Tephritid fruit flies are ranked as one of the most damaging groups of insect pests. Morphological identification of fruit flies is mainly performed on adults due to the lack of adequate identification keys for immature stages. The peach fruit fly, Bactrocera zonata (Saunders), infests some of the principal commercial fruits and vegetables. It is, therefore important to avert its global dispersal, particularly by accurately identifying this species at ports of entry. In this study, a TaqMan real-time polymerase chain reaction (PCR) was developed for the accurate identification and sensitive detection of the peach fruit fly. A novel set of primers and probe were designed to specifically identify the mitochondrial cytochrome oxidase I (COI) gene. All specimens of peach fruit fly (including various life stages) were detected, and no cross reactivity with other tested tephritids were observed. Since this assay performed equally well with crushed insects and purified DNA, we note added efficiency by eliminating DNA extraction step. Considering the speed, specificity as well as sensitivity of the assay, Taqman real-time PCR can be used as a swift and specific method for pest species at ports of entry.

/pdf/rapid-identification-of-bactrocera-zonata-dip-tephritidae-4674th18do.pdf

Rapid identification of Bactrocera zonata (Dip.: Tephritidae) using TaqMan real-time PCR assay.

Of the seven species of Bactrocera fruit flies found in Samoa, only two (B. kirki (Froggatt) and B. xanthodes (Broun)) are of economic importance. These species attack a range of fruit, including papaya (Carica papaya), breadfruit (Artocarpus altilis), eggplants (Solanum melongena) and citrus. The presence of these two species limits export market access for Samoan produce. Eggplants and breadfruit infested with the eggs of B. kirki and B. xanthodes, respectively, were treated using a high-temperature forced-air (HTFA) protocol to heat the fruits to core temperatures of 40oC, 42oC, 44oC or 46oC. No B. xanthodes pupae emerged from fruit treated at 42oC or greater. Pupae of B. kirki were found from fruit treated at temperatures up to 44oC, but failed to survive treatments at 46oC. The HTFA protocol previously approved for treatment of other Pacific fruit flies (fruit core temperature to 47.2oC for 20 min) works without modification for treatment of the two combinations of fruit flies and commodities tested. However, less intense HTFA treatments are worth investigating, if required to enhance fruit quality.

/pdf/development-of-heat-treatments-for-two-species-of-samoan-4tjkjfgf70.pdf

Development of heat treatments for two species of Samoan fruit flies (Bactrocera spp., Diptera: Tephritidae).

ABSTRACTThrips (Order Thysanoptera) species are agriculturally important pests and vectors of plant disease. These tiny insects are commonly found in all life stages on imported commodities at the New Zealand border. Morphological identification of thrips is able to be performed on adults, but the identification keys for immature stages are inadequate and so DNA barcoding is regularly used for their identification. Here, we have generated cytochrome oxidase I (COI) DNA barcode data for 29 thrips species from 124 individuals, focusing on Frankliniella occidentalis, a dominant species intercepted at New Zealand border, followed by F. panamensis, Thrips palmi and T. tabaci. In addition, a multiplex real-time PCR assay has been developed to target the four thrips species. This assay is intended to facilitate the identification of quarantine interceptions with greater accuracy and faster diagnostic turnaround times, regardless of the developmental stages of the thrips. The developed assay demonstrated a high level of specificity for all the four target species and could detect as few as 10 copies/µL of the target DNA. Linear responses and high correlation coefficients between the amount of DNA and Cq values for each species were also achieved. The method was successfully tested on single egg, larva and adult samples and proved to be applicable for all life stages of the four species. Overall, this study has shown that the developed assay is an effective biosecurity tool for rapid and reliable identification of the target thrips species.KEYWORDS: Thripidaebiosecurityquarantine pestsborder detection AcknowledgementsThis work was part of the project (11 Int 03) funded by Operational Research programme from the Ministry for Primary Industries (MPI), New Zealand. We would like to thank all the Entomology staff at Plant Health and Environment Laboratory (PHEL), MPI for identifying the species, keeping the specimens, and performing the DNA extraction for this research. Our thanks go to Dr David Waite (PHEL, MPI) for conducting the internal review of the manuscript and his valuable suggestions. Great thanks also go to the editor and two anonymous reviewers for their constructive comments and edits to the manuscript, which has improved it significantly.Disclosure statementNo potential conflict of interest was reported by the author(s).Author contributionLK, DG, DL and AS conceived research.LK and DG secured funding.AS and DL designed the assay.AS, AT, YC and DL conducted all the experiments, including optimisation, specificity and sensitivity and blind panel testing of the assay.DG acquired thrips specimens and conducted morphological identification of the specimens.DL analysed the data and generated the figures.DL and AS wrote the draft manuscript. All authors read, edited and approved the manuscript.Data accessibility statementThe data used for this study were submitted into Zenodo, see xxx10.5281/zenodo.5748573 

DNA Barcodes for thrips species and development of multiplex real-time PCR assay for Frankliniella occidentalis Pergande, Frankliniella panamensis Hood, Thrips palmi Karny and Thrips tabaci Lindeman (Thysanoptera: Thripidae)

Zeugodacus cucumis and Bactrocera jarvisi are pests of fruit and vegetable crops and cause damage to horticulture industries. Immature stages of these two fruit fly species have been intercepted in New Zealand a number of times. Identification to species was not possible using morphological characters; thus, it is important to develop an assay for their species‐level identification. Here, the real‐time PCR assays for rapid identification of Z. cucumis and B. jarvisi were developed and validated. The PCR protocols demonstrated their specificity by amplifying the two target species successfully, with no cross‐reactions observed in the tested tephritid species. The in silico test of the primer and probe binding sites of the two assays also demonstrated the assays’ specificity by no mismatches present in the binding regions of the target species, but 1–4 mismatches in the binding regions of the non‐target fruit fly species. The thresholds of detection for the two assays are as low as 10 copies/µl of the target DNA, indicating that the assays have a very high sensitivity. The application of the real‐time PCR assays has greatly assisted in routine pest identifications at the New Zealand border and surveillance programme. Therefore, the assays have the potential to be used by diagnostic agencies and research organizations worldwide.

Real‐time PCR assays for rapid detection of Zeugodacus cucumis and Bactrocera jarvisi (Diptera: Tephritidae) for quarantine application

Fruit flies cause significant damage to crop and fruit production worldwide. Therefore, it is essential to identify these organisms to species level; however, immature stages are often impossible to be identified morphologically; thus, the application of DNA barcoding has greatly assisted in species identification. Nuclear, mitochondrial pseudo‐COI (NUMT) can be co‐amplified with mitochondrial DNA when using generic primers and therefore impair the efficacy of DNA barcoding. This study detected two types of NUMTs from Bactrocera passiflorae, one of them is novel. Therefore, the new finding will assist future species identification by avoiding misidentification using ambiguous NUMT sequences. In addition, this study has developed primers to target the COI gene of B. passiflorae, not the NUMT copies. The newly designed primers have demonstrated its efficiency in amplifying the Mt‐COI of B. passiflorae and can be used in routine diagnostics.

Molecular identification of Bactrocera passiflorae (Diptera: Tephritidae): Challenge and solution for DNA barcoding

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.

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

Handle everyday research tasks with reliable, citation-backed results

Your personal Research Agent to handle research tasks with citation-backed results

Popular Tasks used by Researchers

How can I help with your research?

Meet SciSpace

Get more enhanced response by uploading the PDFs you want me to reference.

No relevant PDFs in your library

SciSpace is the AI research assistant for academics. Run systematic literature reviews on 280M+ papers, and write papers with cited sources. Trusted by 1M+ students, PhDs & researchers.

SciSpace | AI for Research

Analyze PDFs

Code & Manuscripts

Funding & Grants

Literature & Patents

Medical & Clinical Data

Systematic Review

Visualize & Present

Web & Data

Build a Google Scholar-like website for your research.

Build a website

Create charts and images for your research

Create a Chart

Write a paper for submission to a journal

Draft a manuscript

Patent Search

Design eye-catching scientific posters in minutes.

Scientific Poster Generation

Systematic Literature Review

One task is running at the moment. Your messages will be shown right after.

Drag and drop or click here to browse

Loved by <highlight>1 million+</highlight> researchers

Extract a list of specific topics and their sources from unstructured text

Topics

Compare and analyze relevant papers that matches with your search

Papers

Get insights from PDFs and bookmarked papers from your library

My library

Recent searches

Try searching for:

Catch AI-generated content in scholarly and non-scholarly content

{ai} Detector

Ai Writer

Get PDF Summaries, highlighted text explanations 

Chat with PDF

Effortlessly create in-text citations and bibliographies in APA and 2,500 other formats

Citation generator

Get explanations, summaries, and answers on academic papers

Ease up your research workflow with {scispace}'s cohort of exciting AI tools

Elevate your academic writing skills and convey your ideas the way you want

Paraphraser

Explore our range of reading and writing tools

Your file is being prepared and should be ready in a few minutes. If it's a large file, it might take a bit longer. You can close this window, and we'll email you the file when it's done.

You have reached a maximum limit of {limit} columns in the table. Remove at least 1 column to add or create another one.

B McCarthy

Author Tools

Chat about Author

Papers

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