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
Amplification and evaluation of a transgenic corn line with biofortified endosperm with three vitamins.

Proposed period of release:
25/04/2015 to 31/12/2015

Name of the Institute(s) or Company(ies)
University of Lleida, ;

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):
B/ES/12/38; B/ES/13/16; B/ES/14/04;

Other notifications
B/ES/12/38In 2011, Louisiana State University LSU1109.1 (USA)

Genetically modified plant

Complete name of the recipient or parental plant(s)
Common NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
maizepoaceaezeazea maysmaysM73W

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
We created elite inbred South African transgenic corn plants in which the levels of 3 vitamins were increased specifically in the endosperm through the simultaneous modification of 3 separate metabolic pathways. The transgenic kernels contained 169-fold the normal amount of β-carotene, 6-fold the normal amount of ascorbate, and double the normal amount of folate. Levels of engineered vitamins remained stable at least through to the T4 homozygous generation. This achievement, which vastly exceeds any realized thus far by conventional breeding alone, opens the way for the development of nutritionally complete cereals to benefit the world’s poorest people.

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:
The selectable marker bar and 4 genes/cDNAs encoding enzymes in the metabolic pathways for the vitamins beta-carotene, ascorbate, and folate.
To increase beta-carotene levels, we introduced corn (Zea mays) phytoene synthase (psy1) cDNA under the control of the wheat LMW glutenin promoter and the Pantoea ananatis (formerly Erwinia uredovora) crtI gene (encoding carotene desaturase) under the control of the barley D-hordein promoter.
To increase ascorbate levels we introduced rice dehydroascorbate reductase (dhar) cDNA.
To increase folate levels we introduced the E. coli folE gene encoding GTP cyclohydrolase (GCH1), both under the control of the barley D-hordein promoter.

6. Brief description of the method used for the genetic modification:
Corn Transformation. Corn plants (Zea mays L., cv. M37W) were grown in the greenhouse and growth room at 28/20 °C day/night temperature with a 10-h photoperiod and 60–90% relative humidity for the first 50 days, followed by maintenance at 21/18 °C day/night temperature with a 16-h photoperiod thereafter. Immature zygotic embryos were excised at 10–14 days after pollination and cultured on N6 medium. After a further 5 days, the embryos were transferred to N6 medium containing high osmoticum (0.2Mmannitol, 0.2M sorbitol) for 5–6 h before bombardment, then bombarded with 10 mg of coated gold particles, as previously described. Bombarded callus was selected on phosphinothricin-supplemented medium, and transgenic plantlets were regenerated and hardened off in soil. A population of _75 transgenic plants was regenerated and screened by genomic PCR to identify primary transformants containing all 5 input transgenes.

Molecular analysis. The genomic PCR was carried out by using sets of 3 primers for each gene, generating overlapping products. Transgene expression was verified by mRNA blot analysis, and one transgenic line (L-1, Carolight) was identified expressing 4 genes.

Independent transgenic events were identified and characterized by PCR (see below) and then selfpollinated to produce T1 seeds. Homozygous T2, T3 and subsequent T4 generations were derived through selfing.

The transgenic kernels contained 169-fold the normal amount of β-carotene, 6-fold the normal amount of ascorbate, and double the normal amount of folate when compared to the wild type M37W.

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 objective of this application is the amplification of transgenic corn line L1 (Carolight)that contains three vitamins for its agronomic evaluation and future development (same objective then B/ES/12/38 and B/ES/13/16). We will evaluate: Effects on pests and natural fauna, suitability for industrial processing, ability to feed (we attach accepted manuscript in chicken feeding trials) and evaluation of their combining ability for collection and subsequent development of new hybrid varieties, new aspects to evaluate is expected.

2. Geographical location of the site:
We plan only Lleida municipality.

3. Size of the site (m2):
500 m2 surrounded by 6 rows of conventional corn.

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:
Three transgenic trial with the same event were performed in the same location B/ES/12/38, B/ES/13/16 and B/ES/14/04. Plants were totally normal showing only the different phenotype and genotype as expected for the introduced transgenes, when compared to the control M37W.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
- Within the consecutives multiplication generations no morphological, physiological or agronomic differences were detected between transformed plants and their conventional counterparts except for the presence of the three vitamins.
- The genetic modification introduced in the corn plant consists on the incorporation of different genes involved in the synthesis of the three vitamins, in order to improve the nutritive properties of the grain. Any negative effect on human health is predicted, quite the opposite.
- Vitamin consumption does not pose any risk on the health of livestock fed with vitamin enriched grain.
- The genetic modification does not target any organism; therefore no toxic effects on other organisms within the ecosystem are predicted.
- They are no sexually compatible species (neither cultivated nor wild-type) in Europe, since the corn plant has its origins in Central America.
- As cultivated specie, corn varieties have been bred to germinate in the consecutive crop cycle when the appropriate environmental conditions are present. Their survival out of the crop is very unlikely since they are highly adapted to crop conditions. Moreover, in the region where the trial field would be placed, those seeds able to germinate after the harvest period die due to autumn and winter colds. Therefore, any selective advantage of the transformed plants over wild-type plants in natural environments is predicted.
- There are no predicted effects on the biogeochemical cycles since the newly synthesized vitamins are mineralized like most of other organic compounds.

Brief description of any measures taken for the management of risks:
The GMO panel of the European Food Security Agency has confirmed that the cultivated transgenic corn is as safe as its conventional counterpart regarding potential effects on human and animal health and on the environment, on the basis of years of experience in cultivating and evaluating transgenic corn.
In the case of this multivitamin transgenic corn the risk is even lower since it has been engineered to increase the nutritional value of the grain, with no aim to neutralize other organisms. Despite of this, however, the following safety measures will be implemented:
- The field will be placed in an urban zone where corn is not cultivated.
- The field for transgenic corn multiplication will be surrounded by at least 6 rows of an authorized conventional variety in order to decrease the amount of released pollen outside the growing area.
- The grain harvested from the corn in the edges of the field will be destroyed and buried.
- The grain harvested from the transgenic corn L1 will be manipulated, transported and packaged by qualified staff.
- The equipments used in the process (planter and thrasher) will be appropriately cleaned in the trial field. Corn grain will be harvested manually. We have provided a procedure for cleaning the equipment together with the documents of the B/ES/14/04 release.
- Transgenic and conventional corn plants will be crushed and buried in the soil.
- Alter the corn harvest, we will inspect visually the field and collect by hand any cop left to prevent the possible growth of seed remnants. In addition, we will control manually (by rogue or with herbicide) any volunteer that could appear in autumn or at the end of winter.
- The trial will be periodically checked in order to register any information about adverse effects towards the environment and / or food safety, which will be notified to the corresponding authority.
- In the end of the trial a report will be sent to the corresponding authority.

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:
01/01/1900 00:00:00
The Autonomous Community of Cataluña has issued a resolution expiration procedure.