Deliberate release into the
environment of GMOs for any other purposes than placing on the
market
Genetically modified marker-free
amylopectin-potatoes for verification of starch quality and
stability of transferred marker-free sequences under practical
field conditions - Bavarian State Research Center for
Agriculture, Institute for Crop Production and Plant Breeding |
Date of publication: March 19, 2004
Source:
http://gmoinfo.jrc.it/gmp_browse_geninf.asp
Notification
report
General information
Notification Number:
B/DE/03/155
Member State:Germany
Date of Acknowledgement: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 From: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
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)
|
Common Name |
Family Name |
Genus |
Species |
Subspecies |
Cultivar/breeding line |
| potato
|
solanaceae
|
solanum
|
solanum
tuberosum |
tuberosum
|
|
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;
4. 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. |
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