GM wheat at Rothamsted: Field trials and tribulations

Professor Johnathan Napier, Rothamsted

johnathan.napier@rothamsted.ac.uk

Barley yellow dwarf virus transmitted by cereal aphids

One of the cereal aphid species (Rhopalosiphum padi) on wheat

Aphids (greenfly / blackfly) are major pests on cereals and other crops: Feed on phloem sap and act as vector to spread viruses

Slide courtesy of Dr Toby Bruce RRes

Currently controlled by insecticides

Understanding how aphids communicate

Rothamsted has a long history of research into aphid communication via ‘smell’ (John Pickett et al.) – chemical ecology and plant-insect interactions

When aphids are attacked, either by predators or parasitiods, they produce an alarm pheromone which causes other aphids to move away Volatile sesquiterpene: (E)-Beta-farnesene

(EBF)

Lacewing larva (Chrysoperla carnea) attacking Aphis fabae Photos courtesy of Dr Toby Bruce RRes

Parasitic wasp: Aphidius ervi

Lacewing larva (Chrysoperla carnea) attacking (Myzus persicae)

7 Spot Ladybird (Coccinella septempunctata) (E)-Beta-farnesene (EBF) also ATTRACTS natural enemies of aphids

Chemical ecology operates on many (trophic) levels

repel pests

attract their enemies

Engineer plants to make EBF

Hypothesis:

Can we produce wheat plants that make EBF so they protect themselves?

Can we generate a new type of wheat that doesn’t need insecticides?

7

Project title:

A New Generation of Insect-resistant GM Crops:

Transgenic Wheat synthesising the Aphid Alarm Signal.

Applicants: John Pickett FRS Johnathan Napier Huw D Jones

£750,000

5 yrs.

2008 – 2013

(Biotechnology and Biological Sciences Research Council)

Funded by:

Investigators: Toby Bruce Gia Aradottir Lesley Smart Stephen Goward Angela Doherty

OPP

H

FPP (Farnesyl pyrophosphate)

EBF (E)-B-Farnesene

(E)-B-F synthase

FPP synthase

Biological and chemical ‘engineering’ to make wheat plants that produce EBF Generated 143 transgenic wheat plants (var. Cadenza) with 2 codon optimised (synthetic) genes that encode: EBF synthase (gene found in some plants (eg peppermint) FPP synthase (gene common in many plants & animals) (+ PAT enzyme as selectable marker)

PAT enzyme (herbicide tolerance) (Phosphinothricin acetyltransferase) Encoded by pat/bar genes

phosphinothricin (PPT) Gulfosinate ammonium Liberty Link, Basta etc

Inactivation via acetylation

Lab studies show these plants make EBF

1 6 . 0 0 1 8 . 0 0 2 0 . 0 0 2 2 . 0 0 2 4 . 0 0 2 6 . 0 0 2 8 . 0 0 3 0 . 0 0 3 2 . 0 0 3 4 . 0 0 3 6 . 0 0 3 8 . 0 0

5 0 0 0 0 0

1 0 0 0 0 0 0

1 5 0 0 0 0 0

2 0 0 0 0 0 0

2 5 0 0 0 0 0

3 0 0 0 0 0 0

3 5 0 0 0 0 0

4 0 0 0 0 0 0

4 5 0 0 0 0 0

T i m e - - >

A b u n d a n c e

T I C : 1 0 - 0 0 0 6 1 . D

(E)-Beta-farnesene

B2812 T2 EBF emission (ng/plant/hour)

0

500

1000

1500

2000

6 11 21 33 49 64 75 88

ng/

pla

nt/

ho

ur

Plant age (days)

Data courtesy of Dr Toby Bruce

Proceed to Field-scale evaluation of new aphid-repellent wheat via the emission of EBF Application for ‘part B’ (research-scale) release

• To UK ‘Department Food, Environment & Rural Affairs’ [Defra] - Advisory Committee on Releases to the Environment [ACRE]

• Comprehensive risk assessment focussing on human health and the environment (all online).

• Management plan. • Nothing to enter food/feed chain. All harvested material destroyed.

• 6-figure OS map reference to identify site.

• Included a 60 day public consultation period. • Wheat is a ‘good’ crop for GM trial because: No pollinating insects Self-pollinating (closed flower); pollen heavy /short viability No sexual compatibility with wild relatives Spring v winter types (temporal separation of flowering) Not an invasive or persistent species

GM Field Trial Application submitted to ACRE /Defra 20th June 2011

Legal notice in The Times 20th June 2011

Consent to carry out GM field trial received 16th Sept 2011

2012 GM wheat field trial design (not draw to scale)

4x4 Latin square 6m x 6m plots 2 GM lines plus control

6m 10 m separators (0.5m, 9m barley, 0.5m)

6m

1

GM1

2

Con

3

GM2

4

Con

5

GM2

6

Con

7

Con

8

GM1

9

Con

10

GM1

11

Con

12

GM2

13

Con

14

GM2

15

GM1

16

Con

80m

3m

6m wheat pollen barrier

80m

10m

10m

10 m separators (0.5m, 9m barley, 0.5m)

Trial sown on Rothamsted farm for 2 seasons: March - Sept 2012 March - Sept 2013 There will be two GM events tested. Total area = 6400 m2

Non-GM cereals = 6112 m2

GM area = 288 m2

The site had some security measures installed

Wheat (GM and control) was sown in the Spring

Representatives of the farming community were invited to visit the trial

We also consulted with local organic farmers and bee-keepers

So far, so good…..

The trial - mid summer: post-demo, experiment on-going

Logistically challenging !

Our field trial is large because it is an experiment

Year 1 – chemical ecology data successfully collected, trail harvested and site ready for Year 2 in 2013

36

Some personal thoughts on the first year of our GM wheat field trial • The media has become more sophisticated on the topic of GM • GM still causes a polarized debate, but perhaps not as much as before • The public are slightly “bored” with the topic • Social media is a powerful tool for public engagement

Specifics Our GM wheat trial had a number of interesting points • Rothamsted have no IP/patents for this trait • There is no industrial backer behind the project • There is the potential to reduce insecticide usage • Rothamsted has a strong record in the area of agro-ecology

Further information:

http://www.defra.gov.uk/news/ 2011/09/16/gm-wheat-trial/

http://www.rothamsted.ac.uk/Content.php?Section=AphidWheat

Email: field.trial@rothamsted.ac.uk Johnathan.napier@rothamsted.ac.uk John.pickett@rothamsted.ac.uk Huw.jones@rothamsted.ac.uk

38 http://www.temporarytemples.co.uk/imagelibrary/

Thank you!

Questions? Comments?

The next generation of insect repellent wheat?

The next challenge for us: Plenty more fish in the sea? The problem • Globally, fish accounts for the greatest proportion of protein that is consumed by humans. With such a high demand on this natural resource, many major fisheries – estimated to be greater than 70% - are being exploited at levels exceeding those that are sustainable. • Overfishing can reduce selected stock below levels that they can recover. As well as the catastrophic effects this can have on ecosystems and food-webs, it can severely affect livelihoods and economies. • Fish farming has been adopted as a method of meeting the demands of fish as a food source, but when poorly managed it can cause many problems including pollution, damage to the health of wild fish and creating ‘hotspots’ for disease.

http://www.zsl.org/conservation/news/plenty-more-fish-in-the-sea,594,NS.html

There is an urgent and compelling need for a sustainable source of fish oils • The majority of fish oils are now used by the aquaculture industry, which is a global production system expanding at ~10%/annum. • Aquaculture is well-recognised as having issues relating to sustainablity, and fish feed formulation companies are keen for alternative sources of fish oils • The value of fish oils is being driven up by the expansion of aquaculture, such that fish oils are now approximately 2X more valuable than vegetable oils.

We believe that transgenic plants

genetically engineered with the capacity to

synthesis omega-3 long chain PUFAs represent a solution to this problem