General informationNotification NumberB/GB/09/R31/01Member State to which the notification was sentUnited KingdomDate of acknowledgement from the Member State Competent Authority04/01/2010Title of the ProjectControl of potato cyst-nematodes with minimised environmental impactProposed period of release:01/05/2010 to 30/11/2012Name of the Institute(s) or Company(ies)University of Leeds, Leeds LS2 9JT;
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:The organisms to be released are genetically modified potato plants that have been modified for improved resistance to Globodera pallida and Globodera rostochiensis, the two potato cyst nematodes that damage UK potato crops. Agrobacterium-mediated transformation of plants was used, which utilises the natural ability of the soil bacterium Agrobacterium tumefaciens to incorporate foreign DNA stably into the genotype of plants.
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The potatoes will express i) dsRNA with homology to digestive genes of two species of nematodes ii) dsRNA with homology to a potato gene involved in establishment and maintenance of the feeding cell iii) ribosome inactivating protein (RIP) from maize. None of the transgenes are lethal to potato cyst nematodes. The dsRNAs with homology to nematode genes limit ability of the nematodes to utilise dietary protein so slowing growth and suppressing egg production. The dsRNA with homology to the potato gene interferes with the nematode feeding cell and also slows growth and suppresses egg production. The RIP also interferes with the nematode feeding cell. Their expression will be under the control of CaMV35S from Cauliflower mosaic virus for constitutive expression or promoters that restrict expression. One root specific promoter to be used is from a serine threonine kinase (ARSK1) of Arabidopsis thaliana. A nematode-feeding site specific promoter from a Potato MIOX gene will also be used. Nos and ocs terminator sequences from Agrobacterium tumefaciens will terminate transcription of gene sequences. These sequences are not related to any technology that prevents seed propagation of plants.
All constructs will have the selectable marker gene for neomycin required only for selection of transgenic lines for evaluation and known to be biosafe as used.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:a. the selectable marker gene for resistance to neomycin from bacterial sources used to select transgenic lines after transformation
b. The frequently used promoter CaMV35S from Cauliflower mosaic virus for constitutive expression
c. the promoter from a serine threonine kinase (ARSK1) of Arabidopsis thaliana that provides differential expression of dsRNA in potato plant roots
d. a promoter from Potato that provides expression in the nematode feeding sites and anthers only
e. A region of the G. pallida cysteine proteinase gene expressed as an inverted repeat to form dsRNA in planta. Its effect suppresses growth and fecundity of potato cyst nematode without risk to humans in their diet or non-target organisms.
f. A region of the M. incognita cysteine proteinase gene expressed as an inverted repeat to form dsRNA in planta. It has no effect on growth and fecundity of potato cyst nematode and will be used as a control in the trial.
g. A region of the Potato MIOX gene expressed as an inverted repeat to form dsRNA in planta. Its effect suppresses growth and fecundity of potato cyst nematode without risk to humans in their diet or non-target organisms.
h. A b32RIP gene from Zea mays which suppresses growth and fecundity of potato cyst nematode without risk to humans in their diet or non-target organisms.
i. A nos terminator sequence from Agrobacterium tumefaciens that terminates transcription of gene sequences. This sequence is not related to any technology that prevents seed propagation of plants.
j. An ocs terminator sequence from Agrobacterium tumefaciens that terminates transcription of gene sequences. This sequence is not related to any technology that prevents seed propagation of plants.6. Brief description of the method used for the genetic modification:Standard procedures using Agrobacterium tumefaciens into which the constituent fragment had been cloned. Potato leaf tissue transformation leading to selection using antibiotic resistance and whole plant regeneration under contained conditions.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:This publicly funded research programme is not linked to commercial interests. The aims that this trial underpins are
a. To demonstrates the long term potential of biosafe novel resistance as a basis for control of potato cyst nematodes that replaces dependence on pesticide based control or the hidden costs of non-optimal cultural control
b. To demonstrate that such approaches are fully biosafe and environmentally benign
c. To demonstrate field efficacy for defences that have the ability to protect many crops from nematode pests
d. To demonstrate efficacy and biosafety before transfer to other projects in both Africa and Asia that aim to suppress nematode damage to subsistence crops. Nematodes cause an estimated untaken harvest in Africa alone that is sufficient to feed 100 million people.
The specific objectives of this proposed release are:
a. To test field performance of the novel potato plants against potato cyst-nematodes
b. To evaluate the efficacy of the resistance when delivered by specific promoters that limit expression primarily to where potato cyst nematodes feed in potato roots. The aim is to determine if resistance is as effective as when expression is throughout the plant. There are clear benefits to biosafety and the demand for new protein synthesis by the plant to restrict expression of even inherently safe proteins.
To determine that the transgenes have no adverse effects on free-living soil nematodes to add to the proven benign nature of cystatins that has already demonstrated2. Geographical location of the site:Headley Hall Farm, Nr Tadcaster, N. Yorkshire3. Size of the site (m2):1000 m24. 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:NoneEnvironmental Impact and Risk ManagementSummary of the potential environmental impact from the release of the GMPts:The possible selective advantage of cyst nematode resistance is of importance only in agricultural fields. It will not improve the survivability in the surrounding environment as potato cyst nematodes are restricted to fields with a history of growing potatoes. They are introduced pests with a limited host range. Four hundred years of cultivation of the potato has established that the potato has limited ability to survive in UK environments except when cultivated. Plants generated from tubers are readily eliminated and potato plants are not invasive of natural habitats. The pollen of potato normally disperses less than 10m and cannot cross with other crop plants to produce hybrids. The trial site will be considerably further than this isolation distance from other potato crops. The NPTII gene expressed in the potato plants imparts resistance to certain antibiotics of value during the selection process in tissue culture. This confers no selective advantage in the field and has been considered safe for such use by the European Food Safety Authority. It has a 13 year history of use with transgenic crops for this purpose. No difference with respect to persistence in agriculturally utilised habitats or invasiveness into natural habitats as compared to conventional potato varieties is expected. Through the measures which are taken during the release, distance from or absence of conventionally cultivated potatoes or wild species, the possibility of any gene transfer can be virtually ruled out. Even in the very improbable event that pollen was to be transferred to non-genetically modified potato plants, no consequences are to be expected, since potato propagation conventionally takes place via tubers and not via seeds. The modified potato lines will not have harmful interaction with non-target organisms. They are expected to have little impact even on other soil nematodes and it is an aim of the study to determine such impacts. No potato tubers from this trial will enter the food chain of humans or livestock and they will be destroyed at harvest. Measures which are taken under current release practice will both protect the trial against damage by wild animals, and also ensure that seed stock and plant material are harvested, stored, transported or disposed in such a way as to minimise or prevent contact with persons or animals. No effects on biogeochemical processes are expected, other than those which apply also to non-genetically modified potatoes.
There is an appreciable environmental benefit in the aim to provide an alternative to application of pre-plant pesticides to soil which is a common agricultural practice to control potato cyst nematodes and the largest variable cost of potato growing in the UK. The pesticides used are not specific for nematodes and they are harmful to other non-target organisms. There has also been a concern about groundwater contamination, pollution of aquatic systems with such chemicals and a consumer demand for no pesticide residues in water.Brief description of any measures taken for the management of risks:Conventional agricultural practices will be used at the field trial site. It will be at an isolation distance of more than 20 m from any field margin and considerably further from other potato crops. Pollination by bumblebees is highly local with any pollen collected from the transgenic plants deposited within the next few flowers visited. Therefore the transgenic plants will be surrounded by conventional potato plants to provide depositories for the pollen from the transgenic plants. The planting material will be brought directly from a containment facility in a van without other cargo. All tubers will be harvested by hand and be disposed of directly by a specialist, licensed company. . All herbage will be destroyed with a herbicide before harvest. Volunteers will be managed by monitoring their removal/destruction in land set to fallow and this will be maintained at monthly intervals until no volunteers have been detected for 3 consecutive months between April and October. In the unlikely event of unexpected plant growth once the trial has been planted, the whole area would be sprayed with an appropriate herbicide to destroy all plants and tubers lifted and destroyed by a specialist contractor.Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:Some of the plants to be used in the trial will express novel proteins under control of root specific promoters. This provides a proof of principle of nematode resistance without presence of the novel protein in ware tubers. The novel transgenes are inherently safe but no tubers from this work will enter either the human or livestock food chains. There is a clear environmental and food safety benefits in limiting expression of the novel expression to regions of the root system. There are also benefits to the plant in limiting the demand for new protein synthesis by the plant to roots.
The trial will develop bar coding of soil nematodes. Nematode communities have diverse trophic forms and so have the potential to be important bioindicators of soil health. This potential has not often been realised because of technical difficulties over rapid morphological identification. Bar coding overcomes this constraint. The approach provides a basis for detecting the benign or detrimental effect of any technology or agricultural practices on soil health.Final report-European Commission administrative informationConsent given by the Member State Competent Authority:Yes27/03/2010 00:00:00Remarks:Trial to take place at Headley Hall Farm near leeds, between 1 May and 30 November in 2010, 2011 and 20102. The area for the trial must not exceed 0.1ha each year.