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On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
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DNA barcoding and real-time PCR detection of Bactrocera xanthodes (Tephritidae: Diptera) complex
D Li1, D W Waite1, D N Gunawardana1
1Plant Health and Environment Laboratory,Ministry for Primary Industries,P.O. Box 2095,Auckland 1140,New Zealand.
Bulletin of Entomological Research
|May 7, 2018
Summary
New molecular tools rapidly identify fruit flies at borders. Real-time PCR assays accurately detect Bactrocera xanthodes complex species, improving biosecurity for agricultural imports.
Area of Science:
- Entomology
- Molecular Biology
- Biosecurity
Background:
- Fruit fly larvae (Tephritidae) intercepted in New Zealand from Pacific nations are difficult to identify morphologically.
- Molecular identification revealed these belong to the Bactrocera xanthodes species complex, including related species like B. paraxanthodes and B. neoxanthodes.
Purpose of the Study:
- To develop a reliable molecular identification system for Bactrocera xanthodes and its related species.
- To create sensitive and specific real-time PCR assays for rapid detection of the B. xanthodes complex.
Main Methods:
- DNA barcoding of the COI gene for B. xanthodes, B. paraxanthodes, and B. neoxanthodes from various Pacific locations.
- Development and validation of real-time TaqMan PCR assays for singleplex and multiplex detection.
Main Results:
- A reference DNA barcode database was established for the B. xanthodes complex.
- Validated real-time PCR assays demonstrated high specificity and sensitivity (detecting down to 10 copies µl-1).
- No cross-reactivity was observed with closely related species.
Conclusions:
- Real-time PCR assays provide a rapid and accurate method for identifying species within the B. xanthodes complex.
- These assays have been successfully implemented at New Zealand borders for early detection of fruit fly eggs and larvae.
- The developed assays serve as valuable biosecurity tools for global pest management.
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