Abstract: Abstract：Objective   To verify whether there was tRNA gene loss in the mitochondrial genome of Trouessartia rubecula, on which process basis, a complete set of tRNA gene annotation process of mitochondrial genome in Astigmatina mites is proposed to provide a reference for the tRNA gene annotation of the mite mitochondrial genome. Methods   A series of tRNA annotation software, including MITOS, tRNAscan-SE, MITOS2 and ARWEN, were used in combination with manual sequence inspection, manual search,  and the minimum free energy (MFE) of possible secondary structures to verify the mitochondrial genome of Trouessartia rubecula. Results   The “deleted” genes of trnE, trnI and trnY in the mitochondrial genome of Trouessartia rubecula were successfully re-annotated based on the prediction results of MITOS and ARWEN combined with manual corrections. The trnA and trnV were found manually, and the re-annotated mitochondrial genome of Trouessartia rubecula indicated 22 tRNA genes. No loss of tRNA gene occurred. Conclusion   To obtain more accurate tRNA gene annotation results upon annotating mitochondrial genome in Astigmatina mites, tRNA gene prediction program should be widely used. And the prediction results of these programs should also be manually checked and corrected in order to reduce the bias caused by single program prediction and the annotation program prediction methods.

Key words: Trouessartia rubecula, Mitochondrial genome, tRNA genes annotation, Analgoidea, Astigmatina