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Notification report


General information

Notification Number
B/GB/11/R8/01

Member State to which the notification was sent
United Kingdom

Date of acknowledgement from the Member State Competent Authority
20/06/2011

Title of the Project
Study of aphid, predator and parasitoid behaviour in wheat producing aphid alarm pheromone

Proposed period of release:
01/03/2012 to 30/09/2013

Name of the Institute(s) or Company(ies)
Rothamsted Research, West Common, Harpenden
Hertfordshire,
AL5 2JQ
UK;


3. Is the same GMPt release planned elsewhere in the Community?
No

Has the same GMPt been notified elsewhere by the same notifier?
No

Genetically modified plant

Complete name of the recipient or parental plant(s)
Common NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
wheatpoaceaetriticumtriticum aestivumCadenza

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
The introduced trait is increased resistance to aphids by expression of a natural alarm pheromone. The gene sequences that have been added to these wheat plants encode the enzymes (E)-β-farnesene synthase and farnesyl pyrophosphate synthase. In addition, the selectable marker genes (bar & nptI) giving resistance to glufosinate herbicides and the antibiotic kanamycin respectively are also present. There are no previous modifications.

Genetic modification

3. Type of genetic modification:
Insertion;

In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
Plasmid 1.
Element Size Donor Organism Description and Intended Function

ColE1 723bp E.coli Origin of replication for plasmid
replication in E.coli

nptI 800bp E.coli Bacterial selection gene conferring
aph(3’)-Ia) resistance to Kanamycin and other
antibiotics

RB 24bp Agrobacterium T-DNA Right border
tumefaciens

LB 23bp Agrobacterium T-DNA Left border
tumefaciens

Ubi+intron 2kb Zea mays Maize ubiquitin 1 promoter +
first intron driving constitutive
expression in wheat

EBFS 1.665kb Synthetic E-β farnesene synthase gene, codon
optimised for wheat

Chloroplast 147bp Wheat (Triticum Pre-sequence from the small subunit
targeting aestivum) of RubisCo to provide protein
sequence targeting to plastids (inc
chloroplasts)

nos T 300bp Agrobacterium Nopaline synthase terminator for
tumefaciens gene of interest

Bar coding 480bp Streptomyces Plant selectable marker gene encoding
sequence hygroscopicus Phosphinothricin acetyltransferase
conferring resistance to herbicides
with active ingredient glufosinate
ammonium. (N.B. Bar gene inefficient/
inoperative in this plasmid)

Plasmid 2.
Element Size Donor Organism Description and Intended Function

ColE1 723bp E.coli Origin of replication for plasmid
replication in E.coli

nptI 800bp E.coli Bacterial selection gene conferring
aph(3’)-Ia) resistance to Kanamycin and other
antibiotics

RB 24bp Agrobacterium T-DNA Right border
tumefaciens

LB 23bp Agrobacterium T-DNA Left border
tumefaciens

Ubi+intron 2kb Zea mays Maize ubiquitin 1 promoter +
first intron driving constitutive
expression in wheat

FPPS 1.074kb Synthetic A synthetic gene encoding farnesyl
pyrophosphate synthase, codon
optimised for wheat

Chloroplast 147bp Wheat (Triticum Pre-sequence from the small subunit
targeting aestivum) of RubisCo to provide protein
sequence targeting to plastids (inc
chloroplasts)

nos T 300bp Agrobacterium Nopaline synthase terminator for
tumefaciens gene of interest

Bar coding 480bp Streptomyces Plant selectable marker gene encoding
sequence hygroscopicus Phosphinothricin acetyltransferase
conferring resistance to herbicides
with active ingredient glufosinate
ammonium. (N.B. Bar gene inefficient/
inoperative in this plasmid)


6. Brief description of the method used for the genetic modification:
Plasmid DNA was inserted into wheat the wheat genome using a micro-particle delivery system and plants were regenerated using tissue culture methods.

7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:
Not applicable

Experimental Release

1. Purpose of the release:
The new genes added encode enzymes that lead to the production of a volatile chemical that is naturally produced by aphids and many other plants. This chemical, (E)-β–farnesene is known to repel aphids and to attract their natural enemies such as parasitoids and predators to the plant.

We have studied these genetically modified plants in the laboratory and have already demonstrated that they repel aphids and attract the natural parasitoids and predators. The purpose of this trial is to test whether these plants are better able to resist aphids under field conditions.

The aims of the trial are:

To evaluate the effect of the non-toxic aphid repellent (E)-β–farnesene as a approach to crop protection that does not rely on agricultural inputs (pesticides).
To confirm that (E)-β–farnesene that is produced by these GM wheat plants still functions in a ‘real life’ situation (i.e. in a field as opposed to a lab/greenhouse).
To determine the effect on colonization of the wheat crop by natural populations of three species of cereal aphid under real field conditions. We expect populations to be suppressed.
To evaluate effects on aphid specific predators and parasitoids (e.g. aphid parasitoids, ladybirds, hoverflies, lacewing larvae). These effects are expected to be positive unlike broad spectrum insecticides which do collateral damage to natural enemy populations
To determine effects on any other pest species present.
To investigate the persistence of plant DNA in the soil.


2. Geographical location of the site:
The release site will be located at Rothamsted Research, Harpenden, UK (OS grid reference TL 1213.

3. Size of the site (m2):
There will be eight 6x6m plots (288 m2) planted with GM wheat and including all the non-GM controls will cover a total area of approximately 6500m2

4. Relevant data regarding previous releases carried out with the same GM-plant, if any, specifically related to the potential environmental and human health impacts from the release:
There have been no previous releases of the same wheat plants

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The two GM lines are indistinguishable from their non-GM equivalents except for the volatile emission of (E)-ß-farnesene. Where applicable, the gene donor organisms are not known to be pathogenic or allergenic and neither the genes under investigation, nor the selectable marker genes are expected to result in the synthesis of products that are harmful to humans, other organisms or the environment. Any unknown hazards arising from the expression and ingestion of foreign proteins will not be realised because the wheat plants will not be consumed by humans.

The probability of seeds escaping from the trial site or the transfer of inserted characteristics to sexually-compatible species outside the trial area is estimated as very low. Commercial wheat varieties do not establish easily or thrive in uncultivated environments and are naturally self-pollinating with out-crossing being a rare event. Wheat seeds are relatively large and not normally dispersed by wind. Management measures including netting when the wheat is in ear and the use of gas guns and hawk kites will be employed to mitigate the risk of seed removal by birds. Management procedures to minimise the spread of seeds or pollen will further reduce the probability of these events occurring. There will be no cereals grown for 80 meters from the boundary of the site and no sexually-compatible wild relatives of wheat exist in the vicinity. If out-crossing to plants outside the trial area where to somehow occur, selection pressure to maintain the genes in the environment would exist only where glufosinate-based herbicides were applied. Even if the emission of (E)-ß-farnesene provides excellent protection from aphids at the field level (the trait under evaluation in these trials), the chances of successful establishment of these wheat plants in unmanaged ecosystems is extremely low and this would still be the case under severe infestations of aphids.
The risk of non-sexual, horizontal gene transfer to other species is extremely low. In the event of horizontal gene transfer to bacteria, neither the trait genes nor the selectable marker genes would be expected to confer a selective advantage in the field environment under consideration. The plasmid backbone sequences, nptI gene, origins of replication, border sequences etc. come originally from E coli and Agrobacterium tumefaciens, two common gut and soil bacteria respectively and these sequences are already widespread in the soil metagenome. Although this makes potential homologous recombination events more likely, we estimate the likelihood of horizontal gene transfer as low and the consequences, were it to occur, as negligible. The area proposed to be planted with GMOs is small; eight 6x6m plots and temporary (lasting between 5 and 6 months).
Although the above-ground plant material will be cleared from the site, the npt1 gene contained in the plant root DNA will decompose into the soil. The transgene is fully integrated into the plant DNA and the copy number is low thus the npt1 gene represents a very small proportion (much less than one millionth) of the total DNA in any one cell of our transformed wheat plants. This excess of competing DNA will significantly dilute the rate of any npt1 natural bacterial transformation. In addition, enzymatic degradation of free plant DNA in the soil and the low level of spontaneous bacterial competence to take up free DNA will significantly reduce the incidence of natural transformation. Although the transfer of functional gene units from plants to soil bacteria is accepted to be extremely low under natural conditions, it cannot be completely discounted that some bacteria may successfully take up the npt1 gene. However, there will be no antibiotics applied to the soil to provide additional selection pressure for the gene to persist in the environment. The source of the nptI gene is the gut bacterium E. coli carrying a plasmid containing the transposable element (Tn 903). R plasmids possessing resistance to aminoglycoside antibiotics are already naturally found in the soil and other environments. The npt1 gene encodes the enzyme Aminoglycoside 3’-phosphotransferase which confers resistance to kanamycin and related aminoglycoside antibiotics. Although these antibiotics still have some clinical applications, alternatives are readily available. Taken together, and bearing in mind the limited scope of this trial, the risk of generating of any additional antibiotic resistance within the soil microbial community or risks to human health or the environment if this were to occur as a result of the proposed trial is considered to be extremely low.
The overall risk of harm to human health or the environmental arising from this trial is assessed as very low.


Brief description of any measures taken for the management of risks:
No wheat or other cereals will be grown within 80m from the trial.

The release site will be visited by trained laboratory personnel who are working on the project at no less than weekly intervals (and at some periods, daily) during the growing season of each year of the trial. Any unexpected occurrences that could potentially result in adverse environmental effects or the possibility of adverse effects on human health will be notified to the Defra immediately.

In the unlikely event that the integrity of the site is seriously compromised or in other emergency situations, the trial will be terminated and all plants destroyed using a suitable herbicide or burning on site as deemed appropriate. Should the release site be subject to vandalism, care will be taken to ensure that all uprooted plant material within and outside of the trial site is identified and destroyed accordingly as described above.

At the end of each season, the plot will remain in stubble and monitored for volunteers during the remainder of the year and the following season. Any volunteers identified will be destroyed by herbicide treatment (e.g. glyphosate) or removed by hand and destroyed.

Following completion of the two-year trial the release site will remain fallow for a further season to enable easy identification of volunteers. The site will be inspected regularly and any volunteers identified will be immediately destroyed either by application of a systematic broad leaf herbicide.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Not applicable.

Final report
-

European Commission administrative information

Consent given by the Member State Competent Authority:
Yes
15/09/2011 00:00:00
Remarks:
Consent has been given for a trial of wheat designed to repel aphid pests by producing an aleam pheromone. The trial will take place at the Rothamsted Research site between March 2012 and September 2013, in 2 plots each not exceeding 6400 sq.m. A wheat pollen barrier of at least 2m width must surround the plots and during the release, cereals must not be grown in an area of at least 20m width surrounding the perimeter of the plots on which the GM wheat is planted. Measures must be taken to keep pigeons and other large birds out, and couch gass must be controlled.