DNA barcoding is a fast, accurate, and standardized method for species level identification, which is an important step in acarological and ecological studies. Traditional identifications often being based on the morphological characteristics. The morphological methods are not appropriate for identification of very small species, eggs, immature stages and cryptic species. Importance of oribatid mites in agriculture, lack of sufficient information on taxonomic status of these mites and controversial taxonomic position of some groups of this suborder, has recently gained a lot of interest to use molecular techniques beside morphological ones to help scientists in their researches. The genus Acrotritia was erected by Jacot (1923) based on the type species, Phthiracarus americanus Ewing, 1909. Members of this genus could be differentiated based on: the number of leg claws, number and shape of lateral carinae, shape of sensilli, shape and length of prodorsal and notogastral setae and also ornamentation of integument. Some of these characters can vary during slide mounting (shape of the sensilli) or lead to misidentification because they can be missed during slide preparation. Furthermore, some characters not stable in adult and immature stages (number of leg claws). So, establishment of a molecular technique could be helpful in this field. In this study, specimens of A. ardua were collected from soil of golden delicious apple orchards (Malus domestica; Rosaceae) and identified using morphological characters and then molecular techniques were applied to validate traditional identification. DNA was extracted from the preserved specimens by phenol-chloroform method. The D3 fragment of the 28S rDNA was partially amplified using the primer pairs D3A and D3B. The PCR cycling parameters consisted of a denaturation step at 95°C for 10 min followed by 35 alternating cycles of 30 second at 95°C for denaturation, 45 second at 54°C for annealing, 1 min at 72°C for extension. The fina
