Specificity of LAMP for genetic diagnostics of Chilean needle grass and serrated tussock

David Gopurenko, Aisuo Wang, Hanwen Wu

    Research output: Book chapter/Published conference paperConference paperpeer-review

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    Abstract

    Chilean needle grass (Nassella neesiana [Trin. & Rupr.] Barkworth) and serrated tussock (Nassella trichotoma [Nees] Hack. ex Arechav.) are related Weeds of National Significance (WoNS) undergoing rapid range expansion in Australia. They are difficult to distinguish from native tussock grasses and emergences of these weeds often remain undetected until after they have established as difficult to control populations. Genetic diagnostics using loop-mediated isothermal amplification (LAMP) may provide a simple cost-efficient means for rapid identification of these weeds, under laboratory and field conditions. Provision of LAMP diagnostics would assist in minimising the time-lag to initial recognition of tussock weed outbreaks in new areas, and subsequently improve the time to response for eradication of the weeds where they are recently emergent. Here we report laboratory development and tests of seven LAMP assays incorporating novel primer suites matched to species-specific DNA barcode sequences of Chilean needle grass and serrated tussock, and three additional suites designed for general LAMP validation. LAMP assays conducted using a real-time PCR platform tested positive for replicates of the target weeds at several of the primer suites and in less than20 min. Three of the suites are particularly promising(89–100% repeatability and specificity) and following optimisation to further minimise false positive or false negative test outcomes they are likely to be reliable for field use. As promising as these results are, the real challenge will be next-stage development of a simple but effective mobile LAMP platform for use in the field by officers engaged in weed surveillance efforts.
    Original languageEnglish
    Title of host publicationProceedings of the 21st Australian weed conference
    EditorsStephen Johnson, Leslie Weston, Hanwen Wu, Bruce Auld
    PublisherWeed Society of NSW
    Pages302-306
    Number of pages5
    ISBN (Print)9780995415911
    Publication statusPublished - 2018
    Event21st Australasian Weeds Conference (2018) - Novotel Sydney Manly Pacific, Sydney, Australia
    Duration: 09 Sep 201813 Sep 2018
    http://caws.org.nz/old-site/awc_contents.php?yr=2018 (conference proceedings)
    https://abercrombie.eventsair.com/QuickEventWebsitePortal/21awc/21awc-web/Agenda (conference program)

    Conference

    Conference21st Australasian Weeds Conference (2018)
    Abbreviated titleWeed biosecurity - Protecting our future
    Country/TerritoryAustralia
    CitySydney
    Period09/09/1813/09/18
    OtherThe Weed Society of New South Wales Inc., on behalf of the Council of Australasian Weed Societies Inc., will be hosting the 21st Australasian Weeds Conference in the popular Sydney beach side suburb of Manly from 9 - 12 September 2018.  An assortment of field trips will be hosted on Thursday 13 September 2018.
    This biennial conference carries on a long tradition of bringing the weed management community together to discuss new developments and share information about cutting-edge and best weed management practices.
    The conference attracts over 250 delegates from across Australasia and globally.  Delegates will come together to network with peers, engage with industry sponsors, listen and participate in presentations and field trips on a variety of topics including; 
    - New technologies in weed management.
    - Biological, mechanical, and chemical weed control and research.
    - Herbicide resistance.
    - Weeds of crops and pastures.
    - Environmental weeds and Weeds of National Significance.
    Internet address

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