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A novel diagnostic gene region for distinguishing between two pest fruit flies:Bactrocera tryoni(Froggatt)and Bactrocera neohumeralis(Hardy)(Diptera:Tephritidae)

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Insect Science 2024年第4期31卷 1285-1295页

作者： Melissa L.Starkie Elizabeth V.Fowler Alexander M.Piper Xiaocheng Zhu Pauline Wyatt David Gopurenko Matt N.Krosch francesca strutt Karen F.Armstrong Hamish Patrick Mark K.Schutze Mark J.Blacket Department of Agriculture and Fisheries BiosecurityBrisbaneQueenslandAustralia Agriculture Victoria AgriBioBundooraVictoriaAustralia Department of Primary Industries Wagga Wagga Agricultural InstituteWagga WaggaNew South WalesAustralia Forensic Services Group Queensland Police ServiceBrisbaneQueenslandAustralia Department of Pest-management and Conservation Lincoln UniversityLincolnNew Zealand Better Border Biosecurity(B3 B3nz.org.nz)LincolnNew Zealand

Bactrocera tryoni and Bactrocera neohumeralis are morphologically similar sibling pest fruit fly species that possess different biological attributes,geographic distributions,and host *** need to differentiate between the two species is critical for accurate pest status assessment,management,biosecurity,and maintenance of reference *** morphologically similar,adults may be separated based on subtle characters;however,some characters exhibit intraspecific variability,creating overlap between the two ***,there is currently no single molecular marker or rapid diagnostic assay that can reliably distinguish between *** and ***;therefore,ambiguous samples remain *** we report the first molecular marker that can consistently distinguish between *** and *** diagnostic region consists of two adjacent single nucleotide polymorphisms(SNPs)within the pangolin(pan)gene *** confirmed the genotypes of each species are consistent across their distributional range,then developed a tetra-primer amplification refractory mutation system(ARMS)PCR assay for rapid diagnosis of the *** assay utilizes four primers in multiplex,with two outer universal primers,and two internal primers:one designed to target two adjacent SNPs(AA)present in *** and the other targeting adjacent SNPs present in ***(GG).The assay accurately discriminates between the two species,but their SNP genotypes are shared with other nontarget tephritid fruit fly ***,this assay is most suited to adult diagnostics where species confirmation is necessary in determining ambiguous surveillance trap catches;maintaining pure colony lines;and in Sterile Insect Technique management responses.

关键词： ARMS-PCR colony molecular diagnostics pangolin tetra-primer

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Development of internal COI primers to improve and extend barcoding of fruit flies (Diptera: Tephritidae: Dacini)

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Insect Science 2020年第1期27卷 143-158页

作者： Matt N.Krosch francesca strutt Mark J.Blacket Jana Batovska Melissa Starkie Anthony R.Clarke Stephen L.Cameron Mark K.Schutze School of Earth Environmental&Biological SciencesQueensland University of TechnologyBrisbaneQueenslandAustralia Plant Biosecurity Co-operative Research Centre BruceAustralian Capital TerritoryAustralia Agriculture Victoria AgriBioCentre for AgriBioscienceBundooraVictoriaAustralia Department of Entomology Purdue UniversityWest LafayetteIndianaUSA Department of Agriculture&Fisheries Queensland Primary Industries Insect Collection(QDPC)Dutton ParkQueenslandAustralia

Accurate species-level identifications underpin many aspects of basic and applied biology;however,identifications can be hampered by a lack of discriminating morphological characters,taxonomic expertise or *** approaches,such as DNA"barcoding"of the cytochrome c oxidase(COI)gene,are argued to overcome these ***,nuclear encoding of mitochondrial genes(numts)and poor amplification success of suboptimally preserved specimens can lead to erroneous *** insect group for which these molecular and morphological problems are significant are the dacine fruit flies(Diptera:Tephritidae:Dacini).We addressed these issues associated with COI barcoding in the dacines by first assessing several"universal"COI primers against public mitochondrial genome and numt sequences for dacine *** then modified a set of four primers that more closely matched true dacine COI sequence and amplified two overlapping portions of the COI barcode *** new primers were tested alongside universal primers on a selection of dacine species,including both fresh preserved and decades-old dry ***,Bactrocera tiyoni mitochondrial and nuclear genomes were compared to identify putative *** numt clades were identified,three of which were amplified using existing universal *** contrast,our new primers preferentially amplified the"true"mitochondrial COI barcode in all dacine species *** new primers also successfully amplified partial barcodes from dry specimens for which full length barcodes were *** we recommend these new primers be incorporated into the suites of primers used by diagnosticians and quarantine labs for the accurate identification of dacine species.

关键词： Bactrocera biosecurity Dacus\minibarcodes pseudogene Zeugodacus

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