Session 6: On-farm DNA surveillance

biosecurity built on sciencePlant Biosecurity Cooperative Research Centre

PBCRC 2061

On-farm DNA surveillance for phylloxera – an end-user perspective

Inca PearceVinehealth Australia

Suzanne McLoughlinVinehealth Australia

Andy ClarkeYering Station

biosecurity built on science

A sensitive, accurate, cost-effective method for phylloxera detection for quantifying area freedom status

• “Where it is, where it is not”

Project aim

Department of Economic Development, Jobs, Transport and Resources

http://www.adelaide.edu.au/

http://www.depi.vic.gov.au/

http://www.depi.vic.gov.au/


Domestic313 million litres

$3.2 billion


Exports738 million litres119 countries$2.1 billion


Over 5,000 grape growers 135,000 hectares1.7 million tonnes


173,000 peopleregional areas

communities, suppliers, tourism


$40.2 billion gross output


The problem

biosecurity built on scienceThe impact


Well over half of the 135,000 hectares on own roots


Yering Station own vineyards – Yarra Valley


Maroondah Phylloxera Infested Zone

2006



2009 early



2009 late



2013



2015



2016



Current

biosecurity built on scienceImage courtesy of wine yarra valley


A draft method has been developed with the following key parameters

• Field sample timing:- Detection throughout the year → highest density in summer

• Field sampling for 1 hectare block:- Every 3rd row, every 5th panel- 40 cores

• Field sample collection:- Small dig stick- 5cm from vine trunk- 0-10cm depth of soil

• Field sample handling:- Composite- Cool- To lab for process to dry soil within 48 hours

• Laboratory analysis:- qPCR technique validated with sensitivity of 1-2 phylloxera per 200g soil

Following 3 years of research…


How does the DNA tool compare to other primary surveillance methods?

Preliminary survey of 24 vineyards identified 10 suitable on which to evaluate:• Sensitivity (proportion of positives correctly identified)• False positives • Cost • Time • Practicality

Comparative trial

DIGEMERGENCE

TRAPS DNA


Row 1

Row 4

Row 7

Row 10

Panel 2 Panel 7 Panel 13

31 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

1 23 4

3

3

3

3

3

3

3 1

2

5 vineyards only

4

3W1

2

W12 Dig

DNA

DNA double sampling rate

Emergence traps

W Weak vines


Performance of methods in detecting phylloxera

19 7 11 3

8 17 6 2

9 4

8 1

DNA and emergence trap equally as sensitive in detecting insects

and more sensitive than dig


Phylloxera DNA by qPCR

Laboratory

Soil/root sample with corer

Identify using microscope

Laboratory

Contents of trap in ethanol

Identify using hand lens

In field

An integrated approach to phylloxera detection and surveillance

Cost Scale Speed Sensitivity Accuracy Expertise required

DIG

Root sample with shovel

EMERGENCE TRAPS DNA

Sample collection

Sample processing

Method of phylloxera ID


Three primary methods for phylloxera detection

One method currently endorsed Use same sampling strategy in vineyard Important to overlay secondary methods of surveillance

DIGEMERGENCE

TRAPS DNA

Enhancing the potential of detecting phylloxera


Next steps for our researchProtocol development

(Nov ’16)

Securing diagnostic capability (May ‘17)

National endorsement (Apr ‘17)

Case study – surveillance at >block level (Mar’17)

Delivery model to end-users (May ‘17)

Extension to and adoption by end-users (Mar ‘18)

Integration into national and state protocols/regulations

(Sep ‘17)

1

2

3

45

6

7


REGULATORS• Early detection and ongoing surveillance• Establishment and maintenance of phylloxera zones• Enable better definition of areas of freedom (up and down)

GROWERS• Early detection, management and farm-gate hygiene• Potential for zoning change• Management of other pests & disease

INDUSTRY STAKEHOLDERS• Better defined zones will facilitate operations• Assist in quarantine process• ‘Clean, green, premium’ image maintained

RESEARCHERS• Provides a tool for studying the biology of phylloxera, rate of movement and vectors• Other soil borne pathogens and soil health

A supply chain view


For more information, please email [email protected]

Project team- Ray Correll (Rho Environmetrics)- Alan McKay (SARDI)- Kathy Ophel-Keller (SARDI)- Cassandra Collins (The University of Adelaide)- Danièle Giblot-Ducray (SARDI)- Greg King (DEDJTR-Vic)- Chris Anderson (DPI-NSW)- Inca Pearce (Vinehealth Australia)- Suzanne McLoughlin (Vinehealth Australia)

mailto:[email protected]

Session 6: On-farm DNA surveillance

Science

Transcript of Session 6: On-farm DNA surveillance