morphological and molecular identification of some indigenous fruit fly species in Sennar State, Sudan

Abdelaziz, E. Gesmallah, Massimiliano Virgilio, Marc De Meyer, Nabil H. H. Bashir, Mohamed E. Elkashif, Yousif O.H. Assad

Abstract


Fruit flies (Diptera:Tephritidae) are among the major constrains in commercial horticultural production in many African countries. DNA barcoding was used in this study to obtain quick and precise identification and to confirm the morphological identification of fruit fly specimens. Yellow sticky and Dome traps were used to collect some specimens from Sennar State whereas other specimens were reared under laboratory conditions at University of Gezira, from infested fruits. DNA from different specimens was extracted using the NucleoSpin® Tissue method. Three DNA fragments with sizes of 340, 220 and 280 bp. (base pairs) were recognized using specific primers and amplified from the 5’ region of the cox1 gene from the mitochondrial DNA to give a full barcode of 660 bp. Then, purified PCR products were subjected to sequencing reactions using the BigDye Cycle Sequencing Kit. The nucleotide sequences were aligned using the cluster W algorithm included in the Bioedit 7.0 software package. Sequence divergences were determined using similarity index (P-distance model). Morphologically, six fruit fly species, namely B. invadens, B. cucurbitae, C. cosyra, C. quinaria, D. ciliatus and D. longistylus were identified. The molecular phylo-genetic tree of the Sudanese fruit flies was established by sequencing of 12 specimens representing five fruit fly species at the mitochondrial cytochrome oxidase cox1 gene fragments. The morphological identification of the fruit fly

specimens was confirmed by molecular identification. Although B. invadens is a highly variable species, specimens of the same species

 

 

 

 

collected using yellow sticky traps, methyl eugenol and those reared

from magad fruits (wild plant belonging  to the family Cucurbitaceae) appeared to have the same or with a little variation (0.7%) in the DNA sequences cox1 fragments. D. ciliatus and B. cucurbitae appeared in separate clusters, with no variation among sequences of their specimens. The three common fruit fly genera: Bactrocera, Ceratitis and Dacus exhibited low intra-specific variation compared with that between inter-species in the amplified sequences.


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