The Aphidiinae subfamily (Hymenoptera: Braconidae) have significant role in biological control of aphids (Hemiptera: Aphididae). The successful use of biological control agents requires rapid and accurate identification of them. Identification and phylogenetic relationships studies of this subfamily have been performed based on morphological and behavioral information, but the new data is obtaining from molecular and embryological analysis. Nevertheless, phylogenetic relationships within this subfamily also did not resolve. For this propose alfalfa aphid parasitoids were coUected from different regions of North Khorasan province of Iran during 2012-2013. Identification of these wasps were performed using morphological characteristics and SEM images. Then, mitochondrial (COl and 16S) and nuclear (28S & LWRh) gene regions were amplified and sequenced. The resulted sequences were used for phylogeny reconstruction using two methods, MP and ML. In this survey, Aphidius ervi, Aphidius smithi, Aphidius matricariae, Praon barbatum, Praon volucre, Praon exsoletum, Lysiphlebus fabarum, Diaeretiella rapae and Trioxys complanatus species were identified. The results of molecular data showed that COl gene allowed more accurate identification of all the species rather than other genes. The phylogenetic analysis based on COl and 16S genes showed that this subfamily could be divided into four tribes (Aphidiini + Trioxini + Praini and Ephedrini), while they had tree tribes based on L WRh and 28S genes (Aphidiini (Aphidiina and Trioxina) + Praini and Ephedrini). The results of this study also showed the Ephedrini tribe as basal lineage based on Cor, 28S and LWRh, while 16S gene determined the Priani as basal tribe.

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