B: Diagnostics for high priority grains pests Dr Angela Freeman Senior Research Scientist Microbiology (DEPI)

Transcription

1 Project 2014: A: New tools for field grains surveillance Dr Jenny Davidson Senior Pulse Pathologist (SARDI) and B: Diagnostics for high priority grains pests Dr Angela Freeman Senior Research Scientist Microbiology (DEPI) Plant Biosecurity Cooperative Research Centre

2 Problem addressed- New diagnostic tools for high priority pests The DEPI- Victoria component of the project is focussing on the delivery of molecular diagnostics for high priority emergency plant pests (EPPs) of grains A major gap in diagnostic capability for exotic cereal viruses was identified Existing surveillance methods for identifying viral EPPs are time consuming and can be expensive both in terms of labour and the requirement for specialised laboratory facilities New and emerging viruses may be overlooked during border (PEQ) and postborder surveillance

3 Who will use the research and who will benefit directly from the research? The aim of the research is to reduce impacts of incursions of exotic pests on the grains industry by rapid detection and identification at the border and post-border At the border Partner with Department of Agriculture (DA) to develop improved post-entry quarantine protocols for cereals Direct benefits to the Australian Grains Genebank Plant breeders and seed companies developing new varieties Direct benefits to a range of diagnosticians, biosecurity agencies and researchers wishing to increase their ability to detect new and unknown pathogens in imported plant material using new molecular methods

4 Who will use the research and who will benefit directly from the research? The aim of the research is to reduce impacts of incursions of exotic pests on the grains industry by rapid detection and identification at the border and post-border Post-border Scientist, diagnosticians, agronomists and those involved in grains biosecurity and surveillance, breeding and disease management using molecular tests The broader surveillance network using rapid molecular tests and field based surveillance tools to enhance the detection and reporting of viruses from the field National biosecurity authorities (exotic incursion response, OCPPO identification of EPPs) State biosecurity authorities (regional outbreak of a restricted pest, control of interstate movements of material) The major beneficiary will be grain growers

5 Results so far review Conducted a review of rapid diagnostic capability for the top 35 high priority pests have identified gaps and a strategy to develop appropriate diagnostic tools for top 5 priority pests High Priority EPPs of grains will be incorporated into the Grains Industry Biosecurity Plan Identified gaps in diagnostic capabilities Identified a strategy to develop diagnostic tools for the top 5 high priority virus species (Polymyxa graminis as a vector) Preliminary discussions with federal Dept. of Agriculture on improved cereal quarantine

6 Results so far: New diagnostic tools for high priority pests New diagnostic tools Molecular diagnostic tests - A range of virus genus-specific and species-specific PCRs - Validation and publication of virus diagnostic protocols Surveillance (L. Zheng et al (2013) J. Virol Methods; submitted) - Survey of Victorian wheat crops in 2013 (2080 plant samples) - Screened for exotic and endemic cereal-infecting viruses using high-throughput molecular tools - Assessing a high-throughput, automated capillary electrophoresis gel system Map of survey sites Automated electrophoresis system Massive parallel sequencing (MPS) - MPS technology - simultaneous detection and identification of multiple pathogens (known and unknown) - Case study using clonal grass with a potential virus infection intercepted in post-entry quarantine Clonal grass showing viral symptoms intercepted at PEQ

7 Results so far: New diagnostic tools for high priority pests Field-based virus surveillance tools LAMP (Loop-mediated isothermal amplification) - A novel approach to nucleic acid amplification - Can be done in a small portable machine suitable for field surveys Progress - Evaluated published LAMP assays for a range of plant pathogens, including a bacterium(~30mins), a virus (~60 mins) and a viroid (~60mins) - Developed and evaluated a diagnostic assay for rapid detection of Barley yellow dwarf virus (BYDV) in the field at Horsham - LAMP was successful for the field detection of BYDV in wheat samples - Results presented at APPS conference in NZ (Zheng, L. et al (2013) 19th APPS Conference, Auckland NZ) 20 min 40 min

8 Challenges and issues arising from the research New diagnostic tools for priority EPPS - Diagnostics to support surveillance activities have traditionally been undertaken by experts in laboratories and/or in the field based on symptoms (e.g. grains) so the challenge will be to broaden the user base for field-based molecular diagnostics - The potential of new diagnostic platforms such as MPS and DNA fingerprinting are being assessed but the challenge will be in determining how these complex technologies fit into the Australian Biosecurity landscape

9 Prospects for success New diagnostic tools for high priority pests Technology success (molecular diagnostic tools and LAMP) Improved post-entry quarantine protocols for cereals Validated protocols for high priority EPPs (SPHDS) to ensure rapid response to incursions of EPPs

10 Thank you For more information, please A. B.