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Notification report


General information

Notification Number
B/RO/17/01


This notification has been withdrawn by the notifier
14/06/2017


Member State to which the notification was sent
Romania

Date of acknowledgement from the Member State Competent Authority
03/05/2017

Title of the Project
Using SUSIBA2 transgenic rice as a tool for creating a new genetically modified variety in sustainable system, characterized by large production capacity and reduced methane emissions.

Proposed period of release:
01/10/2017 to 31/12/2020

Name of the Institute(s) or Company(ies)
UASVMB - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of Biotechnologies, 59 Mărăşti Boulevard, District 1, Bucharest, Zip code 11464, Phone:004 021 224 28 93; Fax: 021 22 42 893;


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
ricepoaceaeoryzaoryza sativa

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
SUSIBA2 form of genetically modified rice, obtained in laboratories of the University of Stockholm, Faculty of Agriculture and Sciences, Uppsala, is characterized by large production capacity and reduced methane emissions. SUSIBA2 rice was mainly obtained by inserting into the genome of the Nipponbare rice variety, of a barley gene involved in the control of gene expression in plants. "Sugar Signaling in Barley 2" (SUSIBA2) is a transcription factor isolated from barley that regulate the expression of plant genes induced by sugars, its over-expression leading to greater accumulation of starch and obtaining larger amounts of biomass. Also, by over-expression of the gene encoding the transcription factor SUSIBA2, a redirection of the carbon source toward the stem, leaves and grains takes place and less toward the root system; thus, the microorganisms in the substrate in which rice grows, responsible for producing methane, have no access to the carbon source required for a normal metabolism, represented by exudates of nutrients in the rhizosphere and as a result, the level of methane synthesized and delivered in such conditions will be much lower.
The T-DNA region of the vector contains, besides the coding sequences of SUSIBA2 gene from barley, the hptII gene for hygromycin resistance and the gus gene for beta-glucuronidase enzyme synthesis, as well as sites for the restriction enzymes SacI, HindIII si SpeI.


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:
About 2 kb of SUSIBA2 gene from barley were inserted in the vector and transferred to the Nipponbare rice variety with the intention that after expressing of the transferred genetic material, the carbon flow to be redirected from the root system toward the aerial part. This phenomenon is taking place in SUSIBA2 transformed rice, because the gene transferred from barley is encoding a transcription factor involved in carbohydrate signaling, in metabolism and in the synthesis of starch.

Summary of the inserted DNA fragment inherited from SUSIBA2:

Genetic Element: Size (kb). Function.

- HvSUSIBA2- Sequence of aprox.2 kb (nucleotide sequence 247–2067) that is coding the gene SUSIBA2 (“sugar signaling in barley”) for a transcription factor implied in starch biosynthesis from barley (Su și colab., 2015)
- pHvSBEIIb - Nucleotide sequence 1–1010 from the promoter SBEIIb from barley (Su și colab., 2015)
- gus - Gene for β-glucunoridase enzyme from Escherichia coli; reporter gene from vector pCAMBIA 1301
- hptII - Gene for hygromycin phosphotransferase; marker gene of selection of the transformed clones, originating from the vector pCAMBIA 1301
- 35S - Promoter of 35S genes from the cauliflower mosaic virus (CaMV) – ensures the expression of marker/reporter genes
- nos - The terminal sequence 3’ of nopaline synthase gene originating from ADN-T of Agrobacterium tumefaciens, which determines the end of transcription and polyadenilation of ARNm; is originating from the vector pCAMBIA 1301.


6. Brief description of the method used for the genetic modification:
SUSIBA2 rice was obtained by genetic transformation of immature embryos of the conventional Nipponbare rice variety, by an Agrobacterium-mediated process. This transformation resulted in the stable integration of the T-DNA region of the binary vector pCAMBIA1301 into the genome of rice. T-DNA region of the vector contains the coding sequences of the gene SUSIBA2 from barley, hptII gene for hygromycin resistance and gus gene for the synthesis of beta-glucuronidase enzyme, as well as sites for the restriction enzymes SacI, HindIII and SpeI.

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 deliberate release of genetically modified rice in the environment for tests in greenhouse conditions has the goal of obtaining information on the volume of gaseous emissions and on the biological performance of genetically modified rice plants following the transfer of "sugar signaling in barley 2" gene (SUSIBA2) from barley to rice. The aim is, that based on the data recorded by testing this biological material, to obtain genetically non-modified rice, but having the qualities of SUSIBA2 rice as regards the reducing the greenhouse gas emissions and the high productivity.

2. Geographical location of the site:
Hortinvest greenhouse – UASVM of Bucharest, Romania.

3. Size of the site (m2):
Testing area 50 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:
Following the observations carried out in experimental fields during 2012-2014 in Fuzhou, Guangzhou and Nanning, China, and in phytotron conditions in Sweden on SUSIBA2 rice cultivated over several years, no adverse effects, immediate and / or delayed, have been reported on the environment or on human health.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
Since the tests will be carried out into protected and isolated space, in the greenhouse and vegetative mass will be destroyed at the conclusion of the experiment, we do not anticipate possible immediate and / or delayed effects on the environment or on human health.
The experimental data resulting from testing conducted to date in China and Sweden have concluded that the gene "Sugar Signaling in Barley 2" transferred from barley to rice, favors the allocation of carbon to biomass (seeds and stems), resulting in a high starch content in these tissues; on the contrary, the flow of carbon into the root system is reduced, resulting in a pronounced reduction in root exudates of nutrients, which leads to a suppression of methanogenesis. Consequently, cultivation of SUSIBA2 rice was associated with a reduction in methane in the rhizosphere and of methane emissions into the atmosphere. As a result, less gas emissions in the atmosphere of SUSIBA2 rice, is a characteristic with agronomic and ecological relevance for rice crops, which means in the same time a positive consequence for the environment.
Therefore, the tests to be carried out in this project in greenhouse conditions, in Romania, aim at getting new information on the biology, agronomic and phenotypic performance and the behavior of SUSIBA2 rice plants in our experimental conditions.


Brief description of any measures taken for the management of risks:
Monitoring and control measures that will be taken in order to minimize the potential risks associated with the introduction into the environment of genetically modified rice for testing, will include among others:
- Experiments with the test material will be placed in greenhouse compartments isolated from other experiments with plants;

- Sowing / planting and the daily monitoring of SUSIBA2 rice will be carried out only by the specialists included in the project, who will be trained on all precautions measures, according to the specific working protocols on handling with genetically modified material;

- Experimental variants will be monitored throughout the entire testing period, by performing, besides biometric measurements, also the samplings of methane emissions in the atmosphere;

- Biological material obtained, seeds and plant material, will not enter the food and feed chain; thus, the remaining biomass, seeds and any other wastes resulting from the experimental work will be destroyed by cutting in small pieces, autoclaving and incorporating in soil;

- seed samples, that will be send for analysis to the project coordinator, will meet the requirements for labeling and transport

At the end of the test period the notifier will submit a report to the competent authority; the report will detail the possible adverse effects on the environment, if they will occur.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
In Romania, as tests with the SUSIBA2 transgenic rice will be performed in the greenhouse, no adverse effects on the environment or human health, are expected.
However, any unanticipated adverse effects on human health or the environment would be reported immediately to the Competent Authority.


Final report
-

European Commission administrative information

Consent given by the Member State Competent Authority:
Not known