General informationNotification NumberB/DE/05/175Member State to which the notification was sentGermanyDate of acknowledgement from the Member State Competent Authority23/11/2005Title of the ProjectEvaluation of potato plants with increased tuber yield and starch content under field conditionsProposed period of release:04/04/2006 to 10/10/2007Name of the Institute(s) or Company(ies)University of Cologne, Albertus-Magnus-Platz, 50923 Cologne, Germany;
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:Modified potato plants appear normal in growth but have higher tuber yield and higher tuber starch content when grown under greenhouse conditions. This has been achieved by the introduction of two genes into the plant which code for metabolite translocators. The gpt (glucose-6-phosphate/phosphate translocator) gene of pea and the ntt1 (adenylate translocator 1) gene of Arabidopsis have been introduced and were controlled by the B33 patatin promoter which drives expression of genes mainly in potato tubers. The gpt gene has been introduced with the plasmid pBinB33(Hyg)::GPT. The selection marker here was the hpt gene from Streptomyces hygroscopicus conferring hygromycin resistance to transformed cells. The ntt1 gene has been introduced with the plasmid pBinB33(Kan)::NTT. The selection marker here was the nptII gene of Escherichia coli that confers kanamycin resistance to transformed cells. For 2 of 3 transformed plant lines evidence has been proven (using PCR technology) that the plants contain the nptIII gene, too. This gene is located outside the T-DNA of the plasmids and is driven by a bacterial promoter, i.e. although being present it is not expressed within the plants. The other above mentioned genes have been proven to be expressed by the existence of the activity of the respective gene products.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:T-DNA of pBinB33(Hyg)::GPT: patatin class I gene of potato (source) – promoter to drive expression of pea gpt gene (function); gpt gene of pea (source) – codes for a glucose-6-phosphate/phosphate translocator which transports glucose-6-phosphate in counter exchange with phosphate or triose phosphate across the inner envelope membrane of plastids (function); ocs gene of Agrobacterium tumefaciens (source) – transcription terminator for the gpt gene (function); nopalin synthase gene of Agrobacterium tumefaciens (source) – promoter to drive expression of hpt gene (function); hpt gene of Streptomyces hygroscopicus (source) – confers hygromycin resistance to transformed cells (function); gene 7 of Agrobacterium tumefaciens (source) – transcription terminator of the hpt gene (function).
|Common Name||Family Name||Genus||Species||Subspecies||Cultivar/breeding line|
T-DNA of pBinB33(Kan)::NTT: patatin class I gene of potato (source) – promoter to drive expression of Arabidopsis thaliana ntt1 gene (function); ntt1 gene of Arabidopsis thaliana (source) – codes for an adenylate translocator which transports ATP in counter exchange with ADP across the inner plastid envelope membrane (function); ocs gene of Agrobacterium tumefaciens (source) – transcription terminator for the ntt1 gene (function); nopalin synthase gene of Agrobacterium tumefaciens (source) – promoter to drive expression of nptII gene (function); nptII gene of Escherichia coli (source) – confers kanamycin resistance to transformed cells (function); nopalin synthase gene of Agrobacterium tumefaciens (source) – transcription terminator for the nptII gene (function).
Regions outside the T-DNA of pBin19-derived plasmids (pBinB33(Hyg)::GPT and pBinB33(Kan)::NTT are pBin19-derived plasmids): Since we proved evidence that an nptIII gene can be detected in 2 of 3 transformed lines, regions outside the T-DNA of both plasmids might be considered to be present in the plants. All the below mentioned genes or parts of genes should not be functional in plants: oriV of broad host range plasmid RK2 from Escherichia coli; part of the klaC gene of Klebsiella aerogenes; nptIII gene of Streptococcus faecalis; transposable element IS1 from Escherichia coli; trfA gene of broad host range plasmid RK2 from Escherichia coli; tetA gene (interrupted by the T-DNA) of broad host range plasmid RK2 from Escherichia coli; ColE1 ori of Escherichia coli; traF gene of broad host range plasmid RP4 from Escherichia coli; oriT of broad host range plasmid RP4 from Escherichia coli.6. Brief description of the method used for the genetic modification:Plants were transformed using Agrobacterium-mediated gene transfer: Leaves of sterile potato plants were cut and put into 25ml MS medium with 3.9% sucrose that contains a reduced 50ml over night culture of Agrobacterium tumefaciens GV2260 harbouring the plasmid pBinB33(Hyg)::GPT. Leaves were incubated for 48h in the dark. To induce calli, leaves were placed on MS medium containing 1.6% glucose, 0.5% naphtyl acetic acid, 0.01% benzamino purine, 0.05% cefotaxim and 0.3% gelrite in a plant incubator. After that leaves were transferred to MS medium containing 1.6% glucose, 2mg/l zeatin, 0.02% naphtyl acetic acid, 0.02 mg/l giberellic acid, 500mg/l cefotaxim, 3g/l gelrite and 100mg/l hygromycin. Incubation until shoots emerge, that can be cut and further cultivated to give a complete plant of which leaves can be taken to do a second round of transformation. This time agrobacteria contain the plasmid pBinB33(Kan)::NTT, and the antibiotic used for selection was kanamycin (50mg/l) instead of hygromycin. Lines used for further analyses did not display any growth of agrobacteria after several rounds of cutting and growing in tissue culture.7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:not applicableExperimental Release1. Purpose of the release:The purpose of the deliberate release of the transgenic potato plants into the environment in field trials solely is to verify the effects found in greenhouse experiments, i.e. increased tuber yield and starch content.2. Geographical location of the site:The release site is on the compound of the Max-Planck-Institute (MPIZ) in 50829 Cologne, district Cologne, federal state NRW, Germany. The site is not located in a flooded area, and is located in the local subdistrict Lövenich, field 54, cadastral parcel 00102.3. Size of the site (m2):1200 m2 (600 m2 per annum)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:not applicableEnvironmental Impact and Risk ManagementSummary of the potential environmental impact from the release of the GMPts:The genetically modified plants should not become more persistent in the agricultural habitat than the recipient plants, nor should they be more invasive in natural habitats. Transgenic potatoes differ from untransformed control plants only in tuber yield and starch content. Since there are potato cultivars that have a higher tuber starch content, and furthermore - dependent on the environmental conditions – tuber yield can differ strongly, there is no indication that these parameters determine persistence or invasiveness. In contrast, it has never been reported that potato plants can survive out of agricultural habitats in Europe. If transgenic plants are more persistent, we will be able to recognize this by monitoring the release site in the years after the release (see below). Under normal agricultural growth conditions, transgenic plants have no selective advantage or disadvantage compared to control plants, since they are not exposed to antibiotics. If a gene transfer to the same plant species occurs, this can only happen to the control plants on the release site, because other members of the species do not grow in the vicinity of the release site. These plants will be harvested and inactivated completely, and re-growth in the following years will be monitored and recorded. Sexually compatible plant species do not grow in Europe. The only Solanum species growing in Europe are Solanum dulcamara and Solanum nigrum, both can not be crossed with Solanum tuberosum or do not lead to viable hybrids. Effects on non-target organisms like pathogens can not be excluded. However, there`s no data available that tuber starch content influences plant pathogen interactions, and since starch content is not extraordinary high in the transgenic tubers, i.e. there are “naturally” existing cultivars with higher tuber starch content, there`s no reason to believe in a dramatic change in this kind of interaction. Gene transfer to fungi or bacteria may occur but is extremely unlikely. Micro-organisms must take up DNA / genes, and the uptake and incorporation into pre-existing plasmids or the genome is linked to a selective advantage for the organism. A theoretical advantage could be the uptake of an antibiotic resistance gene. An advantage can only occur if the gene taken up is expressed, and if there is selection pressure on the organism. Furthermore – a lot of micro-organisms already carry resistence genes, and it is much more likely that gene transfer from micro-organism to micro-organism occurs. A significant transgenic plant-born rise of resistent soil micro-organisms can be excluded.Brief description of any measures taken for the management of risks:The release site will be surrounded by a fence in order to keep small animals like rabbits and squirrels away from the transgenic plants. Bigger animals do not live on the compound of the MPIZ since this is surrounded by another high fence. A diversion of plant material can be avoided by this. Potato has a very low dispersal capacity and does not hybridise with any species growing wild in Europe. Thus, an outcrossing of the GM potato can be avoided by keeping a minimum distance of 20 m between the GM potato and any potato cultivation potentially growing on the agricultural area nearby. Monitoring by visual inspection during the plants are growing is warranted almost every day since workings to ensure good plant growth, e.g. chemical treatment against Phytophthora infestans or chemical and mechanical removal of weeds will be done. Unplanned incidents, e.g. death of plants will be recorded, the reason for such an incident will be discovered, if possible. Monitoring by visual inspection after growth of the plants will be done for at least two further years, so that potato plants emerging in these years can be recorded and killed. This is possible because during this time no potato plants will be grown at the site of release. All plants will be killed prior to harvest, and tubers will be digged out. Tubers will be either stored in appropriate facilities for the next year, will be used for analyses in appropriate laboratories or autoclaved to inactivate them.Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:not applicableFinal reportEuropean Commission administrative informationConsent given by the Member State Competent Authority:Yes11/05/2006 00:00:00Remarks: