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Notification report


General information

Notification Number
B/GB/16/R8/01

Member State to which the notification was sent
United Kingdom

Date of acknowledgement from the Member State Competent Authority
26/01/2016

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

Proposed period of release:
01/04/2016 to 01/09/2017

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:
Two GM Camelina sativa lines 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.

These two different constructs are as follows. In the first iteration (B7.2), two genes from the picoalgae Ostreococcus tauri, and individual genes from the moss Physcomitrella patens, the Thraustochytriaceae Thraustochytrium sp., the oomycetes Phytophora infestans and Phytophora sojae and the diatom Thalassiosira pseudonana have each been linked to seed-specific regulatory sequences and introduced into the genome of Camelina sativa to direct the synthesis of EPA and DHA. In a second iteration (DHA2015.1), the sequence from Thalassiosira pseudonana was replaced with one from Ostreococcus RCC809, but in all other aspects was identical to B7.2. Both these constructs 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.

Both these constructs are very similar (differing only in one gene) with the Iteration C construct described in R8/14/01 and subsequently discussed in Usher et al., (2015) doi:10.1016/j.meteno.2015.04.002.


In parallel, a GM C. sativa line has been produced which accumulates the ketocarotenoid astaxanthin, through the introduction of the biosynthetic genes for this compound. Genes for the synthesis of astaxanthin are found in the higher plant Adonis aestivalis and two activities from this plant have been codon-optimized for expression in C. sativa, along with an additional gene from Zea mays. All three synthetic genes have been placed under the control of seed-specific promoters. This construct also contains the selectable marker bar which provides resistance to the inhibitor of glutamine synthase, bialaphos. The sequence of the bar gene is derived from Streptomyces hygroscopicus. This modification has not previously been described.


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 as detailed below.

Iteration B7.2.

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 thaliana 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
Tp4 1653bp Synthetic Encodes a fatty acid 4-desaturase from the marine diatom Thalassiosira pseudonana
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


Construct DHA2015.1


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 thaliana 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
O8094 1480bp Synthetic Encodes a fatty acid 4-desaturase from the marine picoalgae Ostreococcus RCC809
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








ASX-A2


Element Size Donor Organism Description and Intended Function

RB 24bp Agrobacterium tumefaciens T-DNA Right border
GLY 702bp Glycine max 11S Seed storage protein (Glycinin) promoter
HBFD1 1226bp Synthetic Encodes carotenoid 4-hydroxy-β-ring 4-dehydrogenase from Adonis aestivalis
Glyt 451bp Glycine max 11S storage protein (Glycinin) terminator
GLY 702bp Glycine max 11S Seed storage protein (Glycinin) promoter
ZmPhys 1232bp Synthetic Encodes phytoene synthase from maize
pNOS 288bp Agrobacterium tumefaciens Nopaline synthase gene promoter
bar 548bp Synthetic Encodes phosphinothricin acetyl transferase
NOSt 256bp Agrobacterium tumefaciens Nopaline synthase gene terminator sequence
GLY 702bp Glycine max 11S Seed storage protein (Glycinin) promoter
CBFD2 687bp Synthetic Encodes carotenoid β-ring 4-dehydrogenase from Adonis aestivalis
Glyt 451bp Glycine max 11S storage protein (Glycinin) terminator
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 on the basis of bialaphos 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:
We have been interested in developing an alternative, sustainable source of omega-3 long chain polyunsaturated fatty acids such as EPA and DHA (omega-3 fish oils) in transgenic plants, and have generated GM C. sativa plants which represent a terrestrial source of omega-3 long chain polyunsaturated fatty acids. Similarly, a line in which the ketocarotenoid astaxanthin is accumulated in the seeds has also been generated, and we have also produced a genetic cross between lines accumulating EPA and DHA, and lines accumulating astaxanthin. The purpose of this experimental trial is to determine the performance of different GM C. sativa iterations in the field, with respect to seed oil composition and oil quantity, and also to assess any additional phenotypic and agronomic variations. Specific questions to be examined are:

• Do the GM Camelina plants efficiently accumulate EPA and DHA in seed oil in the field?
• Do the GM Camelina plants efficiently accumulate astaxanthin in their seeds in the field?
• Do the crossed GM Camelina plants efficiently accumulate both EPA/DHA and astaxanthin in their seeds in the field?
• Do the GM Camelina plants still accumulate total seed oil to appropriate levels?
• Is there any further alteration to the lipidome of field-grown GM Camelina?
• Is there any difference between lines the different lines (including the genetic cross) in terms of agronomic performance?
• Is there any advantage or disadvantage to the GM Camelina 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 120130.

3. Size of the site (m2):
In Year 1 and Year 2, the total size of the trial site will be 2500m2.

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 four GM C. sativa lines are indistinguishable from the non-GM equivalent except for the modified composition of their seeds, in particular by the presence of the health-beneficial omega-3 long chain polyunsaturated fatty acids EPA and DHA and/or the ketocarotenoid astaxanthin. This modified 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.

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 area proposed to be planted with GMOs is small and temporary (lasting between 4 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 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.

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
18/04/2016 00:00:00
Remarks: