Notification report

General information

Notification Number

Member State to which the notification was sent

Date of acknowledgement from the Member State Competent Authority

Title of the Project

Proposed period of release:
01/05/2012 to 30/09/2012

Name of the Institute(s) or Company(ies)
Public University of Navarre, (Spain);

3. Is the same GMPt release planned elsewhere in the Community?

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

If yes, notification number(s):

Genetically modified plant

Complete name of the recipient or parental plant(s)
Common NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
tobaccosolanaceaenicotiananicotiana tabacumtabacumVirginia Gold and Havana-503

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Introduction of the nuclear gene thiorredoxin f of tobacco in the plastid genome of the same species. The marker gene is the aadA from Escherichia coli that confers resistance to the antibiotics spectinomycin and streptomycin.

Genetic modification

3. Type of genetic modification:

In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
Thiorredoxin f: nuclear tobacco gene coding for thiorredoxin f protein that is localized in the plastid.
PrrnG10L: promoter of the rrn plastid gene fused to the leader region of the gene10 of the bacteriophage T7.
3’rps16: 3’-region of the plastidial gene rps16.
aadA: aadA marker gene from Escherichia coli that confers resistance to the antibiotics spectinomycin and streptomycin.
Prrn: promoter of the plastidial gene rrn.
TpsbA: 3’-region of the plastidial gene psbA.
lox: two small sequences flanking the aadA gene. They allow the elimination of the marker gene by the exogenous recombinase Cre.

6. Brief description of the method used for the genetic modification:
Chloroplast transformation of tobacco leaves by the biolistic method (Svab Z, Hajdukiewicz P, Maliga P. 1990. Stable transformation of plastids in
higher plants. Proc. Natl. Acad. Sci. USA, 87:8526–30).

Experimental Release

1. Purpose of the release:
The purpose of the release is to verify that (similar to previous results obtained in greenhouse) transgenic plants expressing the thiorredoxin f in the chloroplasts differ from parental plants only in the endogenous levels of starch and soluble sugars.

2. Geographical location of the site:
Genetic modified tobacco plants will be released in Sartaguda (Navarra-Spain). The Experimental Field, which belongs to the Instituto Técnico de Gestión Agrícola (ITGA), a governmental Institution, is located in a dry area (5-50 mm precipitation) in south Navarra.

3. Size of the site (m2):
The experimental field has 19 ha. However we will be using only 150 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:
Not negative effects of GMOs over adventitious flora (weeds) or pests were detected. Weeds present in the experimental field equally affected wild-type and transgenic plants.
Health problems (allergic or toxic reactions) were not detected in workers handling the plants.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The introduced traits -expression in the chloroplast genome of the gene thiorredoxin f that is already present in the nuclear genome- are not expected to confer any selective advantage either any effect on environment when compared with control plants.

Brief description of any measures taken for the management of risks:
Despite the maternal inheritance of plastids (no transmission of transgenes via pollen) the release will be located in an area where tobacco is not commercially cultivated.
The experimental field is fenced and entrance is not allowed to nobody other people’s.
Flowers will be cut manually to avoid plants to set seeds.
Elimination of seeds: after harvesting all plants, the field will be watered and volunteer plants will be treated with herbicide.
The following year, tobacco will not be cultivated in this experimental plot. The plot will be monitored and the putative developed tobacco plants will be destroyed.

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
A total of 180 plants will be cultivated. Two different transgenic lines corresponding to the transformation event previously described, one for the var. Virginia Gold, the other of the var. Havana 503. Forty-five plants for each variety, divided in three plots of 15 plants each, will be cultivated. The size of the basic plot is 2,7 x 3 m. In addition, non-transformed plants will surround the transgenic plants.

Final report

European Commission administrative information

Consent given by the Member State Competent Authority:
20/04/2012 00:00:00