 S gene, ITS2 and 5'end of 26 S gene was amplified using general primers designed according to the DNA sequence of Caenorhabditis elegans. The amplicon (900 bp) was analyzed by RFLP and SSCP. Restriction endonucleases Eco R1, Hinc II and Alu 1 can be used for differentiation of certain bioraces of D. dipsaci. At the same time methods for DNA extraction from plant material and contaminated soil were optimized.Keywords:
Ditylenchus dipsaci; DNA; SSCP; RFLP; PCR; biorace; differentiation