An efficient method for identification of the Heterodera schachtii sensu stricto group using PCR with specific primers

S. Amiri, S. A. Subbotin, M. Moens

Abstract


A molecular method is described for the identification of nematode species from the Heterodera scbacblli sellsu stricto group. Based on the ITS-rONA sequence information obtained from more than 40 cyst forming nematode species, a primer SGRI was designed, which is specific for species from the H. schacbtii sensu stricto group. This specific primer in combination with the universal primer TW81 amplified a fragment of 850 bp from nematode DNA extracts. The primer was evaluated with 30 populations and species of the H. schachtii sellsu stricto group and several other parasitic nematode species. Subsequent digestion of amplified PCR products by Mval and Pvul allowed the separation of H. schachtii, H. betae and H. trifolii from each other. This method of identification is highly sensitive since amplification was obtained when a single second stage juvenile or <l single cyst were mixed with Pratylenchus penetrans or other nematode species.

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