Notification report

General information

Notification Number: B/DE/03/150

Member State:Germany

Date of Acknowledgement:29/08/2003

Title of the Project:
Effect of the leghemoglobin gene on the carbohydrate metabolism in transgenic plants

Proposed period of release From:01/03/2004 To:31/12/2008

Name of the Institute(s) or Company(ies): Max-Planck-Gesellschaft represented by the Max-Planck-Institut für Molekulare Pflanzenphysiologie;

3. Is the same GMPt release planned elsewhere in the Community?
No

4 - Has the same GMPt been notified elsewhere by the same notifier?
No

Genetically modified plant

1. Complete 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:
- Leghemoglobin gene from Lotus japonicus in sense orientation to the promoter of the patatin gene B33
- 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 partART27.

Genetic modification

3. Type of genetic modification:
Insertion;

4. In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
- Potato plants were transformed with a plasmid containing the leghemoglobin gene from Lotus japonicus in sense orientation to the promoter of the patatin gene B33 from Solanum tuberosum, terminated by the OCS terminator from Agrobacterium tumefaciens and the npt II gene from Escherichia coli under the control of the Nos promotor and terminator from Agrobacterium tumesfaciens. The expression of the leghemoglobin gene in the tubers of potato is assumed to affect the oxygen transport and the redox regulation in the tuber. Under greenhouse conditions, the insertion of the leghemoglobin had a positive effect on the starch content of the tubers. 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 B33-Leghemoglobin-OCS gene was cloned in the binary vector pART27. The resulting vector was 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 these genetically modified Agrobacteria. Afterwards, these leaf cuttings were incubated on a shoot induction media containing the antibiotics Kanamycin 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 leghemoglobin gene show an increased starch content as compared to the untransformed parental cultivar. The experimental release will provide data whether this valuable trait is stable under field conditions.

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

3. Size of the site (m2):
Total area of the test site: 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. The trait that was added in GM plants, the leghemoglobin gene, can affect the redox regulation system of the cell and thus alter a wide range of processes in the plant. However, 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.

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 has been planted with the GM potato) by keeping a minimum distance of 20 m between the GM potato and any other potato cultivation. This requirement is met by the 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 reduced to less than two plants per ha.

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
not applicable