Member State to which the notification was sent
Date of acknowledgement from the Member State Competent Authority
Title of the Project
Genetically modified trees: consequences for biodiversity and ecosystem processes
Proposed period of release:
01/01/2008 to 31/12/2012
Name of the Institute(s) or Company(ies)
SLU Umeå, Umeå;
3. Is the same GMPt release planned elsewhere in the Community?
Has the same GMPt been notified elsewhere by the same notifier?
Genetically modified plant
Complete name of the recipient or parental plant(s)
INRA # 353-38 and INRA 717-1-B4
2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
INRA # 353-38 is modified to express the synthetic cry3Aa gene derivate from Bacillus thuringensis ssp. Tenebrionis cry3Aa gene. As a result the plants are resistant to insects especially Coleoptera. The toxin affects both growth and survival of the herbivorous beetles.The marker gene HPT from e-coli gives resistance to hygromycin INRA717-1-B4 has a downregulation of COMT and CAD. As a result the trees contain less lignin (COMT) or has changes in lignin structure (CAD) which improves pulping performance. The marker gene NPTII (neomycin fosfortransferas II gene) from e-coli gives resistance to neomycin
3. Type of genetic modification:
In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
INRA # 353-38 : a synthetic cry3Aa gene derivat from Bacillus thuringensis ssp. Tenebrionis cry3Aa gene was inserted. The intended function was to produce a toxin for insects (most specific to coleoptera). The origin of the terminator mas 3 is Agrobacterium tumefaciens, the HPT marker gene e-coli, the promotor CaMV Cauliflower mosaic virus; and the terminatorgene tml3 Agrobacterium tumefaciens. (see Fig 5) Fig. 5. Structure of binary vector pCGN4019. LB, left T-DNA border; RB, right T-DNA border; 35S, CaMV 35Spromoter; hygromycin B phosphotransferase (HPT) gene; tml, tml 3' region; Mac, hybrid 35S/mannopine synthetase promoter; cryllIA, rebuilt crylIIA gene; mas 3 ', mannopine synthetase 3' region.INRA 717-1-B4: is modified by that COMT cDNA were split into two fragments in a sense and antisense orientation and transferred with Agrobacterium tumefaciens. The intended function is to change the lignin concentration/structure and improve pulping performance. The origin of the terminatorgene 3’ocs is Agrobacterium tumefaciens, the nptII marker gene e-coli, the promotor CaMV Cauliflower mosaic virus; and PNOS-Agrobakterium (see fig below). We will only use one line (line ASCOMT2B) based on transformants from the plasmid pASB (antisense orientation) marked with * in the figure below
INRA717-1-B4 was also modified with CAD. The inserted CAD fragment has plant origin and the intended function is to reduce lignin content and change lignin structure to improve pulping performance. The origin of the terminatorgene 3’ocs is Agrobacterium tumefaciens, the nptII marker gene e-coli, the promotor CaMV Cauliflower mosaic virus; and PNOS-Agrobakterium (see fig below). We will only use one line (line ASCAD21) based on transformants from the plasmid p35SASCAD marked with * in the figure below.
6. Brief description of the method used for the genetic modification:
INRA # 353-38 is modified with Agrobacterium tumefaciens to express synthetic cry3Aa genderivat from Bacillus thuringensis ssp. Tenebrionis cry3Aa gene. The synthetic cry3Aa region flanked by the chimeric mac promoter (Comai et al. 1990), and the terminator mas3’ from pCGN4019 was provided by the Calgene company (Davis, CA) (Adang et al. 1993). It was excised as a XhoI fragment and inserted into the SalI site of the binary vector pBIBHygro (Becker 1990), resulting in the pBIBHCRY3Aa vector. This vector was introduced into the C58/pMP90 strain of Agrobacterium tumefaciens, by triparental mating. Poplar stem internodes were co-cultivated with the C58/ pMP90/pBIBHCRY3Aa strain of A. tumefaciens, as reported by Leplé et al. (1995). Transgenic calli were selected on a medium containing hygromycin (20 mg l−1).INRA717-1-B4: A poplar COMT cDNA was used to create various constructs under the control of the cauliflower mosaic virus (CaMV) 35S promoter (Leple et al. 1992). The COMT cDNA contained an intenal BamHI site, dividing the cDNA into two fragments (A and B) and both fragment were cloned into the binary vector pGSSJ780A behind the CAMV35s promoter in a sense and antisense orientation. Agrobacferium-mediated transformation (Leplé et al., 1992) was used to transform poplar (Populus tremula X Populus alba, clone Institut National de Ia Recherche Agronomique no. 717 1-B4). We will only use one line (line ASCOMT2B) based on transformants from the plasmid pASB (antisense orientation) (for more information see figure under paragraph B.4 above)INRA717-1-B4: The full-length CAD cDNA (cad.Pd X Pt.2) was isolated as a BamHI fragment. This fragment was inserted in both sense and antisense orientation into the pGSJ780A binary vector (harboring a T-DNA region containing a cauliflower mosaic virus 35s promoter-3’T7 and a pNos-NPTII-3’0CS chimeric gene) (Bowler et al., 1991) behind the cauliflower mosaic virus promoter. This resulted in two plasmids, p35SASCAD (the CAD fragment in antisense orientation) and p35SSCAD (the CAD fragment in sense orientation). Agrobacferium-mediated transformation (Leplé et al., 1992) was used to transform poplar (Populus tremula X Populus alba, clone Institut National de Ia Recherche Agronomique no. 717 1-B4). We will only use one line (line ASCAD21) based on transformants from the plasmid p35SASCAD (antisense orientation) (for more information see figure under paragraph B.4 above)
7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:
Populus spread by means of root suckers, wind dispersed pollen and seeds
1. Purpose of the release:
The aim is to test the effects of genetically modification on the biodiversity of associated organisms including both target and non-target organisms
2. Geographical location of the site:
SLU Campus in Umeå, Umeå Commune, Sweden
3. Size of the site (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:
The lignin modified line has been cultivated in France, from where no negative effects have been reported on the environment and on human health.
Environmental Impact and Risk Management
Summary of the potential environmental impact from the release of the GMPts:
If the Cry-gene would spread to native aspens it would be probably be a positive selective advantage for those individuals (as well as for planted Bt lines) as it would make them resistant to attacks by Coleoptera and maybe other insects. Other environmental impacts could be a reduced biodiversity on Bt aspens because of the toxcicity to herbivorous insects, such as leaf-feeding coleopteran. This could also influence higher trophic levels, such as predators (insects, spiders and birds) of e.g. coleopterans. Altered (reduced) ligning content would probably not be a selective advantage for the individuals trees (transgenic lines or native trees that acquired the genes by gene spread) as the lines contain less lignin, which probably reduces the stability of the trunk. However, the environmental benefit would include less use of chemicals and energy in the pulping process. The ecological impacts of GM aspens with reduced levels of lignin or changes in lignin structure is largely unknown but unintended effect of the transformation on plant traits (e.g. plant chemistry) could result in unforeseen effects on non-target species (e.g. Hjältén et al 2007). One of the objectives with this project is to investigate such effects. There is no risk that the plants used in this very short term experiment will have any negative environmental impacts or risk for human health (see E. for further information)
Brief description of any measures taken for the management of risks:
The plants are young, not more than 3 months – as a result they will not flower.They will be grown in pots on a plastic sheet, so that roots will not be in contact with the soilThe plants are too young and will be placed outside only during the summer months making it impossible for the roots to have grown to the size of being able to survive the winter, grow and re-sprout shoots de-attached from the mother plant. Closest wild relative is placed 100 m away. The ground will be checked – even the year after the last field trial
Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
The aim of this study is to study natural enemies in their natural environment; because it is only then we will be able to understand the real effects of the interactions. In this study we will be able to work simultaneously with several different types of enemies at the same time. The data will give us information on how the gm-aspens will affect target and non-target organisms.
European Commission administrative information
Consent given by the Member State Competent Authority: