Sequence analysis of the ribosomal Internal Transcribed Spacers region in spider mites (Prostigmata: Tetranychidae) occurring in citrus orchards in Eastern Spain: use for species discrimination

Tetranychus urticae is a polyphagous mite which is an important pest of citrus worldwide. This mite can be found feeding on many plant species occurring in the citrus agrosystem moving from weeds to trees. Because field samples consist of a mixture of different Tetranychidae species, as a first step necessary to further implement population characterization of T. urticae, species-discriminating criteria based on molecular techniques are needed. In this study, the nucleotide variation of the Internal Transcribed Spacers (ITS) 1 and 2 and the intergenic 5.8S fragment of nuclear ribosomal DNA of T. urticae, T. turkestani, T. evansi, T. ludeni and P. citri, have been determined. Results demostrate that for these species, the rDNA ITS2 regions are much more conserved than the corresponding rDNA ITS1. The high homogeneity of the ITS2 sequence observed among the specimens of T. urticae obtained from the same eco-region makes this DNA-sequience an excellent tool for species discrimination. ITS sequences differentiate not only species but also specimens from different geographical origin. Furthemore, PCR-RFLP analysis of the ITS2 proved adequate for a quick screening of high numbers of field sample