Deliberate release into the E.U.
environment of GMOs for any other purposes than placing on the market:
Evaluation of isolation distances and containment barriers to
control the gene flow between transgenic and non- transgenic
rice plants from the same cultivars. Introgression to the red
rice weed. - Institut de Recerca i Tecnologia Agroalimentàries,
Spain |
Date of publication: September 13,
2005
Source:
http://gmoinfo.jrc.it/gmp_browse_geninf.asp
Notification number:
B/ES/05/24 Member
State: Spain
Date of Acknowledgement: 01/05/2005
Title of the Project:
Evaluation of isolation distances and containment barriers to
control the gene flow between transgenic and non- transgenic
rice plants from the same cultivars. Introgression to the red
rice weed.
Proposed period of release From: 01/05/2005
To: 30/10/2005
Name of the Institute(s) or Company(ies): IRTA
(Institut de Recerca i Tecnologia Agroalimentàries);
3. Is the same GMPt release planned elsewhere in the
Community?
No
4 - Has the same GMPt been notified elsewhere by the same
notifier?
Yes
If yes, notification number(s):
B/ES/03/37
Genetically
modified plant
1. Complete name of the
recipient or parental plant(s)
Common Name
|
Family Name
|
Genus |
Species
|
Subspecies
|
Cultivar/breeding line
|
rice
|
poaceae |
oryza
|
oryza
sativa |
|
|
2. Description of the traits and characteristics which have
been introduced or modified, including marker genes and previous
modifications:
Three transgenic lines containing the bar gene, conferring
resistance to the herbicide ammonium glifosinate will be used.
These lines are: line S-1B containing bar and gusA genes; line
G9 bar-gfp, containing bar, gusA and hptII genes and line
G9-bar, containing only the bar gene.
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:
All lines were obtained by using the Agrobacterium mediated
transformation technique. The T-DNA contains:
Line S 1B : p35S:bar:t35S::p35S:gusA:tnos (This line has been
used in previous field trials (B7ES/00/07 and B/ES/01/07)
Line G9-bar: pUbi:bar:tnos
Line G9-bar-gfp (two T-DNAs inserted in the same
loci):pUbi:bar:nos and pgos2:gfp: trbsc::p35S:hptII:t35S.
The source and function of each constituent fragments are:
p35S: function: promoter from ARN 35S from CaMV
Source: cauliflower mosaic virus.
pgos2: function: promoter from gos2 rice gene (de Pater et al.,
1992).
Source: Oryza sativa
pUbi: function: ubiquitin constitutive promoter (Christensen et
al., 1992)
Source: Zea mays
tNos: function: nopaline-syntase terminator.
Source: pTiT37 plasmid from Agrobacterium tumefaciens.
trbcS: function: terminator from rice rbcS gene (Matsuoka et
al., 1988)
Source: Oryza sativa
t35S: function: terminator from CaMV ARN 35S
Source: cauliflower mosaic virus.
lacZ alpha:function: -galactosidase codifying sequence. It is
not express in plants, only in bacteria.
Source: Escherichia coli
bar: function: phosfinotricine acetyl transferase codifying
sequence.
Source: Stretomyces hygroscopicus
gusA: function: codifying sequence of -glucuronidase.
Source: Escherichia coli (Jefferson et al. 1987). Modified by
CAMBIA by introducing an intron to avoid its expression in plant
tissues (Ohta et al., 1990).
hptII Function: hygromycin phosfotransferase II codifying
sequence.
Source E. Coli (Gritz y Davies, 1983).
GFPS65T: Function: codifying sequence from the mutant protein
GFP: Ser 65Thr .
Source: gene gfp from Aequorea victoria (Heim et al, 1995).
6. Brief description of the method used for the genetic
modification:
Rice Senia cv was modified by using the Agrobacterium
mediated transformation technique according to Pons et al, 2000.
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:
There are two different field trials:
Field trial A: aimed at establishing the security distances
between transgenic and non-transgenic plants and to study the
efficiency of containment barriers on controlling the gene flow.
Transgenic lines with different markers will be placed at
different distances from non-transgenic plants in such a way
that cross-pollination inside and outside the field will be
monitored by analyzing the samples of seeds collected at the end
of the culture in different places and taking into account the
wind directions. Moreover, a “natural barrier” (a row of maize
plants) will be placed between transgenic and non-transgenic
plants in one half of the field. This field trial will be
carried only during this year.
Field trial B: aimed at assess the introgression of transgenes
into red rice, the only one weed compatible with cultivated rice
in Europe. This field trial was planned for 3 years (2004-2006).
As it was described in the SNIF from B/ES/03/37, transgenic
plants with a different molecular marker will be used in each
year combination with four different agricultural practices, the
most commonly used by rice growers to control the red rice weed.
Moreover, the first year a known number of red rice plants,
equivalent to a high infestation level were planted among
transgenic plants. This strategy will allow, at the end of the
field trial, to evaluate the contribution of each year to the
final introgression produced and will give a valuable overview
of the effect of the different agricultural methods on red rice
control among the three years of culture. The plots were divided
in three sections in such a way that the first year only line
S-bar-gus was planted. This year, line S-bar-gfp will be planted
in 2/3 parts of the plot whereas the third year, only th1/3 of
the plot will be planted with the S-bar line. Non-transgenic
plants from the same variety will surround all these plots. At
the end of the growing season samples of red rice seeds from
each sub-parcel and from surrounding non-transgenic plants will
be analyzed to determine the hybridization and the introgression
rate.
As described in C.4, field trials carried out in the same place
(B/ES/99/16, B7ES/00/07 and B/ES/01/07) clearly demonstrated
that gene flow from transgenic to non-transgenic plants takes
place to some extent but strongly decreases as distance
increases from 1 to 10m and there is a clear influence of the
dominant wind. Nevertheless, results obtained with these field
trials -that had a circular design and little size- needs to be
confirmed in a commercial-size field, in order to be able to
establish some general regulations to be applied in rice growing
areas.
On the other hand, hybridization between transgenic and red rice
takes place (see C.4) but there are not enough data to establish
de degree of introgression that could take place when a proper
agricultural technique focused to the control of this weed is
applied.
Genes transferred to transgenic plants used in this release do
not represent any selective advantage in comparison with
non-transgenic plants, except in the use of ammonium glufosinate
herbicide. Nevertheless, this herbicide is not currently used in
rice crop. Safety of phosphinothricin acetyltransferase is well
known. In relation to the marker genes, gusA gene encodes for
-glucuronidase protein, widely present in nature and the gfp
protein is widely used in medical tests.
Studies carried out in greenhouse conditions have shown that
introduced genes do not change the dissemination ability of
transgenic plants and that its agricultural behavior is similar
to that of non-transgenic plants.
2. Geographical location of the site:
IRTA Experimental Station. (Amposta) Tarragona, Spain.
3. Size of the site (m2):
Essay A: determination of security distances and efficiency
of containment barriers: 1720 m2 (840 m2 with transgenic
plants).
Essay B: introgression to red rice: 2700 m2 ( 400 m2 with
transgenic plants).
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:
A circular field trials designs (B/ES/00/07 and B/ES/01/07)
were carried out to assess the frequency of pollen-mediated gene
flow from a transgenic rice line S 1B, harbouring the gusA gene
and the bar gene encoding respectively ß-glucuronidase and
phosphinothricin acetyl transferase as markers, to conventional
rice in the Spanish japonica cultivars Senia. Frequencies of
gene flow based on detection of herbicide resistant, GUS
positive seedlings among seed progenies of recipient plants and
averaged over all the wind directions were 0.086 ± 0.007.
However, a clear asymmetric distribution was observed with
pollination frequency favoured in plants placed under the local
dominant winds. Southern analyses confirmed the hemizygous
status and the origin of the transgenes in progenies of
surviving, GUS positive plants. Examination of the influence on
gene flow frequency of the distance from the transgenic source
to recipient plots of conventional rice planted at 1, 2, 5 and
10 m distance revealed a clear decrease with increasing distance
which was less dramatic under the dominant wind direction. The
precise determination of the local wind conditions at flowering
period and pollination day time appear of primary importance for
setting up suitable isolation distances.
The same field trials designs were used to evaluate the gene
flow to red rice placed at different distances from the
transgenic plants and how the wind could influence the gene flow
to red rice plants growing in the borders. Frequencies of gene
flow averaged over all the wind directions were 0.036 ± 0.006 %.
However, as in the case of conventional rice, a clear asymmetric
distribution was observed with pollination frequency favored in
plants placed under the local dominant winds. Nevertheless
within a commercial transgenic rice field the influence of the
wind appears a less determinant factor because red rice plants
usually will grow isolated or in patches surrounded by
transgenic plants and consequently can be pollinated by all of
them. On the other hand, the wind influence on cross-pollination
has to be taken into account for the plants growing in the
borders. This is a very essential question to consider because
the real introgression of the genes will be minimized inside the
field by the usual control practices tending to destroy the red
rice but the wild plants in the borders can act as reservoirs of
the transgenic characters. Moreover, although the gene flow
values are relatively low, the shattering and dormancy of the
red rice seeds, which ensure their persistence in the field,
lead into an undesirable effect of durability of the transferred
genes. In consequence, whether one wants to avoid gene flow to
the red rice, crop management has to be changed. In this sense,
further studies are needed to evaluate how different
agricultural practices may control the effective introgression
of transgenes into the red rice. These studies are needed to
design new methodological tools for assessing systemic effects
within the diversity of environmental systems in which GMOs may
be cultivated.
Environmental
Impact and Risk Management
Summary of the potential
environmental impact from the release of the GMPts:
Introduced genes could not confer an increased selective
advantage in natural environments to transgenic plants because
the herbicide ammonium glufosinate is not commonly use in rice
fields.
One of the most potential environmental impacts of the release
of the GMPts is the risk of transgene spread throughout
cross-pollination. As has been described in C.4, we have
quantified the gene flow by using circular designs. The
objective of the field trials proposed here is to confirm if it
is possible to minimize the gene flow by increasing security
distances or by establishing containment barriers. On the other
hand, it is very important to establish at what degree the
introgression of transgenes to the red rice takes place in field
conditions and to know if it can be controlled by the
agricultural practices commonly used in controlling this weed.
Brief description of any measures taken for the management of
risks:
To prevent out-crossing with neighboring rice fields the
trial will be located 15m from any conventional rice field. It
has to be taken into account that the security distance
recommended by plant breeders is of 10 m.
Field trial will be keep free of weeds with the exception of red
rice introduced in field trial B. Weekly controls of agronomic
traits will be performed.
At the end of culture samples of seed will be harvested
manually. The rest of seed will be burned. The vegetal parts
will be burned on the plot or shallow incorporated in the soil.
The plot will be monitoring for re-growth and eventual volunteer
plants will be eradicated or analysed depending on the field
trial. In case of an emergency the plants can be destroyed
mechanically or by applying herbicides.
Summary of foreseen field trial studies focused to gain new
data on environmental and human health impact from the release:
As described in C.4 and D, these field trials will contribute
to better knowledge on gene flow in rice and to establish proper
regulations to be applied in case transgenic rice plants could
be introduced in Europe.
Final report
European
Commission administrative information
Consent given by the Competent
Authority: Not Known |
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