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Notification report


General information

Notification Number
B/ES/11/26

Member State to which the notification was sent
Spain

Date of acknowledgement from the Member State Competent Authority
25/04/2011

Title of the Project
Production of transgenic trees with greater biomass. Release to the environment, field studies, environmental impact and risk assessment

Proposed period of release:
21/06/2011 to 21/09/2011

Name of the Institute(s) or Company(ies)
Universidad de Málaga (UMA), ;


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
hybrid aspensalicaceaepopuluspopulus alba x populus tremulaSelected clon from INRAVersailles (France) 7171-B4

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
It has been used as a vector for transformation pBI121 (Bevan, M (1984) Binary Agrobacterium vectors for plant transformation. Nucl. Ac. Res 12: 8711-8721), which is derived from a Ti plasmid of Agrobacterium tumefaciens which have deleted the genes involved in the synthesis of opines. Since the method and the vector used in transformation, the T-DNA region of plasmid pBI121 which contains the gene for the expression of Dof 5 from pine has been transferred into the genome of the hybrid P. X P.tremula alba.

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:
The chimeric gene contains the cDNA sequence of Dof 5 isolated from Pinus pinaster (Rueda-López et al. Plant Journal (2008) 56: 1973-1985) under the direction of 35S promoter of Cauliflower mosaic virus and linked to β-glucuronidase gene and a termination region of the nopaline synthase gene (NOS) from Agrobacterium tumefaciens (35S-Dof5-GUS-NOS). In addition to the Dof 5 chimeric gene, the plasmid also has a gene that confers resistance to kanamycin for selection of recombinant cells. This construct contains the sequence encoding neomycin phosphotransferase II (npt-II) from Escherichia coli flanked by the 35S promoter and the NOS region described above (35S-nptII gene-NOS). The size of both genes is 3.9 kbp (35S-GUS-NOS-Dof5) and 1.7 kbp (35S-nptII-NOS), respectively.

6. Brief description of the method used for the genetic modification:
Leaf segments were put in contact with the bacterial solution as described previously (Gallardo et al. Planta (1999) 210: 19-26 so the T-DNA region of plasmid pBI121 which contains the gene for the expression of Dof 5 pine has been transferred into the genome of the hybrid P. alba X P.tremula via Agrobacterium tumefaciens.

7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:
We have no information on the flow of transgenes from Populus to other organisms although this genre is being used in other countries for the generation of transgenic trees.

Experimental Release

1. Purpose of the release:
a) Perform an I + D project in collaboration with the company ENHOL SL, which will assess the field conditions the impact of genetic modification in the biology of trees. They are designed trials to understand the effects of over-expression of the transgene in the growth of trees and its relationship to seasonal variations in carbon and nitrogen content.
b) Evaluation of effects and potential risks that the cultivation of genetically modified trees could have on the environment. This evaluation is mandatory for any application for marketing of the transgenic product by the company ENHOL SL


2. Geographical location of the site:
Location: Southwest of Navarra (Spain), location : Buñuel.

3. Size of the site (m2):
6335 m2

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:
These plants have not been released into the environment before, however, plants grown in a similar method, using the same clone of INRA and which has carried out a field trial of three years was approved by the National Biosafety: Ministry of Environment (B/ES/98/27 reference) were not detected in any problems in the release (see article by Jing et al. New Phytologist, Special Issue: Populus Genomics 164, 137-145).

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
- Not previsible that the introduced traits can confer a greater selective advantage in natural environments.
- The PSMG has not acquired advantages or disadvantages.
- There is no information on the flow of transgenes from Populus to other organisms. Although this gender is being used in other countries for the generation of transgenic trees.
- Given the characteristics of the introduced genes and the possible use of trees in commercial activities (production of paper pulp and wood for various uses) is not expected potential danger or harmful effects on human health and on predators, parasitoids and pathogens in the environment.
- Not expected to have adverse interactions with the environment resulting from the expression of the transgene.
- Do not expect immediate or delayed effects on human health from direct or indirect interaction with PSMG.
- Not expected direct or indirect impact on the environment due to farming techniques, management and harvesting employed.
- Not expected effects on biogeochemical processes resulting from direct or indirect interactions.


Brief description of any measures taken for the management of risks:
1. Precautions taken:
a) distance or distances from sexually compatible plant species remain isolated experimental plot by a minimum safety distance of 10-15 meters in all respects.
b) measures to reduce or prevent the spread of pollen or seeds, the hybrid P. X P. tremula alba usually bloom between the fourth and fifth year of growth, are not covered by both measures to avoid seed dispersal. Trees were felled prior to the appearance of the flowers. To isolate the field experiment with other crops in the area will border the experimental plot with a mesh hutch.
2. Description of methods of treatment conducted after the release during the release will be deleted every 15 days herbaceous plants grown in experimental plots by mechanical removal and herbicide application in the planting rows. All material from the trees will be stored in bags for later incineration. The material collected will be held prior to irrigation of the plot due to flooding.
3. Description of the methods of post-release treatment in terms of waste collection of the GM plant.

The removal of the stems of the trees after harvesting, measurement and analysis for their study, they splinter and incinerated for total destruction. The removal of stumps after the second cut is made using a blender to a depth of 50 cm, which makes it impossible to regeneration with subsequent disposal of residual waste by incineration. The incineration of waste will be held at the installation of the dehydration fodder plant Enhol group have in the same place.

4. Description of the emergency plans.
As an emergency measure provides for the incineration of the plot. If such an eventuality the safety distance referred act as a firewall.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Description of monitoring plans and techniques.
The plants will be analysed to assess the stability of the introduced genes.
(1) evaluation for the presence of the transgene into the genome of poplar using two experimental approaches : PCR using specific primers, Also will be tested by Southern blot to confirm the presence of the transgene into the genome of the poplar and the number of copies inserted in each line.
(2) The transgene expression Dof5-35S-GUS-NOS using qPCR techniques to assess transcript levels in trees.
(3) The presence of the protein will be detected using monospecific antibodies generated in rabbits against the protein of pine using Western blot technique.
There will be a physiological and biochemical characterization of transformed lines during the early stages of tree growth. They are designed trials to understand the effects of over-expression of the transgene in tree growth, and its relationship to seasonal variations in carbon and nitrogen content.
Evaluation of the effects and potential risks that the cultivation of genetically modified trees could have on the environment.


Final report
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European Commission administrative information

Consent given by the Member State Competent Authority:
Yes
22/06/2012 00:00:00
Remarks:
The competent authority for give the consent of these field trials is the Autonomous Community of Navarra.