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Notification report


General information

Notification Number
B/GB/14/R8/01

Member State to which the notification was sent
United Kingdom

Date of acknowledgement from the Member State Competent Authority
20/01/2014

Title of the Project
The synthesis of omega-3 long chain polyunsaturated fatty acids in Camelina sativa

Proposed period of release:
01/04/2014 to 30/09/2017

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
Gold-of-pleasure, false flaxBrassicaceaeCamelinasativaCeline

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
The GM Camelina sativa plants have been engineered with the novel capability to accumulate the non-native omega-3 long chain polyunsaturated fatty acids EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) through the introduction of the biosynthetic genes for these fatty acids. Such genes are normally only found in marine microbes such as microalgae and diatoms and some oomycetes and lower plants. Synthetic genes (meaning that the native DNA sequences have been codon-optimized and chemically synthesized) from EPA- & DHA-accumulating organisms have been integrated into the genome of Camelina sativa.

Three different constructs are described. In the first iteration (A), genes from the picoalgae Ostreococcus tauri, the moss Physcomitrella patens, the Thraustochytriaceae Thraustochytrium sp. and the oomycete Hyaloperonospora parasitica have each been linked to seed-specific regulatory sequences and introduced into the genome of Camelina sativa to direct the synthesis of EPA. In a second iteration (B), the first three sequences and regulatory elements are used in conjunction with single genes from both of the oomycetes, Phytophora infestans and Phytophora sojae, to direct the synthesis of EPA. In a third iteration (C), these same five genes present in B are used in conjunction with two additional genes, from Ostreococcus tauri and the coccolithophore Emiliana huxleyi. All seven of these biosynthetic genes are linked with appropriate plant-derived seed-specific regulatory elements, to direct the synthesis of EPA and DHA.

Two iterations (A, C) also contain the visual reporter protein DsRed, which allows for the simple identification of GM Camelina sativa seeds. The DsRed protein is derived from the marine coral species Discosoma sp and has been codon-optimised for expression in plants. One iteration (B) contains the selectable marker nptII which confers resistance to the antibiotic kanamycin, which was used to select GM Camelina plants. Kanamycin will not be used in the course of this field trial, and the presence of the nptII gene is not considered a risk in the context of this trial. 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:
Three plasmid constructs were used.

Iteration A.

Element Size Donor Organism Description and Intended Function

RB 24bp Agrobacterium tumefaciens T-DNA Right border

USP 684bp Vicia faba Unknown Seed Protein Seed-specific promoter

PSE1 873bp Synthetic Encodes an acyl-CoA-dependent 6-elongase from the moss Physcomitrella patens

35St 216bp Cauliflower mosaic virus 35S transcript terminator sequence

CNL 1064bp Linum usitatissimum 2S seed storage protein (Conlinin) promoter

Tc5 1320bp Synthetic Encodes a fatty acid 5-desaturase from the marine species Thraustochytrium

OCSt 192bp Agrobacterium tumefaciens octopine synthase gene terminator sequence

SBP 1800bp Arabidopsis Sucrose-binding protein promoter (seed-specific)

Ot6 1665bp Synthetic Encodes a fatty acid 6-desaturase from the marine picoalga Ostreococcus tauri

CatpAt 235bp Arabidopsis thaliana Cathepsin A gene terminator sequence

NP 664bp Brassica napus Napin seed specific promoter

Hpw3 1086bp Synthetic Encodes a fatty acid w3-desaturase from Hyaloperonospora parasitica

E9t 558bp Arabidopsis thaliana Ubiquitin E9 ligase gene terminator sequence

CsVMV 528bp Cassava vein mosaic virus Cassava vein mosaic virus (CsVMV) promoter

DsRed 684bp Synthetic Encodes a florescent protein from Discosoma spp.

NOSt 256bp Agrobacterium tumefaciens Nopaline synthase gene terminator sequence

LB 23bp Agrobacterium tumefaciens T-DNA Left border


Iteration B.

Element Size Donor Organism Description and Intended Function

RB 24bp Agrobacterium tumefaciens T-DNA Right border

USP 684bp Vicia faba Unknown Seed Protein Seed-specific promoter

PSE1 873bp Synthetic Encodes an acyl-CoA-dependent 6-elongase from the moss Physcomitrella patens

35St 216bp Cauliflower mosaic virus 35S transcript terminator sequence

CNL 1064bp Linum usitatissimum 2S seed storage protein (Conlinin) promoter

Tc5 1320bp Synthetic Encodes a fatty acid 5-desaturase from the marine species Thraustochytrium

OCSt 192bp Agrobacterium tumefaciens octopine synthase gene terminator sequence

SBP 1800bp Arabidopsis Sucrose-binding protein promoter (seed-specific)

Ot6 1665bp Synthetic Encodes a fatty acid 6-desaturase from the marine picoalga Ostreococcus tauri

CatpAt 235bp Arabidopsis thaliana Cathepsin A gene terminator sequence

NP 664bp Brassica napus Napin seed specific promoter

Piw3 1086bp Synthetic Encodes a fatty acid w3-desaturase from Phytophora infestans

E9t 558bp Arabidopsis thaliana Ubiquitin E9 ligase gene terminator sequence

NP 664bp Brassica napus Napin seed specific promoter

Ps12 1197bp Synthetic Encodes a fatty acid 12-desaturase FAD2 activity from Phytophora sojae

E9t 558bp Arabidopsis thaliana Ubiquitin E9 ligase gene terminator sequence

pNOS 288bp Agrobacterium tumefaciens Nopaline synthase gene promoter

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

NOSt 256bp Agrobacterium tumefaciens Nopaline synthase gene terminator sequence

LB 23bp Agrobacterium tumefaciens T-DNA Left border


Iteration C.

Element Size Donor Organism Description and Intended Function

RB 24bp Agrobacterium tumefaciens T-DNA Right border

USP 684bp Vicia faba Unknown Seed Protein Seed-specific promoter

PSE1 873bp Synthetic Encodes an acyl-CoA-dependent 6-elongase from the moss Physcomitrella patens

35St 216bp Cauliflower mosaic virus 35S transcript terminator sequence

CNL 1064bp Linum usitatissimum 2S seed storage protein (Conlinin) promoter

Tc5 1320bp Synthetic Encodes a fatty acid 5-desaturase from the marine species Thraustochytrium

OCSt 192bp Agrobacterium tumefaciens octopine synthase gene terminator sequence

SBP 1800bp Arabidopsis Sucrose-binding protein promoter (seed-specific)

Ot6 1665bp Synthetic Encodes a fatty acid 6-desaturase from the marine picoalga Ostreococcus tauri

CatpAt 235bp Arabidopsis thaliana Cathepsin A gene terminator sequence

NP 664bp Brassica napus Napin seed specific promoter

Piw3 1086bp Synthetic Encodes a fatty acid w3-desaturase from Phytophora infestans

E9t 558bp Arabidopsis thaliana Ubiquitin E9 ligase gene terminator sequence

NP 664bp Brassica napus Napin seed specific promoter

Ps12 1197bp Synthetic Encodes a fatty acid 12-desaturase FAD2 activity from Phytophora sojae

E9t 558bp Arabidopsis thaliana Ubiquitin E9 ligase gene terminator sequence

CNL 1064bp Linum usitatissimum 2S seed storage protein (Conlinin) promoter

OtElo5 903bp Synthetic Encodes an acyl-CoA dependent 5-elongase from the marine picoalgae Ostreococcus tauri

OCSt 192bp Agrobacterium tumefaciens octopine synthase gene terminator sequence

CNL 1064bp Linum usitatissimum 2S seed storage protein (Conlinin) promoter

Eh4 1380bp Synthetic Encodes a fatty acid 4-desaturase from the marine coccolithophore Emiliana huxleyi

OCSt 192bp Agrobacterium tumefaciens octopine synthase gene terminator sequence

CsVMV 528bp Cassava vein mosaic virus Cassava vein mosaic virus (CsVMV) promoter

DsRed 684bp Synthetic Encodes a florescent protein from Discosoma spp.

NOSt 256bp Agrobacterium tumefaciens Nopaline synthase gene terminator sequence

LB 23bp Agrobacterium tumefaciens T-DNA Left border


6. Brief description of the method used for the genetic modification:
Plasmid DNA was inserted into the C. sativa genome using Agrobacterium-mediated floral transformation system and transgenic plants were identified from the resulting seeds on the basis of kanamycin resistance or visual identification (DsRed expression)

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:
Purpose of the release (including any relevant information available at this stage) such as agronomic purposes, test of hybridisation, changed survivability or dissemination, test of effects on target or non-target organisms

Current sources of omega-3 long chain polyunsaturated fatty acids such as EPA and DHA are predominantly from oily fish, and it is for this reason these two fatty acids are often called fish oils. Global provision of fish oils is currently at or beyond the level of maximum sustainability, meaning that as the world’s population increases, there will be less of these health-beneficial fatty acids to go round. We have been interested in developing an alternative, sustainable source of fish oils in transgenic plants, and have produced GM C. Sativa plants which represent a terrestrial source of omega-3 long chain polyunsaturated fatty acids. The purpose of this experimental trial is to determine the performance of these three different GM C. Sativa iterations in the field, with respect to oil composition and oil quantity, and also to assess any additional phenotypic and agronomic variations. Specific questions to be examined are:

• Do the GM C. Sativa plants still accumulate fish oils in seed oil in the field?
• Do the GM C. Sativa plants still accumulate total seed oil to appropriate levels?
• Is there any further alteration to the lipidome of field-grown GM C. Sativa?
• Is there any difference between lines accumulating just EPA (iterations A, B) or EPA and DHA (iteration C) in terms of agronomic performance?
• Is there any advantage or disadvantage to the GM C. Sativa plants in terms of field-based performance?


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):
In Year 1, the total size of the site will be 900m2, doubling to 1800m2 in subsequent years. The layout of the plots will be duplicated in Years 2-4.

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 these C. sativa plants

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The three GM C. sativa lines are indistinguishable from the non-GM equivalent except for the modified fatty acid composition of their seeds, in particular by the presence of the health-beneficial omega-3 long chain polyunsaturated fatty acids EPA and DHA. This modified fatty acid composition is found only in the seeds of the GM C. sativa and is absent from all other vegetative tissues (e.g. leaves, roots, stems). The gene donor organisms are not known to be pathogenic or allergenic to humans, and none of the genes under investigation or the selectable or visual 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 occur since the C. sativa plants will not be consumed by humans.

The probability of C. sativa 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. C. sativa seeds are moderate in size and not normally dispersed by wind. Management measures including netting when the C. sativa is in flower 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 (such as insect-excluding netting) will further reduce the probability of these events occurring. There will be no compatible species grown for 1000 meters from the boundary of the site and no sexually-compatible wild relatives of C. sativa exist in the vicinity of the Rothamsted farm. In the unlikely event of a hybrid being generated, the presence of EPA and DHA in the seed oil of any such progeny will not convey a selectable advantage and most likely the omega-3 trait would not be retained.

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 marker genes would be expected to confer a selective advantage in the field environment under consideration. The genes introduced in C. sativa have been inserted via Agrobacterium tumefaciens-mediated gene transfer, and in one iteration (B) the insertion contains the bacterial nptII gene from E. coli, which is already widely present in the environment. The nptII gene expressed in the C. sativa plants imparts resistance to certain antibiotics, of value only during the selection process in culture. This confers no selective advantage in the field and has been considered safe for such use by the European Food Safety Authority and it has a 15 year history of use with transgenic crops for this purpose. 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 and temporary (lasting between 3 and 5 months).

Bearing in mind its limited scope, 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 C. sativa or related species will be grown within 1000m 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. Should the need arise to terminate the release at any point the emergency plans detailed below will be followed.

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 four-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.

In the unlikely event that the integrity of the site is seriously compromised, the trial will be terminated and all plants, (including GM and control C. sativa plots, and cereal separator) will be destroyed using a suitable herbicide or harvesting as deemed appropriate. All harvested material will be removed from the site and disposed of by incineration or deep burial at a local authority-approved landfill site using an approved contractor. Transportation of waste materials will be in secure containers. The phone numbers of all key staff will be available to site security and farm.


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
14/04/2014 00:00:00
Remarks:
This trial of Camelina sativa involves three independent transformation events, each of which has been genetically modified to express either four, five or seven genes originating from Thraustochytrium sp., Ostreococcus tauri, Physcomitrella patens, Phytophora infestans, Phytophora sojae, Emiliana huxleyi, or Hyaloperonospora parasitica. Together, these genes confer the ability to produce one or both of the omega-3 long-chain polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid in the seed oil of the plant. Two of the GMOs have also been modified to produce the selectable marker DsRed, the third has been modified to produce the antibiotic resistance marker NPTII.

The trial is authorised to take place at the Rothamsted Research farm near Harpenden, Herts. between 1st April 2014 and 31st October 2017.