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Notification of deliberate release and placing on the EU market
of GM organism: fungal resistant wheat, Germany 2004 (II),
Syngenta Seeds GmbH |
Date of publication: January 26, 2004
Source:
http://gmoinfo.jrc.it/gmp_browse_geninf.asp
Notification
report
General information
Notification Number:
B/DE/03/152
Member State: Germany
Date of Acknowledgement:14/10/2003
Title of the Project:
Fungal resistant wheat Germany 2004 (II)
Proposed period of release From:01/03/2004
To:31/10/2004
Name of the Institute(s) or Company(ies): Syngenta
Seeds GmbH;
3. Is the same GMPt release planned elsewhere in the
Community?
No
Planned outside the EU: USA 02-074-06n, 03-073-03n Canada
02-SYN1-163-WHT01, 03-SBI1-245
4 - Has the same GMPt been notified elsewhere by the same
notifier?
Yes
If yes, notification number(s):
-
Genetically
modified plant
1. Complete name of the
recipient or parental plant(s)
|
Common Name |
Family Name |
Genus |
Species |
Subspecies |
Cultivar/breeding line |
| spring
wheat |
poaceae
|
triticum
|
triticum
aestivum |
|
|
2. Description of the traits and characteristics which have
been introduced or modified, including marker genes and previous
modifications:
FRG: Gene of fungal origin conferring tolerance to Fusarium
pathogens.
PMI: Phosphomannose Isomerase gene isolated from Escherichia
coli, conferring tolerance to mannose.
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 plasmids used for the transformation were derived from
the pUC19 plasmid described by Yanisch-Perron et al (1985).
Plasmid contents:
pZMLR 14 (6610bp)
Promoter: Ubi
Gene: PMI
Terminator: 35S
Intron: Intron #9pepc (I-9)
Ampicillin resistance gene controlled by a prokaryotic promoter
sequence: Amp (bla gene).
pZMLR 69 (6636bp)
Promoter: Ubi
Gene: FRG
Terminator: nos
Ampicillin resistance gene controlled by a prokaryotic promoter
sequence: Amp (bla gene).
Nature and source of the DNA coding sequences within PZMLR 14
and PZMLR 69
- Ubi: promoter of a maize ubiquitin gene together with the 1st
exon and the 1st intron of the gene (Christensen et al, 1992).
- PMI: Phosphomannose Isomerase Gene isolated from Escherichia
coli (Miles and Guest,1984). This gene is used as a marker for
the transformation, and allows positive selection on mannose
(Bojsen et al 1994, Joersbo et al 1998, Reed 1999, Negrotto et
al 2000).
- Intron #9pepc (I-9): Intron isolated from the maize
phosphoenolpyruvate carboxylase gene (pep-c) (Hudspeth and
Grula, 1989); its role is to stimulate the expression of the PMI
gene.
- 35S: Termination sequence of the cauliflower mosaic virus
gene.
- Nos: Termination sequence of the nopaline synthase gene,
isolated from Agrobacterium tumefaciens (Depicker et al 1982,
Bevan et al 1983).
- FRG: cDNA isolated form a fungal species, coding for an enzyme
capable of conferring Fusarium tolerance.
Prior to gene delivery, these plasmids were cut using
restriction enzymes in order to eliminate the carrier fragment
of the bacterial marker that confers resistance to ampicillin
and to achieve transfer to the plant of only the plasmid
fragment that carries the necessary genetic material for
effective transformation.
Wheat plants with the same modification were evaluated in field
trials in the USA and Canada in 2002 and 2003.
6. Brief description of the method used for the genetic
modification:
Biolistics method.
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:
To compare the agronomic performance (pathogen infestation
level and mycotoxin level) of wheat modified to express an
enhanced resistance to Fusarium pathogens with existing
non-modified varieties, grown under standard agronomic
conditions in Germany.
Two different transformation events will be assessed, compared
with five conventional genotypes used as controls.
2. Geographical location of the site:
Location: Baalberge
Postcode: D-06408
District: Bernburg
Federal state: Saxony-Anhalt
Company:
Agrargenossenschaft Baalberge eG
Umgehungsstraße 30
D-06408 Baalberge
Saxony-Anhalt
3. Size of the site (m2):
- The total trial ground (surface of the trial plus stripped
area plus border of dicotyleton plants) will be approx. 8.300
m².
- The area comprising the released GM wheat plus the non GM
wheat border rows (no less than 5 m) will be approximately 450
m².
- The trial will be surrounded by a 5m area stripped of all
plants and a 30m border of dicotyleton plants.
- The total number of GM seeds to be used in this field trial
will not be more than 6.000.
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:
No adverse effects were noted from wheat plants with the same
genetic modification released in the field in the USA and Canada
in 2002 and 2003.
Environmental
Impact and Risk Management
Summary of the potential
environmental impact from the release of the GMPts:
This application concerns a small-scale research trial which
will take place in Baalberge, Germany. Due to the
characteristics of the wheat plants, effects are expected to be
limited to the site of the trial, and to be transient in nature.
Wheat is an annual plant that reproduces by means of seed. Seeds
that are shed on the soil before or during harvest may
overwinter and sprout the following spring. However seed is
unlikely to survive longer than 2 years due to lack of dormancy
(Field Agronomist, personal communication) and volunteers are
easily controlled under standard agricultural conditions.
Wheat flowers display characteristics that do not favour
cross-pollination. Under natural circumstances the pollination
of wheat relies mainly on self-pollination (average level of
97-99%). Compared to other allogamous grasses, the production of
pollen by a wheat ear is very limited, 10% and 2.5% compared to
that of rye and maize inflorescence respectively (de Vries,
1971).
Wheat pollen is dispersed by the wind (David & Pham, 1993) but
it is fairly heavy, which restricts its dispersal distance (de
Vries, 1971 and 1974, Anand and Sharma, 1974).
Wheat pollen is highly sensitive to environmental conditions.
Pollen viability for periods of time ranging from 1 min to
approximately 30 min in optimal field conditions of 20°C, 60%
relative humidity have been quoted (Poehlman 1987).
Whilst wheat species may be present at the release site, they
will be at least 50 metres away from the transgenic trial site.
These crops are being grown for research and development
purposes only. There is potential for wheat, rye and triticale
crops to be planted in areas surrounding the field, but at a
distance of at least 100 metres from the trial.
There are a small number of Aegilops species that have the
potential to hybridise with wheat in Germany under natural
conditions. All species of Aegilops are annuals, often growing
in dry, open habitats, not on managed agricultural land. The
species described are most prevalent in Southern Europe, and are
not indigenous in Germany.
A survey of the flora surrounding the field site confirmed that
sexually compatible wild species are not present in the
experimental fields.
In summary, the self pollinating nature of wheat, the short
pollen life, the absence of wild relative species with which
hybridisation is possible at the site, the 5m border of non
modified wheat and the distance the trial will be located from
other wheat crops, indicates the likelihood of pollen flow
resulting in successful cross-pollination of the same or
sexually compatible species, or from the same or other sexually
compatible species to the trial plot is minimal, under the
conditions of the trial.
Any ungerminated seed remaining after sowing or produced as a
result of the trial may emerge as volunteers. To help with
volunteer control, any residual grain will be left on the
surface of the trial plot and encouraged to germinate by rain or
irrigation. The following year, a crop other than cereal that
will allow the clear identification of volunteers will be grown
and any wheat volunteers emerging will be removed and
incinerated or destroyed by treatment with an appropriate
herbicide. The site will be monitored for at least a year after
harvest.
The genetic modifications described above are unlikely to confer
any invasiveness characteristics to the plant since the intended
phenotypic expression of the modification is an increase in
Fusarium tolerance. If the plants are host to the Fusarium
species targeted by the gene, the development of these species
on these plants could be reduced. This selective advantage is
only likely to be for the duration of the growing season since
wheat does not possess the agronomic qualities to enable it to
become a weed and cannot survive without human intervention.
In summary, the likelihood of the modified wheat plants becoming
more persistent or invasive than non-modified plants is
considered to be negligible.
Population levels of non-target organisms are likely to remain
unaffected in this trial because of the specificity of the
intended effect of the modification. No direct, indirect,
immediate or delayed effects have been noted in previous USA and
Canadian trials with the same genetically modified wheat.
Being expressed constitutively both proteins encoded by the
fungal resistance gene and the PMI gene may be present in the
soil as a result of processes such as root damage. However, the
FRG is isolated from a common fungal species which is widely
prevalent in nature and is likely to be present in plants that
are colonised with the fungal species.
Neither the PMI protein sequence nor the FRG protein sequence
show homology to known allergens or toxins when searched against
allergen/toxin databases.
The trial is a small-scale research trial, and all the plants
used in the release (GM or non-GM) will be destroyed at the end
of the trial (except those retained for specific laboratory
analysis) and will therefore not be used as human food or animal
feed.
Biogeochemical processes are not expected to be affected in any
way by the presence of the modified plants.
The cultivation and management techniques used for the modified
wheat are the same as those used in good agricultural practice.
Brief description of any measures taken for the management of
risks:
During the trial, the plot will be monitored on a regular
basis for volunteer plants. Any found will be destroyed. After
termination of the trial, the plot will be monitored for at
least a year. Any volunteers detected during the monitoring
period will be destroyed. If during the monitoring procedure,
concerns arise as to the persistency of the plants the
monitoring period will be extended. A crop other than cereal
that will allow the clear identification of volunteers will be
grown in the following season and any wheat volunteers emerging
will be removed and incinerated or destroyed by treatment with
an appropriate herbicide.
If necessary the wheat plants can be easily destroyed using a
herbicide. In the event of small-scale vandalism, individual
plants uprooted or damaged will be removed and destroyed by
incineration.
If practicable, without compromising the data that would have
been obtained, the plants may be replaced and /or replanted.
In the event of large-scale vandalism where it is deemed the
trial cannot continue, the trial will be terminated by the
application of a herbicide and the plant material will be
disposed of as described above.
Summary of foreseen field trial studies focused to gain new
data on environmental and human health impact from the release:
The planned trial is a research field trial which aims to
compare the agronomic performance of wheat modified to express
an enhanced resistance to Fusarium pathogens with existing
non-modified varieties, grown under standard agronomic
conditions in Germany. It is not specifically designed to gain
new data on the environmental and human health impact of the
release.
Additional information:
REFERENCES :
- Anand S C and Sharma S C (1974). Pollen dispersal in wheat
(Triticum aestivum). Wheat Information Service, 1974, 38: 27-28.
http://www.grs.nig.ac.jp/wheat/wis/No38/p27/1.html
- Bevan M W, Barnes W M, Chilton M D (1983). Structure and
transposition of the nopaline synthase gene region of T-DNA.
Nucl. Acids Research 11 369-385
- Bojsen K I, Donaldson A, Haldrup M, Joersboe J, Kreiberg J,
Nielsen F, Okkels and Petersen S (1994). Mannose or Xylose based
positive selection Patent WO 94/20627 1994.
- Christensen, AH Sharrock, RA and Quail, PH (1992). Maize
polyubiquitin genes - structure, thermal perturbation of
expression and transcript splicing, and promoter activity
following transfer to protoplasts by electroporation. Plant
Molecular Biology 18 675-689.
- David J L & Pham J L (1993). Rapid changes in pollen
production in experimental outcrossing populations of wheat. J.
Evol. Biol. 6 659-676
- Depicker A et al. (1982) "Nopaline synthase: transcript
mapping and DNA sequence". J. Mol. Appl. Genet. 1 561-573
- de Vries A P (1971). Flowering biology of wheat, particularly
in view of hybrid seed production - a review. Euphytica 20
152-170
- de Vries A P (1974). Some aspect of cross-pollination in wheat
(Triticum aestivum L;) 4. Seed set on male sterile plants as
influenced by distance from the pollen source, pollinator : male
sterile ratio and width of the male sterile strip. Euphytica 23:
601-622
- Hudspeth R L and Grula J W (1989). Structure and expression of
the maize gene encoding the phosphoenolpyruvate carboxylase
isozyme involved in C4 photosynthesis. Plant Molecular Biology
12 579-589
- Joersbo M, Donaldson I, Kreiberg J, Petersen S G, Brunstedt J,
and Okkels F T, (1998). Analysis of mannose selection used for
transformation of sugarbeet. Molecular Breeding 4 111-117.
- Miles J S and Guest J R (1984). Nucleotide sequence and
transcriptional start point of the phosphomannose isomerase gene
(man A) of Escherichia coli. Gene 32 41-48.
- Negrotto D, Jolley M, Beer S, Wenck A R, and Hansen G (2000).
The use of phosphomannose-isomerase as a selectable marker to
recover transgenic maize plants (Zea mays L.) via Agrobacterium
transformation. Plant Cell Reports 19 798-803.
- Poehlman J M (1987). Breeding Wheat and Triticale, p. 220-239,
In J. M. Poehlman, ed. Breeding Field Crop. AVI Publishing
Company Inc., Westport.
- Reed J N, Chang Y F, McNamara D D, Beer S and Miles P J
(1999). High frequency transformation of wheat with the
selectable marker mannose-6-phosphate isomerase (PMI). In Vitro
35 57-A abstract P-1079.
- Yanisch-Perron C, Viera J and Messing J (1985). Improved M13
phage cloning vectors and host strains; nucleotide sequence of
M13mp18 and pUC19 vectors. Gene 33 103-109. |
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