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Notification report


General information

Notification Number
B/CZ/11/4

Member State to which the notification was sent
Czech Republic

Date of acknowledgement from the Member State Competent Authority
28/12/2011

Title of the Project
Transformation of flax aimed at fatty acid desaturase 2 (FAD2) gene inactivation by RNA interference

Proposed period of release:
15/04/2012 to 30/06/2018

Name of the Institute(s) or Company(ies)
Agritec, Research, Breeding & Services, Ltd., ;


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
flaxlinaceaelinumlinum usitatissimum

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Flax plants were transformed with a vector construct bearing expression cassette with two FAD2 gene (coding for type 2 fatty acids desaturase) segments modified in a way to employ RNA interference system for suppression of desaturase function, and due to it to incerase the content of oleic acid in linseed oil. Along with modified FAD2 gene the insert contains hpt selectable gene to enable selection of transgenic plants, uidA (GUS) reporter gene and AmpR gene to support plasmid construct selection in bacteria.

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:
Linseed plants were modified using Agrobacterium-based vector constructs bearing an expression cassette with two flax gene FAD2 (coding for type 2 fatty acids desaturase) complementary segments to induce post-transcriptional gene silencing based on the interference of particular RNA transcripts and thus to suppress the conversion of oleic acid (omega-9) to omega-6 and omega-3 fatty acids what results in a change of fatty acids composition of the seed oil (elevated level of oleic acid). To enable the in vitro selection of transgenic plants bacterial hpt gene from Escherichia coli coding for hygromycin B phosphotransferase (HPT) and conferring resistance to the antibiotic hygromycin B was included in the construct together with E. coli uidA gene for beta-glucuronidase (GUS) reporter to monitor the presence and expression of cassette in the host organism. Modified gene of the interest (FAD2-i), which consists of two complementary segments separated by a spacer sequence is driven by common constitutive CaMV 35S promoter of Cauliflower mosaic virus. Close to FAD2-i position in the plasmid is bacterial AmpR gene conferring resistance to the antibiotic ampicilin, enabling the construct selection.

6. Brief description of the method used for the genetic modification:
Co-cultivation of linseed explants with Agrobacterium strain GV3101 was used to deliver the corresponding part (T-DNA) of the plasmid construct pPCV812i/35S/FAD2i/35S/GUS to the plant genome.

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:
Field experiments are aimed at the verification of biological characteristics and breeding parameters of selected, glasshouse tested, transgenic flax (linseed) lines obtained by introducing of modified FAD2 gene to flax genome to enhance the content of oleic acid in seed oil. Forms differing from original cultivars in oil composition, content and/or other breeding traits will be selected for the next crop breeding programs. Possible influence of transgene on plant morphology and biological characteristics will be another aim of the field studies.

2. Geographical location of the site:
Vikyrovice, region Olomouc

3. Size of the site (m2):
max. 100 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:
The same type of GM plants has not been released in the Czech Republic yet but numerous transgenic lines obtained by method of insertion mutagenesis, and other lines modified by different genes of interest (bar gene – to enhance plant tolerance to the herbicide application, SPI2 gene variants for inhibitors of serine proteases to enhance the ability of flax to resist microbial pathogens and/or insect pests, genes for metallothionenine αHMT1A or short peptide CP to enhance the accumulation of heavy metals in flax plants and their parts - phytoremediation) have been field-tested since 1999 based on approvals of the Ministry of Environment of the Czech Republic. No adverse effects have been identified either to the environment or to human health.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
Detailed analysis of the characteristics of the transgenic flax lines and accepting environment has shown that the risk of potential adverse effects on human and animal health and the receiving environment, resulting from the planned field trials with the GM flax lines, is consistently negligible.

Brief description of any measures taken for the management of risks:
To avoid possible pollen transfer by honeybees and other insects the experimental field distance from the nearest cultivation area of unmodified flax will be at least 300 m. At least 3 m-wide separating distance from other crops will be kept to decrease dissemination of seeds and to monitor possible effects of the GM organism on field flora and fauna around the experimental plots too. Sowing and harvesting machinery will be cleaned on site to prevent the dispersal of GM seed. The harvested material and seed will be transported from the site in closed and labeled containers to the laboratories for analyses. The remaining seed which will not be used in following seasons will be inactivated as well as the vegetative plant material. The area will be controlled for volunteers for a period of 1 consecutive year. During the next 6 years there will be no conventional flax cultivation, which equals the conventional crop rotation practice. Volunteers will be treated using conventional agricultural practices. During the release, the project leader and trained personnel will monitor the trial site at defined intervals.

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
10/05/2012 00:00:00
Remarks: