Journal article
Robust environmental DNA assay development and validation: A case study with two vulnerable Australian fish
Aquatic conservation, Vol.32(7), pp.1225-1231
07/2022
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Abstract
Analysis of environmental (e)DNA can facilitate an understanding of the presence and distribution of aquatic species. However, eDNA detection using quantitative PCR requires validated and standardized species-specific assay designs. This study presents two eDNA assays to detect Murray cod, Maccullochella peelii, and mulloway, Argyrosomus japonicus (two ecologically vulnerable Australian species), based on small fragments of the mitochondrial 12S ribosomal RNA gene. A comprehensive description of species-specific assay development, from assay design to testing in silico, in vitro and in situ, has been included to guide effective assay design and validation in future studies. The results indicate that the assay was species specific for M. peelii within its natural distribution. However, the assay also amplified genomic DNA from two allopatric and endangered congeners (Maccullochella ikei and Maccullochella mariensis), thus potentially facilitating their eDNA detection elsewhere. In contrast, the A. japonicus assay was highly species specific with no amplification among close relatives. Both target-species assays are highly sensitive to as few as four and 10 copies per PCR reaction, respectively. This study has demonstrated that the assays assessed are effective tools for detecting the targeted species in situ from environmental DNA samples, which will assist efforts to conserve and manage their stocks.
Details
- Title
- Robust environmental DNA assay development and validation: A case study with two vulnerable Australian fish
- Creators
- Jackson Wilkes Walburn - New South Wales Department of Primary IndustriesMeaghan L. Rourke - New South Wales Department of Primary IndustriesElise Furlan - University of CanberraJoseph D. DiBattista - Australian MuseumMatt K. Broadhurst - New South Wales. Department of Primary IndustriesAshley M. Fowler - New South Wales Department of Primary IndustriesJulian M. Hughes - New South Wales Department of Primary IndustriesStewart Fielder - New South Wales Department of Primary Industries
- Publication Details
- Aquatic conservation, Vol.32(7), pp.1225-1231
- Publisher
- Wiley
- Number of pages
- 7
- Grant note
- 2019-016 / Fisheries Research and Development Corporation NSW Department of Primary Industries (DPI)
- Identifiers
- 991013093609902368
- Copyright
- © 2022 Commonwealth of Australia. Aquatic Conservation: Marine and Freshwater Ecosystems © 2022 John Wiley & Sons Ltd.
- Academic Unit
- Faculty of Science and Engineering
- Language
- English
- Resource Type
- Journal article