DNA sequences and cross-breeding experiments in the hawthorn spider mite Amphitetranychus viennensis reveal high genetic differentiation between Japanese and French populations

Sequence variation of the complete second internal transcribed spacer (ITS2, 445 bp) of nuclear ribosomal DNA and part of the mitochondrial cytochrome oxidase I gene (COI, 350 bp) was examined in Amphitetranychus viennensis (Zacher) mites (Acari:Tetranychidae) from four French and four Japanese locations. Sequence analysis consistently revealed the separation of the samples in two major groups: French mites differed from Japanese by 3.8-4.1% of the nucleotide divergence in COI sequences. These two groups also displayed distinct ITS2 consensus sequences (2.1% nucleotide divergence). A few variations, not affecting the diagnostic sites around the consensus sequence, were revealed among cloned copies of the same individual. Reciprocal crosses and backcrosses between one French and two Japanese populations disclosed strong reproductive incompatibility. However, fertile hybrid females were obtained, indicating the conspecificity of the tested mites. Despite the presence of Wolbachia in the French strain, but not in the Japanese ones, our crosses did not display the unidirectional incompatibility typically produced by this microorganism, but rather a bidirectional - although asymmetrical - incompatibility pattern. The post-zygotic incompatibilities in A. viennensis cannot be explained by the presence of Wolbachia but to some extent by mite genome divergence resulting from limited gene exchange between allopatric populations. Experiments of Wolbachia elimination by antibiotic treatment and subsequent crosses with cured strains are still needed to fully understand the reproductive incompatibility patterns in this mite species.