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DNA barcoding and real-time PCR detection of Bactrocera xanthodes (Tephritidae: Diptera) complex

  • D. Li (a1), D.W. Waite (a1) (a2), D.N. Gunawardana (a1), B. McCarthy (a3), D. Anderson (a3), A. Flynn (a1) and S. George (a1)...
Abstract

Immature fruit fly stages of the family Tephritidae are commonly intercepted on breadfruit from Pacific countries at the New Zealand border but are unable to be identified to the species level using morphological characters. Subsequent molecular identification showed that they belong to Bactrocera xanthodes, which is part of a species complex that includes Bactrocera paraxanthodes, Bactrocera neoxanthodes and an undescribed species. To establish a more reliable molecular identification system for B. xanthodes, a reference database of DNA barcode sequences for the 5’-fragment of COI gene region was constructed for B. xanthodes from Fiji, Samoa and Tonga. To better understand the species complex, B. neoxanthodes from Vanuatu and B. paraxanthodes from New Caledonia were also barcoded. Using the results of this analysis, real-time TaqMan polymerase chain reaction (PCR) assays for the detection of B. xanthodes complex and for the three individual species of the complex were developed and validated. The assay showed high specificity for the target species, with no cross-reaction observed for closely related organisms. Each of the real-time PCR assays is sensitive, detecting the target sequences at concentrations as low as ten copies µl−1 and can be used as either singleplex or multiplex formats. This real-time PCR assay for B. xanthodes has been successfully applied at the borders in New Zealand, leading to the rapid identification of intercepted Tephritidae eggs and larvae. The developed assays will be useful biosecurity tools for rapid detection of species in the B. xanthodes complex worldwide.

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*Author for correspondence Phone: +64 9 9098622 Fax: +64 9 909 5739 E-mail: dongmei.li@mpi.govt.nz
References
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Allwood, A., Tumukon, T., Tau, D. & Kassim, A. (1997) Fruit fly fauna in Vanuatu. pp. 7080 in Pacific, Allwood, A. & Drew, R.A.I. (Eds) Management of Fruit Flies in the a Regional Symposium, Nadi, Fiji. 28–31 October 1996. ACIAR Proceedings No. 76. Melbourne, Brown, Prior and Anderson.
Altschul, S. F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J. (1990) Basic local alignment search tool. Journal of Molecular Biology 215, 403410.
Armstrong, K. F. & Ball, S. L. (2005) DNA barcodes for biosecurity: invasive species identification. Philosophical Transactions of the Royal Society B: Biological Sciences 360, 18131823.
Armstrong, K. F., Cameron, C. M. & Frampton, E. R. (1997) Fruit fly (Diptera: Tephritidae) species identification: a rapid molecular diagnostic technique for quarantine application. Bulletin of Entomological Research 87, 111118.
Asokan, R., Rebijith, K. B., Singh, S. K., Sidhu, A. S., Siddharthan, S., Karanth, P. K., Ellango, R. & Ramamurthy, V. V. (2011) Molecular identification and phylogeny of Bactrocera species (Diptera: Tephritidae). Florida Entomologist 94, 10261035.
Blacket, M. J., Semeraro, L. & Malipatil, M. B. (2012) Barcoding Queensland Fruit Flies (Bactrocera tryoni): impediments and improvements. Molecular Ecology Resources 12, 428436. doi: 10.1111/j.1755-0998.2012.03124.x.
Bustin, S. A., Benes, V., Garson, J. A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M. W., Shipley, G. L., Vandesompele, J. & Wittwer, C. T. (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55, 611622.
Chua, T. H., Chong, Y. V. & Lim, S. H. (2010) Species determination of Malaysian Bactrocera pests using PCR-RFLP analyses (Diptera: Tephritidae). Pest Management Science 66, 379384. doi: 10.1002/ps.1886.
Cowley, J. M., Baker, R. T., Englberger, K. G. & Lang, T. G. (1991) Methyl Bromide Fumigation of Tongan Watermelons against Bactrocera xanthodes (Diptera: Tephritidae) and Analysis of Quarantine Security. Journal of Economic Entomology 84, 17631767.
Dhami, M. K., Gunawardana, D. N., Voice, D. & Kumarasinghe, L. (2016) A real-time PCR toolbox for accurate identification of invasive fruit fly species. Journal of Applied Entomology 140, 536552.
Drew, R. A. & Romig, M. C. (2001) The fruit fly fauna (Diptera: Tephritidae: Dacinae) of Bougainville, the Solomon Islands and Vanuatu. Austral Entomology 40,113150.
Drew, R. A. I. & Hancock, D. L. (1995) New Species, Subgenus and Records of Bactrocera Macquart from the South Pacific (Diptera: Tephritidae: Dacinae). Australian Journal of Entomology 34, 712.
Drew, R. A. I., AlIwood, A. J. & Tau, D. (1997) Bactrocera paraxanthodes Drew and Hancock – an example of how host records and attractant responses contribute to taxonomic research. pp. 131133 in Allwood, A. & Drew, R.A.I. (Eds) Management of Fruit Flies in the Pacific. A Regional Symposium, Nadi, Fiji. 28–31 October 1996. ACIAR Proceedings No. 76. Melbourne, Brown, Prior and Anderson.
Folmer, O., Black, M., Hoeh, W., Lutz, R. & Vrijenhoek, R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology Biotechnology 3, 294299.
Hebert, P. D. N., Ratnasingham, S. & deWaard, J. R. (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society B: Biological Sciences 270, S96S99.
Heimoana, V., Thnupopo, F., Toleafoa, E. & Fakanaiki, C. (1997) Fruit Fly Fauna of Tonga, Western Samoa, American, Samoa and Niue. pp. 5759 in Allwood, A. & Drew, R.A.I. (Eds) Management of Fruit Flies in the Pacific. A Regional Symposium, Nadi, Fiji. 28–31 October 1996. ACIAR Proceedings No. 76. Melbourne, Brown, Prior and Anderson.
Hoeben, P., Daniel, L. J., Ma, J. & Drew, R. A. I. (1996) The Bactrocera (Notodacus) xanthodes (Broun) species complex (Diptera: Tephritidae): comparison of 18S rRNA sequences from Fiji, Tonga and Vanuatu specimens suggest two distinct strains. Australian Journal of Entomology 35, 6164.
Jamnongluk, W., Baimai, V. & Kittayapong, P. (2003) Molecular phylogeny of tephritid fruit flies in the Bactrocera tau complex using the mitochondrial COI sequences. Genome 46, 112118. doi: 10.1139/g02-113.
Jiang, F., Li, Z. H., Deng, Y. L., Wu, J. J., Liu, R. S. & Buahom, N. (2013) Rapid diagnosis of the economically important fruit fly, Bactrocera correcta (Diptera: Tephritidae) based on a species-specific barcoding cytochrome oxidase I marker. Bulletin of Entomology Research 103, 363371. doi: 10.1017/S0007485312000806.
Jiang, F. Q., Liang, J. L., Zhang, A. B. & Li, Z. H. (2014) Existence of species complex largely reduced barcoding success for invasive species of Tephritidae: a case study in Bactrocera spp. Molecular Ecology Resources 14, 11141128.
Kunprom, C. & Pramual, P. (2016) DNA barcode variability and host plant usage of fruit flies (Diptera: Tephritidae) in Thailand. Genome 59, 792804. doi: 10.1139/gen-2015-0110.
Leblanc, L., Vueti, E. T., Drew, R. A. & Allwood, A. J. (2012) Host plant records for fruit flies (Diptera: Tephritidae: Dacini) in the Pacific Islands. Proceedings of the Hawaiian Entomological Society 44, 1153.
Leblanc, L., San Jose, M., Barr, N. & Rubinoff, D. (2015) A phylogenetic assessment of the polyphyletic nature and intraspecific color polymorphism in the Bactrocera dorsalis complex (Diptera, Tephritidae). Zookeys 26, 339367.
Li, D., Fan, Q.-H., Waite, D. W., Gunawardana, D., George, S. & Kumarasinghe, L. (2015) Development and Validation of a Real-Time PCR Assay for Rapid Detection of Two-Spotted Spider Mite, Tetranychus urticae (Acari: Tetranychidae). PLoS ONE 10(7), e0131887.
Meeyen, K., Sopaladawan, P. N. & Pramual, P. (2014) Population structure, population history and DNA barcoding of fruit fly Bactrocera latifrons (Hendel) (Diptera: Tephritidae). Entomological Science 17, 219230. doi: 10.1111/ens.12049.
OligoAnalyzer (2012) PrimerQuest® program. 3.1. Retrieved 12 December, 2012. Available online at http://www.idtdna.com/Scitools.
Plant Health Australia (2016) The Australian Handbook for the Identification of Fruit Flies version 2.1. 1/1 Phipps Close Deakin ACT 2600, Plant Health Australia.
R Development Core Team (2011) R: A Language and Environment for Statistical Computing. Available online at http://www.R-project.org/.
Stephenson, B.P., Gill, G.S.C., Ran, J.L. & Wilson, J. A. (2003) Biosecurity approaches to surveillance and response for new plant pest species. New Zealand Plant Protection 56, 59.
Tora Vueti, E., Allwood, A., Leweniqila, L., Ralulu, L., Balawakula, A., Malau, A., Sales, F. & Peleti, K. (1997 a) Fruit fly fauna in Fiji, Tuvalu, Wallis and Futuna, Tokelau and Nauru. pp. 6063 in Pacific, A. & Drew, R.A.I. (Eds) Management of Fruit Flies in the A Regional Symposium, Nadi, Fiji. 28–31 October 1996. ACIAR Proceedings No. 76. Melbourne, Brown, Prior and Anderson.
Tora Vueti, E., Ralulu, L., Walker, G., Allwood, A., Leweniqila, L. & Balawakula, A. (1997 b) Host availability – its impact on seasonal abundance of fruit flies. pp. 105110 in Pacific, A. & Drew, R.A.I. (Eds) Management of Fruit Flies in the A Regional Symposium, Nadi, Fiji. 28–31 October 1996. ACIAR Proceedings No. 76. Melbourne, Brown, Prior and Anderson.
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Bulletin of Entomological Research
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