Back

Notification report


General information

Notification Number
B/DE/04/159

Member State to which the notification was sent
Germany

Date of acknowledgement from the Member State Competent Authority
20/09/2004

Title of the Project
Effect of stomata density on growth and yield of genetically modified potato in the agro-environment

Proposed period of release:
01/04/2005 to 30/11/2008

Name of the Institute(s) or Company(ies)
Max-Planck-Institute of Molecular Plant Physiology, ;


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
potatosolanaceaesolanumsolanum tuberosumtuberosumDésirée

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Potato plants transformed with the pBinARHyg-AtSDD1
- sequences from the subtilisin like serin protease SDD1 from Arabidopsis thaliana in sense orientation to the 35S promoter from Cauliflower mosaic virus
- OCS terminator from Agrobacterium tumefaciens
- Nos promoter from Agrobacterium tumefaciens
- hph gene from Streptomyces hygrocopius
- Nos teminator from Agrobacterium tumefaciens
- The transgenic plants may contain further parts of the vector pBinARHyg.
Plants transformed with the pBinAR-StSDD1
- sequences from the subtilisin like serin protease SDD1 gene of Solanum tuberosum in sense and antisense orientiation to the 35S promoter from Cauliflower mosaic virus, sense and antisense sequences are separated by an 54 bp sequence from the cloning vector pCR2.1_TOPO and thus form a sequence that acts as an RNAi-structure to the SDD1 gene when transcribed
- OCS terminator from Agrobacterium tumefaciens
- Nos promoter from Agrobacterium tumefaciens
- npt II (neomycin-phosphotransferase II) gene from Escherichia coli
- Nos teminator from Agrobacterium tumefaciens
- The transgenic plant may contain further parts of the vector pBinAR.


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:
Two groups of potato plants are intended for release.
Group 1 was transformed with the pBinARHyg-AtSDD1 construct, in which the subtilisin like serin protease SDD1 from Arabidopsis thaliana is in sense orientation to the promoter of the 35S-promotor of the cauliflower mosaic virus, terminated by the OCS terminator from Agrobacterium tumefaciens. The expression of the SDD1 gene reduces the number of stomata in the leaves of the plants. Under climate chamber condititions, the transgenic plants showed an increased sensitivity to high light intensities. The transformed potato plants further contain the hph gene from Streptomyces hygrocopius under the control of the Nos promotor and terminator from Agrobacterium tumesfaciens. The hph gene was introduced as a selection marker to facilitate the isolation of transgenic plants during the transformation process. Plant cells expressing the hph gene show an increased resistance to the antibiotic Hygromycin as compared to plant cells without hph expression.

Group 2 was transformed with the pBinAR-StSDD1 construct, in which the Subtilisin like serin protease SDD1 from Solanum tuberosum is in sense and partial antisense RNAi orientation to the promoter of the 35S-RNA of the cauliflower mosaic virus, terminated by the OCS terminator from Agrobacterium tumefaciens. The expression of the RNAi construct of the SDD1 gene increases the number of stomata in the leaves of the plants. Under high light or high temperature conditions, these plants showed an increased tuber production compared to the untransformed parent line. The transformed potato plants further contain the npt II gene from Escherichia coli under the control of the Nos promotor and terminator from Agrobacterium tumesfaciens. The npt II gene was introduced as a selection marker to facilitate the isolation of transgenic plants during the transformation process. Plant cells expressing the npt II gene show an increased resistance to the antibiotic Kanamycin as compared to plant cells without npt II expression.


6. Brief description of the method used for the genetic modification:
The binary vectors pBinARHyg-AtSDD1 or pBinAR-StSDD1 were constructed based on the Bin19 derivatives pBinARHyg or pBinAR, respectively. The resulting vectors were used to transform Agrobacterium tumefaciens strain C58C1. Axenic leaf cuttings of Solanum tuberosum cv. Desiree were incubated for 3 to 5 minutes in a suspension of the respective genetically modified Agrobacteria. Afterwards, these leaf cuttings were incubated on a shoot induction media containing the antibiotics Hygromycin or Kanamycin, respectively, to select for transformed cells and the antibiotic Cefotaxim or Tricarcellin to distroy the Agrobacteria. Regenerating shoots were transferred to a medium containing Cefotaxim or Tricarcellin and cultivated for at least two passages under these 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 applicable

Experimental Release

1. Purpose of the release:
Under greenhouse conditions, the potato plants that were transformed with the sense construct of the SDD1 gene showed decreased stomata densities, those transformed with the RNAi construct of the SDD1 gene showed increased stomata densities. The experimental release will provide data how the stomata density correlates to the performance of the plant in a field environment, where the plants are subjected to the natural diurnal and seasonal changes in environmental, especially climatic parameters.

2. Geographical location of the site:
14476 Potsdam/Golm, Flur 1, Flurstück-Nr 955, Potsdam, Brandenburg, Germany

3. Size of the site (m2):
A maximum number of 1800 GM plants/year are planted on a 500 m² release site in a trial area of 77000 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 GM plant has not been released before.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
In Germany, potato plants have a very low dispersal range and do not survive outside agronomic environments. Potato does not hybridise with any species growing wild in Germany. The low survival rate in natural environments is mainly due to the very low frost resistance of any part of the plant except seeds. Frost resistance as a major limiting trait for the survival of potato depends on a several genes which to our present knowledge lack in the genom of the cultivated Solanum tuberosum. The alteration of the activity of a single potato enzyme in the transgenic plants is unlikely to increase the frost resistance.

Brief description of any measures taken for the management of risks:
Potato has a very low dispersal capacity and does not hybridise with any species growing wild in Germany. Thus, GM potato can be isolated in the release site (= the area that is planted with the GM potato) by keeping a minimum distance of 20 m between the GM potato and any potato cultivation that is not monitored as stated below. This requirement is met by placing the release site accordingly on the test site. The release site is monitored for volunteers during the growth season of the year following the release. Any potato volunteer on the release site are destroyed. The post-harvest survey is repeated until the number of potato volunteers in the respective release site is zero for one year.

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 information

Consent given by the Member State Competent Authority:
Yes
01/04/2005 00:00:00
Remarks: