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518 Florida Entomologist 92 ( 3 ) September 2009 MITOCHONDRIAL DNA MARKERS IN POPULATIONS OF DACUS PUNCTATIFRONS ( DIPTERA : TEPHRITIDAE )

Some of the true fruit flies (Diptera: Tephritidae) are major economic agricultural pests in several African countries (White & Elson-Harris 1992). Dacus punctatifrons (Karsch) (Subfamily Dacinae; Tribe Dacini) (Caroll et al. 2002) is a major pest of many cultivated and wild cucurbits as well as tomato ( Lycopersicum esculentum ). Even though, It is widely distributed in Sub-saharan Africa (Benin, Cameroon, Democratic Republic of Congo, and Equatorial Guinea) (Tindo & Tamo 1999), no information has been reported on its genetic make up. Surveys for Dacinae in Sub-Saharan Africa and identification of species including D. punctatifrons often were based on morphological and ecological characters (Mwatawala et al. 2006), but these methods have limitations, especially when the specimen is damaged or the adult stage is not available (Segura et al. 2006). Molecular taxonomy based on mitochondrial DNA has proved to be an efficient alternative to taxonomic identification (Muraji & Nakahara 2001). In fact, mitochondrial markers have been used with a number of insects for systematic and identification purposes (Barr & McPheron 2006; Segura et al. 2006; Virgilio et al. 2008).We have amplified and sequenced here a portion of the mitochondrial COII gene from various populations of D. punctatifrons . The specimens were collected at the adult stage and carefully identified basedon morphological features. Genomic DNA was extracted from individuals by the Lifton rapid fly genomic DNA isolation protocol as described in Anleitner & Haymer (1992). The polymerase chain reaction was used to amplify the mitochondrial COII sequences from each specimen with the following primers: