A rapid diagnostic technique of Bactrocera cucurbitae and Bactrocera zonata (Diptera: Tephritidae) for quarantine application

Abstract

Backround The melon fruit fly Bactrocera cucurbitae and the peach fruit fly Bactrocera zonata are serious pests, native to Asia, that have recently invaded Egypt. In Tunisia, no report of these species has yet been made, but pest risk data suggest that both Bactrocera species are likely to establish in other countries of the Mediterranean region. Results A quick method, based on a PCR‐RFLP of the mitochondrial COI gene, has been developed to enhance species identification for quarantine purposes. The restriction enzymes DdeI and XmnI were used to digest COI PCR products, enabling a rapid diagnosis of B. cucurbitae and B. zonata and their distinction from Tunisia's most devastating fruit pest, Ceratitis capitata. Conclusion The simplicity and relatively low cost of this molecular approach will replace the need to rear immature stages through to adults for identification and will facilitate rapid quarantine decisions providing greater plant protection.

Molecular Identification of Tephritid Fruit Flies (Diptera: Tephritidae) Infesting Sweet Oranges in Nsukka Agro-Ecological Zone, Nigeria, Based on PCR-RFLP of COI Gene and DNA Barcoding

“…Chua et al (2010) showed that this molecular method is effective even when only body parts or immature stages of tephritid fruit flies are present. Khemakhem et al (2012) successfully developed PCR-RFLP patterns of the COI gene in Tunisia to reliably identify and separate the quarantine pests B.curcubitae, B. zonata and C.capitata.…”

Molecular characterization of mariner-like elements in Bruchus pisorum and Bruchus rufimanus (Coleoptera: Bruchidae)

“…To authenticate the studied Bruchus species, we sequenced their DNA barcode region (COI). The COI mitochondrial DNA region was amplified for all samples using the universal primers designed by Folmer et al [24], LCO-1490: 5´TTTCTACAAATCATA-AAGATATTGG3´ and HCO-2198 5'TGATTTTTT-GGTCACCCTGAAGTTTA3´, and PCR conditions described by Mezghani et al [25]. The full-length MLEs specific to the mauritiana subfamily were amplified by the degenerate Mos1 primer 5'TAYCAGGRGTA-CAAGTAKGRAA3' described by Kharrat et al [26].…”

Loop-mediated isothermal amplification of economically important Ceratitis species (Diptera: Tephritidae)

“…In recent decades, molecular biology techniques have been continuously explored for the identification of fruit flies. These techniques include AFLP (amplified fragment length polymorphism) (Kakouli-Duarte et al, 2001), RFLP (restriction fragment length polymorphism) (Chua et al 2010, Mezghani Khemakhem et al 2013), RAPD (random amplified polymorphic DNA) (Sonvico et al 1996, Zhang et al 2004), DNA barcoding techniques (Zhang et al 2019, Li et al 2022), species-specific PCR and qPCR techniques (Jiang et al 2016, Koohkanzade et al 2018, Zheng et al 2019), etc. These tools are generally cumbersome, require rather expensive equipment and setup, and are generally time consuming.…”