Aphidiinae (Braconidae: Aphidiinae) is a subfamily of endoparasitic wasps specialized in parasitizing aphids. Although, to date, different methods have been used to measure parasitism level, obtaining an accurate estimate remains challenging due to several limiting factors. This study was set to: 1) Compare efficiency of conventional and molecular-based methods in estimating parasitism level of the pomegranate aphid Aphis punicae (Passerini; Hemiptera: Aphididae), and 2) Estimate seasonal activity of the Aphidiinae parasitoids of the pomegranate aphid. The molecular approach (polymerase chain reaction [PCR]) detected the presence of three main parasitoids Lysiphlebus fabarum (Marshal; Hymenoptra: Braconidae), Binodoxys angelicae (Haliday; Hymenoptra: Braconidae), and Ephedrus persicae (Frogatt; Hymenoptra: Braconidae). The presence of hyperparasitoid and aphid DNAs did not interfere with the outcome, indicating specificity of the selected primers. Minimum concentrations of DNA needed for successful amplifications were 16.33, 28.65, and 22.65 ng µl−1, for L. fabarum, B. angelicae, and E. persicae, respectively. The level of parasitism was significantly higher in spring (28.42%) than both summer and fall; parasitism level during summer (11.89%) and fall (5.86%) formed a homogeneous statistical subset. Although the overall level of parasitism estimated by PCR (22.7%) was more than twofold higher than those estimated by a conventional counting method (10.5%), there was a strong positive correlation between the two approaches. Provided the potential limitations of either method, simultaneous use of both methods was recommended for an objective estimate of the effectiveness of the Aphidiinae parasitoids as biological control agents of A. punicae.
Parasitoids play the positive role in aphids control management by keeping the population of aphids below the economic threshold. The pomegranate aphid, Aphis punicae is attacked by several parasitoids which mostly belong to Aphidiinae subfamily. The identification of parasitoids and hyperparasitoids is problematic due to their small size and ambiguous morphological characteristics. The analysis of molecular data of DNA sequences has been suggested as a complementary approach to the classical methods to identify biocontrol agents. In the present study, active parasitoids and hyperparasitoids of the pomegranate aphid collected from Razavi Khorasan province were identified using morphological characters and analysis of two gene sequences including COI and 28S. Phylogenetic analysis was performed using Maximum parsimony and Bayesian methods. The identified parasitoids were Binodoxys angelicae, Lysiphlebus fabarum, Ephedrus persicae, and the hyperparasitoids were Syrphophagus aphidivorus, Alloxysta sp. and Pachyneuron sp. We demonstrated that the mitochondrial COI gene was superior for the identification of these insects compared to the nuclear 28S (D2-region) and that it was compatible with morphological studies. This is the first study on molecular identification of the parasitoids and hyperparasitoids on pomegranate aphids in Iran.
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