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Notification report


General information

Notification Number
B/ES/06/41

Member State to which the notification was sent
Spain

Date of acknowledgement from the Member State Competent Authority
21/02/2006

Title of the Project
Introgression to the red rice weed.

Proposed period of release:
01/05/2006 to 30/10/2006

Name of the Institute(s) or Company(ies)
IRTA (Institut de Recerca i Tecnologia Agroalimentàries), Passeig de Gràcia 44. Barcelona.Spain;


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

Has the same GMPt been notified elsewhere by the same notifier?
Yes

If yes, notification number(s):
B/ES/03/37; B/ES/05/24;

Genetically modified plant

Complete name of the recipient or parental plant(s)
Common NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
ricepoaceaeoryzaoryza sativaSenia

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
One transgenic line (named G9-bar) containing the bar gene, conferring resistance to the herbicide ammonium glifosinate will be used.

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:
Line G9-bar: pUbi:bar:tnos

The source and function of each constituent fragments are:

pUbi: function: ubiquitin constitutive promoter (Christensen et al., 1992)
Source: Zea mays

tNos: function: nopaline-syntase terminator.
Source: pTiT37 plasmid from Agrobacterium tumefaciens.

lacZ alpha: function: beta-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


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. The vector used was the pCambia 3300.

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:
In previous studies, we demonstrated that hybridization between transgenic and red rice takes place in some extent (Messeguer et al., 2004) 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.

This field trial is 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 in combination with three 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 on 2004 the line S-bar-gus was planted. On 2005, line S-bar-gfp was planted in 2/3 parts of the plot whereas this year, only the 1/3 of the plot will be planted with the S-bar line, named G9-bar. 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.

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.

Studies carried out in greenhouse conditions have shown that introduced gene 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):
Four plots of 5 x 10 m2 with 16 transgenic plants/m2. Each one of these plots will be surrounded by non transgenic Senia cv occupying a total surface of 550 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:
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, the field trial planned here will allows us to evaluate how different agricultural practices may control the effective introgression of transgenes into the red rice.


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. 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 15 m 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. Weekly controls of agronomic traits will be performed.

At the end of culture, seed samples will be harvested manually. The rest of seed will be burned. The vegetal parts incorporated in the soil. The plot will be monitoring for re-growth and eventual volunteer plants will be eradicated 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
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European Commission administrative information

Consent given by the Member State Competent Authority:
Yes
12/04/2006 00:00:00
Remarks:
The competent authority for give the consent of these field trials is the Autonomous Community of Cataluña.