General informationNotification NumberB/GB/16/03Member State to which the notification was sentUnited KingdomDate of acknowledgement from the Member State Competent Authority11/03/2016Title of the ProjectInvestigating altered agronomic performance of wheatProposed period of release:03/01/2017 to 30/09/2018Name of the Institute(s) or Company(ies)Rothamsted Research, West Common, Harpenden
3. Is the same GMPt release planned elsewhere in the Community?NoHas the same GMPt been notified elsewhere by the same notifier?NoGenetically modified plantComplete name of the recipient or parental plant(s)
2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:Photosynthetic rate is one factor determining crop yield. This field trial investigates the effects of increasing the levels of a key enzyme in photosynthesis (SBPase) in wheat plants in the field. To achieve this, a gene sequence from a distantly-related grass species encoding the enzyme sedoheptulose-1,7-biphosphatase was added to these wheat plants. In addition, a selectable marker gene giving resistance to glufosinate herbicides is also expressed. There are no previous modifications.Genetic modification3. 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:Two plasmids were used.
|Common Name||Family Name||Genus||Species||Subspecies||Cultivar/breeding line|
Element Size Donor Organism Description and Intended Function
ColE1 724bp E.coli Origin of replication for plasmid replication in E. coli
pSa-Ori 484bp Agrobacterium tumefaciens Origin of replication for plasmid replication in Agrobacterium
nptI (aph(3’)-Ia) 812bp E.coli Bacterial selection gene conferring resistance to Kanamycin and other antibiotics
RB 25bp Agrobacterium tumefaciens T-DNA Right border
LB 24bp Agrobacterium tumefaciens T-DNA Left border
RTBVP 939bp Rice tungro bacilliform virus Promoter sequence from rice tungro bacilliform virus
SBPase 1367bp Brachypodium distachyon Coding sequence for expression of sedoheptulose-1,7-biphosphatase
CaMV35ST 233bp Cauliflower mosaic virus (CaMV) Terminator of the 35S viral transcript
Ubi+intron 1991bp Zea mays Maize ubiquitin 1 promoter + first intron driving constitutive expression in wheat
Bar coding sequence 581bp Streptomyces hygroscopicus Plant selectable marker gene encoding phosphinothricin acetyltransferase conferring resistance to herbicides with active ingredient glufosinate ammonium. (N.B. Bar gene inefficient/inoperative in this plasmid)
nos T 257bp Agrobacterium tumefaciens Nopaline synthase terminator
Element Size Donor Organism Description and Intended Function
ColE1 619bp E.coli Origin of replication for plasmid replication in E. coli
bla (beta-lactamase) 930bp E.coli Bacterial selection gene conferring resistance to ampicillin
Ubi+intron 1992bp Zea mays Maize ubiquitin 1 promoter + first intron driving constitutive expression in wheat
Bar coding sequence 541bp Streptomyces hygroscopicus Plant selectable marker gene encoding phosphinothricin acetyltransferase conferring resistance to herbicides with active ingredient glufosinate ammonium.
nos T 250bp Agrobacterium tumefaciens Nopaline synthase terminator for gene of interest6. Brief description of the method used for the genetic modification:Plasmid DNA was inserted into 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 Release1. Purpose of the release:The new gene added encodes an enzyme that functions to regenerate Ribulose-1,5-bisphosphate (RuBP) and the dephosphorylate sedoheptulose - 1,7-biphosphate (SBP) to sedoheptulose-7-phospate (S7P). Our hypothesis is that this will affect the rate of photosynthesis under field conditions.
We have studied these genetically modified plants in the laboratory and have already demonstrated that they possessed increased SBPase expression and showed several significant differences compared to controls including; enhanced maximum carboxylation efficiency of Rubisco (Vc,max) and regeneration of RuBP via photosynthetic electron transport (Jmax), increased vegetative biomass, and an increase in both number and total mass of seeds per plant (Raines / Parry, pers comm). This application seeks authority to investigate the effects of up-regulating the levels of SBPase in wheat plants in the field.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):The area for the proposed field trial, including controls and spacing between GM plots will cover 13.5m x 18m. A total area of approximately 240m2.4. 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 ManagementSummary of the potential environmental impact from the release of the GMPts:Observations on the general plant morphology of glasshouse-grown plants, timing of flowering, fertility, seed shape and germination show that the two GM wheat events Sox23 and Sox44 are indistinguishable from their non-GM equivalents except for the expected phenotype of enhanced SBPase expression and, under glasshouse growth conditions, increased total biomass and dry seed yield (unpublished data). No other changes to the plant morphology or development are apparent. The gene donor organisms (Brachypodium distachyon and Streptomyces hygroscopicus) are not known to be pathogenic or allergenic and neither the gene 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 20 metres from the boundary of the experimental plots 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 up-regulation of SBPase resulted in significantly enhanced photosynthesis, the chances of successful establishment of these wheat plants in unmanaged ecosystems is extremely low.
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 gene 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 1.8m x 6m plots (total area 86.4m2) and temporary (lasting between 5 and 6 months).
Although the above-ground plant material will be cleared from the site, the nptI 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 nptI 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 nptI 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 (Schluëter et al 1995, Nielsen et al 1997, EFSA, 2009), it cannot be completely discounted that some bacteria may successfully take up the nptI 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 nptI 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 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.
EFSA, 2009. Statement of EFSA on the consolidated presentation of the joint Scientific Opinion of the GMO and BIOHAZ Panels on the “Use of Antibiotic Resistance Genes as Marker Genes in Genetically Modified Plants” and the Scientific Opinion of the GMO Panel on “Consequences of the Opinion on the Use of Antibiotic Resistance Genes as Marker Genes in Genetically Modified Plants on Previous EFSA Assessments of Individual GM Plants”. The EFSA Journal 1108, 1-8. DOI: 10.2903/j.efsa.2009.1108 http://www.efsa.europa.eu/en/efsajournal/pub/1108
Nielsen, K.M., Gebhard, F., Smalla, K., Bones, A.M. and Van Elsas, J.D. (1997) Evaluation of possible horizontal gene transfer from transgenic plants to the soil bacterium Acinetobacter calcoaceticus BD413. Theor. Appl. Genet. 95, 815-821. DOI: 10.1007/s001220050630 http://link.springer.com/article/10.1007/s001220050630
Schluëter, K., Fuëtterer, J. and Potrykus, I. (1995) Horizontal gene transfer from a transgenic potato line to a bacterial pathogen (Erwinia chrysanthemi) occurs - if at all - at an extremely low frequency. Biotechnology 13, 94-98. PMID: 9636282Brief description of any measures taken for the management of risks:No wheat or other cereals will be grown within 20m 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 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 the 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 or removed by hand and destroyed.
Following completion of the 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 hand or the application of a systematic 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 informationConsent given by the Member State Competent Authority:Yes17/01/1930 00:00:00Remarks:To release wheat lines genetically modified for increased photosynthetic efficiency between 1 March 2017 and 31 December 2019.