Potato cyst nematodes from genus Globodera are common and widely distributed parasites of Solanaceae, mainly potatoes. Because of their persistence, large multiplication rate and low infestation levels, elimination of nematodes from field soils is difficult. Correct identification of the species of parasite can be hindered by high morphological resemblance of two main species infecting potatoes: G. pallida and G. rostochiensis. G. rostochiensis and
G. pallida, contrary to G. artemisiae, are considered by EPPO as quarantine pest, therefore distinction between these species is essential. We have undertaken attempts to differentiate between three Globodera species basing on the sequence of mitochondrial cytochrome oxidase (mtCOII) gene. It is also the first endeavor to analyse mtCOII genes of
G. rostochiensis and G. artemisiae; so far only one sequence of G. pallida COII gene is available in the GenBank database.

Total DNA of three Globodera species was isolated from intact cysts and used as a template in PCR reaction with specific primers, designed on the basis of the only available sequence of mtCOII from G. pallida. After condition adjustment, primers proved to amplify DNA from all three species. DNA fragments were isolated from agarose gel, cloned and sequenced. Obtained sequences were aligned and edited manually.

We observed numerous dissimilarities between species, confirming their evolutionary relationship. Sequences of G. pallida were in 96-97% similar to those deposited in GenBank. The observed similarity between species G. pallida and G. rostochiensis was 80-81%; between G. pallida and G. artemisiae – 70-71% and between G. rostochiensis and G. artemisiae – 71-73%.

The analysed COII fragment can be utilized for detection applications as well as for distinction between species. Analysis of mtCOII sequence can support or complement classic – laborious and time-consuming – methods for distinguishing between Globodera species.

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