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Notification report


General information

Notification Number
B/ES/13/20

Member State to which the notification was sent
Spain

Date of acknowledgement from the Member State Competent Authority
15/04/2013

Title of the Project
Wheat variety trial with a very low content of celiac-toxic epitopes.

Proposed period of release:
01/12/2013 to 31/07/2014

Name of the Institute(s) or Company(ies)
Instituto de Agricultura Sostenible, CSIC, ;


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
wheatpoaceaetriticumtriticum aestivumBobwhite

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
We have silenced the gliadins in wheat grain by using a hpRNA vector containing an endosperm specific promoter, a sense fragment of a gliadin sequence, the intron 1 of the maize ubiquitin, an antisense fragment of a gliadin sequence and the nopaline synthase terminator from Agrobacterium tumefaciens.
The pAHC25 plasmid containing the selector gene bar and the uidA marker gene (GUS) has also been introduced together with the former plasmid.


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:
We have inserted the following plasmids:

A) pDhp_omega_alpha plasmid containing the following elements:
1) D-hordein promoter, derived from Hordeum chilense, its function is to drive the expression of the RNAi construct in wheat endosperm.
2) A sequence of 361 base pairs corresponding to wheat gliadins sense orientation, from bread wheat, its function is to pair with sequence 4)
3) Intron 1 of the maize ubiquitin from maize, its function is to loop between sequences 2) and 4)
4) A sequence of 361 base pairs corresponding to wheat gliadins in antisense orientation, from bread wheat, its function is to pair with the sequence 2)
5) NOS termination signals, derived from Agrobacterium tumefaciens, its function is to signal the end of transcription
6) pUC18 vector sequences, used for cloning of the different elements, contains the ampicillin resistance gene. Without any function in the plant.

B) Plasmid pAHC25 which contains the following elements:
1) The ubiquitin promoter from maize, its function is to drive the expression of the bar gene.
2) Ubi1 intron of the maize ubiquitin from maize, its function is to enhance the expression of the bar gene.
3) bar gene derived from Streptomyces hygroscopicus, its function is as selector gene.
4) NOS termination signals, derived from Agrobacterium tumefaciens, its function is to signal the end of transcription.
5) The ubiquitin promoter from maize, its function is to drive the expression of the gene uidA (GUS).
6) Ubi1 intron of the maize ubiquitin comes from maize, its function is to enhance the expression of the gene uidA (GUS).
7) The uidA gene (GUS), derived from E. Coli, its function is of marker gene.
8) NOS termination signals, derived from Agrobacterium tumefaciens, its function is to signal the end of transcription.
9) pUC18 vector sequences, used for cloning of the different elements, containing the ampicillin resistance gene. Without any function in the plant.


6. Brief description of the method used for the genetic modification:
An equimolar mixture of the two plasmids described in section 4 has been introduced into wheat cells by acceleration with microparticles. After the in vitro selection using PPT as selective agent, plants containing the plasmids pDhp_omega_alpha and pAHC25 were regenerated.

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:
The release aims the production of 500 kg of grain for a clinical trial with the wheat flour in Reina Sofía hospital of Córdoba. The wheat line, which is intended to test, has low reactivity in relation to celiac disease shown by T cell test previously performed (PNAS, 2010. 107:17023-17028), and the results of monoclonal antibodies currently available. The flour from the field trial will be used to conduct the clinical trial with celiac patients. The clinical trial will be designed by Nutrition and Internal medicine Department of the above mentioned hospital.

2. Geographical location of the site:
The plot, where we intend to grow genetically modified wheat, is located on the site FUENTE PALMERA (CÓRDOBA).

3. Size of the site (m2):
An area of approximately 1000 m² within this plot will be sown with the genetically modified wheat.

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 previous release of this GM-plant has been carried out.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The purpose of the genetic modification was silence gliadins genes in wheat grain, responsible of celiac disease, using RNA interference techniques. The genetic modification inserted, silencing of gliadins in the grain, is not expected to confer an selective advantage and/or disadvantage in natural environments. Not expected environmental benefits.

Brief description of any measures taken for the management of risks:
The release site will be prepared and handled in accordance with good wheat trial practices. No wheat or compatible species will be planted at distances less than 200 m of the trial. A 2 m band, free from grass will surround the trial field. At the end of the trial, a broad-spectrum herbicide will be applied on the trial area. The plant material remaining after herbicide application will be crushed and incorporated into the soil to a depth of 20 cm. In the following year, a non-cereal crop will be cultivated, allowing clear identification of wheat plants
In the case of wheat field being damaged by natural causes before and / after flowering specific actions will be taken.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Not applicable

Final report


European Commission administrative information

Consent given by the Member State Competent Authority:
Yes
12/11/2013 00:00:00
Remarks: