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Notification report


General information

Notification Number
B/DE/03/155

Member State to which the notification was sent
Germany

Date of acknowledgement from the Member State Competent Authority
03/12/2003

Title of the Project
Deliberate release of genetically modified marker-free amylopectin-potatoes for verification of starch quality and stability of transferred marker-free sequences under practical field conditions

Proposed period of release:
01/05/2004 to 31/10/2013

Name of the Institute(s) or Company(ies)
Bavarian State Research Center for Agriculture, Institute for Crop Production and Plant Breeding, ;


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 tuberosumtuberosum

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Marker-free trait sequences comprising 581 base pairs (bp) of exon 2 of the granular bound starch synthase gene (GBSS) from potato inserted in anti-sense orientation under transcriptional control of 5'-regulatory GBSS sequences were transferred into potato using agrobacteria. Transcription of anti-sense GBSS is terminated by terminator sequences from Cauliflower Mosaic Virus (CaMV). Notably, selection genes have not been transferred. Anti-sense GBSS sequences lead to a modified starch composition in potato tubers and enrichment of amylopectin.

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 transferred genetic material (T-DNA of pMFlp9-1) is surrounded by right and left border sequences (RB, nopaline type, 77 bp; LB, octopine type, 216 bp) from pPCV002 to promote agrobacterial gene transfer. It contains 805 bp 5'-regulatory GBSS sequences for a tuber enriched expression, exon 1 (11bp) and intron 1 (220 bp) GBSS sequences as transcriptional enhancers followed by 581 bp GBSS exon 2 sequences in anti-sense orientation to inhibit GBSS and to modify the starch content of the tuber tissue thereby. 162 bp terminator sequences from CaMV 35S locus are included to terminate anti-sense GBSS transcription.

6. Brief description of the method used for the genetic modification:
A novel marker-free binary T-DNA vector pMFlp9-1 was constructed using standard molecular cloning methods. The binary plasmid pMFlp9-1 was transferred into agrobacteria (GV3101/pMP90RK) via electroporation. Internodial tissue from 4 weeks in vitro grown tip cultures of potato cultivar Walli was incubated with cell suspensions of GV3101/pMP90RK/pMFlp9-1. Regeneration of treated plant tissue was done using standard plant tissue culture techniques. Selection procedures were not performed. Over a period of 3 to 16 weeks after inoculation regenerated shoots were harvested. Agrobacteria were removed using Cefotaxim during regeneration and in vitro growth. Approximately 5000 plants were generated. In vitro induced mini tubers were treated with iodine solution. Line #1332 showed a modified starch content

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:
Deliberated release is performed (1) to verify results from in vitro cultures and green house grown plants of line #1332 concerning the modified starch content of marker-free asGBSS potatoes, (2) to analyse genetic stability and hypothetical epigenetic effects under field conditions (chase specific monitoring) and (3) to characterise modified starch from this plants using industrial parameter. Moreover, general monitoring (e.g. plant development), monitoring of volunteers from tubers and seeds, glycoalkaloid measurements and frequency of out crossing will be accomplished. Effects on bumblebee populations will be monitored.

2. Geographical location of the site:
Flur "Gereut"; 85354 Freising; Bavaria; Germany; Europe

3. Size of the site (m2):
7700 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 genetically modified plant #1332 has not been released before

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The genetically modified potato plant #1332 carries GBSS sequences from potato on agrobacterial T-DNA and does not harbour any antibiotic or herbicide selection gene. The introduced DNA does not encode new proteins, but inhibits endogenous GBSS leading to modified starch quality of the tubers. Therefore, selection advantages due to the genetic modification are not evident. Increased persistence of marker-free plants in agronomic regions or an enhanced invasivity in natural areas is not expected, as transferred sequences can be already found in potato. To our present knowledge, spreading antibiotic resistance genes and any other possible harmful effects on the environment can be excluded in deliberated release experiments with the marker-free potato line #1332. On the other hand, significant environmental benefits are expected during amylopectin production from such tubers. Large amounts of solvents and acids and a huge quantity of waste water will be avoided.

Brief description of any measures taken for the management of risks:
Principally, biological safety of potato is high for following reasons. Potato is not a native plant in Europe and does not pollinate any wild species in this area. Potato is self and insect pollinating having pollen with a very low dispersal range. No potato volunteers can be found outside agronomic regions. Nevertheless, a case specific monitoring towards stability of the transgene, a general monitoring according to good breeding practice, monitoring of volunteers of tubers or seeds and examinations to describe the glycoalkaloid content and out crossing frequency will be performed. Appropriate plant production techniques are applied (e.g. two rows of pollen acceptor plants, special soil treatment to minimise seed volunteers). In post-harvest inquiries any remaining potato volunteers on the release site will be removed, chopped and composted

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
A monitoring plan according to directive 2001/18/EC is performed. General and chase specific screens are foreseen. Among the most important features will be a classical description of line #1332 according to good breeding practice, examination of transgene stability of marker-free potatoes and their breeding products, quantification of the number of volunteers from tubers or seeds and inspection of glycoalkaloid content in tubers. Field trials are designed to monitor the frequency of out crossing.

Final report
-

European Commission administrative information

Consent given by the Member State Competent Authority:
No
19/05/2004 00:00:00
Remarks: