Back

Notification report


General information

Notification Number
B/RO/10/05

Member State to which the notification was sent
Romania

Date of acknowledgement from the Member State Competent Authority
19/04/2010

Title of the Project
Notification according to Directive 2001/18/EC, Part B, for the deliberate release of of genetically modified DAS-Ø15Ø7-1xMON-ØØ6Ø3-6 maize for the use in field trials in Romania.

Proposed period of release:
01/04/2010 to 31/12/2014

Name of the Institute(s) or Company(ies)
Pioneer Hi-Bred Seeds Agro SRL, DN 2, km. 19,7
OP Afumaţi, Cod 077010, Ilfov, România;;


3. Is the same GMPt release planned elsewhere in the Community?
Yes:
Spain;

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

If yes, notification number(s):
B/DE/06/180; B/DE/10/207; B/ES/06/19; B/ES/07/11; B/ES/07/12; B/ES/08/16; B/ES/08/22; B/ES/09/26; B/ES/10/31; B/ES/10/39; B/FR/06/12/03; B/NL/06/07; B/RO/07/12;

Genetically modified plant

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

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
The genetically modified 1507xNK603 maize varieties that will be tested in these field trials are resistant to certain Lepidopteran insects, such as the European corn borer (Ostrinia nubilalis) and tolerant to glufosinate-ammonium and glyphosate herbicides. This maize was derived through traditional breeding methods between progeny of the genetically modified 1507 maize, which is resistant to certain lepidopteran insects and tolerant to glufosinate herbicide, and NK603 maize (from Monsanto), which is tolerant to glyphosate herbicide.

Genetic modification

3. Type of genetic modification:
Insertion; Other;
Other
Combination obtained by conventional crossing between genetically modified 1507 maize and genetically modified NK603 maize. . No additional genetic modification is involved.

In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
The genes introduced in 1507xNK603 maize are the cry1F gene from Bacillus thuringiensis var. aizawai, providing resistance to certain Lepidopteran insects and the pat gene from Streptomyces viridochromogenes, introduced as a selectable marker, providing tolerance to glufosinate-ammonium herbicide, like in 1507 maize, and the cp4epsps gene from Agrobacterium sp. strain CP4 that confers tolerance to glyphosate herbicide, like in NK603 maize, along with their regulatory components necessary for expression in maize plant:
- the cry1F gene, the UBIZM1(2) promoter, the ORF25PolyA terminator.
- the pat gene, the CaMV35S promoter, the CaMV35S terminator.
- the cp4epsps gene, the P-ract1/ract1 intron + ctp2, the NOS 3’ terminator.
- the cp4epsps gene, the CaMV e35S promoter + Zmhsp70 + ctp2, the NOS 3’ terminator.

Summary of the inserted DNA fragment inherited from 1507:
ubiZM1 PRO: 1.98 kb. The ubiquitin promoter (plus 5’ untranslated region) from Zea mays.
cry1F: 1.82 kb. A synthetic version of truncated cry1F from B. thuringiensis subsp. aizawai (plant optimized).
ORF25 TERM: 0.72 kb. A terminator from Agrobacterium tumefaciens pTi15955.
35S PRO: 0.55 kb. 35S promoter from Cauliflower Mosaic Virus.
pat: 0.55 kb. The synthetic glufosinate-ammonium tolerance gene (plant optimized), based on a phosphinothricin acetyltransferase gene sequence from Streptomyces viridochromogenes.
35S TERM: 0.20 kb. 35S terminator from Cauliflower Mosaic Virus.

Summary of the inserted DNA fragment inherited from NK603:

Genetic Element: Size (kb). Function.

- P-Ract1/ I-Ract1: 1.4kb. 5’ region of the rice actin gene containing the promoter, transcription start site and first intron.
- TS-CTP2: 0.2kb. DNA sequence coding for the N-teminal chloroplast transit peptide.
- CS-cp4 epsps: 1.4kb. DNA sequence coding for the native CP4 EPSPS protein.
- T-nos: 0.3kb. 3' transcript termination sequence of the nopaline synthase (nos) coding sequence from Agrobacterium tumefaciens which terminates transcription and directs polyadenylation.
- P-e35S: 0.6kb. Promoter and leader for the cauliflower mosaic virus (CaMV) 35S RNA (Odell et al., 1985) containing the duplicated enhancer region.
- I-Hsp70: 0.8kb. Intron from the maize heat-shock protein 70 gene.
- TS-CTP2: 0.2kb. DNA sequence coding for the N-teminal chloroplast transit peptide.
- CS-cp4 epsps l214p: 1.4kb. DNA sequence coding for the CP4 EPSPS L214P protein.
- T-nos: 0.3kb. 3' transcript termination sequence of the nopaline synthase (nos) coding sequence from Agrobacterium tumefaciens which terminates transcription and directs polyadenylation.
cp4 epsps gene cassette (1)
- P-ract1/ract1 intron
Source: Oryza sativa Intended function: 5’ region of the rice actin 1 gene containing the promoter, transcription start site and first intron. (1.4 kb)

- ctp 2
Source: Arabidopsis thaliana Intended function: DNA sequence for chloroplast transit peptide, isolated from Arabidopsis thaliana EPSPS, present to direct the CP4 EPSPS protein to the chloroplast, the site of aromatic amino acid synthesis. (0.2 kb)

- cp4 epsps
Source: Agrobacterium sp. strain CP4 Intended function: The DNA sequence for CP4 EPSPS, isolated from Agrobacterium sp. strain CP4, which imparts tolerance to glyphosate. (1.4 kb)

- NOS 3’
Source: Agrobacterium tumefaciens Intended function: A 3’ nontranslated region of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA which ends transcription and directs polyadenylation of the mRNA. (0.3 kb) cp4 epsps gene

second cassette

- e35S
Source: Cauliflower mosaic virus Intended function: The cauliflower mosaic virus (CaMV) promoter with the duplicated enhancer region. (0.6 kb)

- Zmhsp70
Source: Zea mays L. Intended function: Intron from the corn hsp70 gene (heat-shock protein) present to stabilize the level of gene transcription. (0.8 kb)

- ctp 2
Source: Arabidopsis thaliana Intended function: DNA sequence for chloroplast transit peptide, isolated from Arabidopsis thaliana EPSPS, present to direct the CP4 EPSPS protein to the chloroplast, the site of aromatic amino acid synthesis. (0.2 kb)

- cp4 epsps
Source: Agrobacterium sp. strain CP4 Intended function: The DNA sequence for CP4 EPSPS, isolated from Agrobacterium sp. strain CP4, which imparts tolerance to glyphosate. (1.4 kb)

- NOS 3’
Source: Agrobacterium tumefaciensIntended function: A 3’ nontranslated region of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA which ends transcription and directs polyadenylation of the mRNA. (0.3 kb)


6. Brief description of the method used for the genetic modification:
No genetic modification was made on the 1507xNK603 maize varieties directly: they were derived through traditional breeding methods between progeny of the genetically modified 1507 maize and NK603 maize.
1507 maize was genetically modified using the particle acceleration method. NK603 maize was genetically modified by Monsanto Company using particle acceleration method; the genetic modification was licensed to Pioneer for use in its own germplasm.


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 purpose of the trialing program is to evaluate the behavior in the environement and agronomic performances of 1507xNK603 maize varieties (inbreds and/or hybrids), including collection of data necessary for the registration of 1507xNK603 maize varieties on the National list of Romania.

2. Geographical location of the site:
The release is planned at the following location:
Dalga – Calarasi county;
Mircea Voda – Braila county;
Ramnicu Sarat – Buzau county;
Tecuci – Galati county;
Troianu –Teleorman county;


3. Size of the site (m2):
Dalga – Calarasi county-10000 sq.m
Mircea Voda – Braila county-10000 sq.m
Ramnicu Sarat – Buzau county -10000 sq.m
Tecuci – Galati county-10000 sq.m
Troianu –Teleorman county -10000 sq.m


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:
Field trials have already been conducted at several locations across maize growing regions in North America, South America and Europe and no environmental problems were reported for these trials. The transgenic plants behaved as non-genetically modified maize plants except for showing resistance to certain Lepidopteran insects such as Ostrinia nubilalis and showing tolerance when glufosinate and glyphosate herbicides have been applied, traits due to the genetic modifications.
1507xNK603 maize is no more regulated in the USA since 2001, in Canada since 2003, in Japan since 2005. It is widely cultivated and no adverse effects have been reported to our knowledge.


Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
No risks to human and animal health or the environment from the deliberate release of genetically modified 1507xNK603 maize resistant to certain Lepidopteran insects and tolerant to glyphosate and glufosinate herbicides are expected, as per information contained in the environmental risk assessment included in the notification:

• Likelihood of the genetically modified higher plant (GMHP) becoming more persistent than the recipient or parental plants in agricultural habitats or more invasive in natural habitats. There is negligible likelihood for 1507xNK603 maize to become environmentally persistent or invasive giving rise to any weediness. Maize does not possess any traits for weediness and the expression of Cry1F, PAT and CP4 EPSPS proteins in 1507xNK603 maize does not give rise to traits for weediness.
Characteristics for weediness have been generally described as 1) the ability for weed seed to germinate in many different environments; 2) discontinuous germination and great longevity of seed; 3) rapid growth through vegetative phase to flowering; 4) continuous seed production for as long as growing conditions permit; 5) self-compatibility but partially autogamous and apomictic; 6) ability to be cross-pollinated by unspecialized visitors or wind pollinated; 7) high seed output in favourable environments and some seed production in a wide range of environments; 8) adaptation for short and long distance dispersal; 9) vegetative production or regeneration from fragments and brittleness (hard to remove from the ground); and 10) ability to compete interspecifically by special means.
Maize does not exhibit the above weedy tendencies and is therefore non-invasive in natural ecosystems. Some Zea species are successful wild plants in Central America but they have no pronounced weedy tendencies. Maize has been domesticated to the extent that the seeds cannot be separated from the cob and disseminated without human intervention. Maize plants are annuals that generally will not survive in Europe from one growing season to the next because of poor dormancy and sensitivity to low temperature. Despite its non-dormant nature, maize seed can occasionally persist from one growing season to the next. When the temperature and moisture are adequate, the seed will germinate. This will happen under favorable climatic conditions, which is not the case in Northern latitudes where winters are very hard. In any case, these volunteers are easily identified and controlled through manual or chemical means.In case of unintended release of 1507xNK603 maize, current agronomic measures taken to control commercially available maize can be applied, such as use of herbicides (with the exception of glufosinate-ammonium and glyphosate herbicides), and manual or mechanical removal and incorporation in the soil.

• Any selective advantage or disadvantage conferred to the GMHP . As intended and when cultivated, expression of Cry1F, PAT and CP4 EPSPS proteins in 1507xNK603 maize confer specific advantages in agricultural environments: resistance to certain lepidopteran insect pests, such as the European corn borer and Sesamia sp., and tolerance to glyphosate and glufosinate-ammonium herbicides. Tolerance to glufosinate-ammonium is only used as a selectable marker for the screening process.
However, maize is highly domesticated, to the extent that it cannot become established as a feral species outside the agricultural environment due to its poor survival characteristics within European conditions. The specific advantages present in 1507xNK603 maize do not confer any selective advantage to the plants in the natural environment, i.e. outside the agricultural environment. Insect attack is one of the multiple biotic and abiotic factors that prevent growth of maize outside the agricultural environment. Therefore, expression of Cry1F protein conferring resistance to certain lepidopteran insect pests cannot be considered a selective advantage outside the agricultural environment.
Furthermore, expression of PAT and CP4 EPSPS proteins in 1507xNK603 maize does not confer a selective advantage outside the agricultural environment.

• Potential for gene transfer to the same or other sexually compatible plant species under conditions of planting the GMHP and any selective advantage or disadvantage conferred to those plant species. Spontaneously, there are no sexually compatible wild or weedy relatives of Zea mays in the European Union, which eliminates any potential for gene transfer to such species. Potential for gene transfer is therefore limited to other maize grown in culture. However, this potential will be considerably reduced by the trial conditions, for which a 200-meter isolation distance will be maintained between 1507xNK603 maize and any other non-experimental maize crop. In addition, the trial site will be surrounded by 4 border rows of conventional maize of a relative similar maturity that will be also destroyed at the end of the release.
As discussed in Point 2., above, the genetic modifications in 1507xNK603 maize do not introduce any selective advantages to maize plants outside the heavily managed agricultural environments.

• Potential immediate and/or delayed environmental impact resulting from direct and indirect interactions of the GMHP and target organisms, such as predators, parasitoids, and pathogens (if applicable). The genetic modifications in 1507xNK603 maize provide growers with a highly effective and environmentally beneficial tool to control certain lepidopteran insect pests, such as the European corn borer (Ostrinia nubilalis) and Sesamia spp. However, the maize efficacy would be reduced if the target insect pests develop resistance to Cry1F protein as expressed in 1507xNK603 maize during cultivation. The occurrence of such a phenomena, the target insect resistance, is negligible in the case of the trials in view of the restricted area of the release. The risk posed by the development of insect resistance is thus negligible. But at the time of commercial cultivation, a detailed proposal for insect resistance management (IRM) will be developed in the context of product stewardship and applied when 1507xNK603 is placed on the EU market. For example, implementation of refuges (zone cultivated with maize non- resistant to lepidopteran insects) will be set to ensure that rare insects that could become resistant to Cry1F toxin, and survive, can mate with sensitive insects. In this way, their progeny will be kept sensitive and will be controlled by future Bt maize crop.
No other potential immediate and/or delayed environmental impact resulting from direct and indirect interactions of 1507xNK603 maize and target organisms in the receiving environment are expected to arise from the proposed deliberate release of 1507xNK603 maize.

• Possible immediate and/or delayed environmental impact resulting from direct and indirect interactions of the GMHP with non-target organisms, (also taking into account organisms which interact with target organisms), including impact on population levels of competitors, herbivores, symbionts (where applicable), parasites and pathogens. The deliberate release of 1507xNK603 maize will result in negligible immediate and/or delayed environmental impact resulting from direct and indirect interactions of 1507xNK603 maize with non-target organisms, also taking into account organisms which interact with target organisms.
This conclusion is based on the specificity of the biological and biochemical activities of the Cry1F, PAT and CP4EPSPS proteins expressed in 1507xNK603 maize and the results obtained from ecotoxicity studies showing no toxic effects on a range of non-target organisms and beneficial insects, such as green lacewing larvae (Chrysoperla carnea), the ladybird beetle (Hippodamia convergens), the beneficial parasitic Hymenoptera Nasonia vitripennis, honey bee larvae (Apis mellifera), earthworms (Eisenia foetida), collembola (Folsomia candida), the cladoceran aquatic invertebrate Daphnia magna, and the non-target Lepidoptera Monarch butterfly, together with the absence of significant exposure of other non-target organisms, including non-target lepidopterans, to the Cry1F, PAT and CP4EPSPS proteins in the proposed release of 1507xNK603 maize for field trials.

• Possible immediate and/or delayed effects on human health resulting from potential direct and indirect interactions of the GMHP and persons working with, coming into contact with or in the vicinity of the GMHP release(s). Maize is not considered to have harmful effects on human health. In addition, maize has a long history of safe use in human food and animal feed. The genetic modifications in 1507xNK603 maize do not introduce any new compounds known to cause, or expected to cause, any potential immediate and/or delayed effects on human health resulting from possible direct and indirect interactions of the 1507xNK603 maize and persons working with, coming into contact with or in the vicinity of the release of 1507xNK603 maize.
The safety of the proteins Cry1F, PAT and CP4 EPSPS has been confirmed and 1507xNK603 maize does not pose any significant toxic or allergenic risk to humans. The placing on the market of 1507xNK603 maize for food and feed uses, import and processing in the European Union has been authorized by Commission decision 2007/703/EC.
Moreover, 1507xNK603 maize subject of the release will be destroyed and will not enter the food chain.

• Possible immediate and/or delayed effects on animal health and consequences for the feed/food chain resulting from consumption of the GMO and any products derived from it, if it is intended to be used as animal feed. 1507xNK603 maize has been obtained from traditional breeding methods between progeny of genetically modified 1507 and NK603 maize and no new compounds known to cause, or expected to cause, any possible immediate and/or delayed effects on animal health were introduced.
As previously mentioned, the placing on the market of 1507xNK603 maize for food and feed uses, import and processing in the European Union has been authorized by Commission decision 2007/703/EC.
Moreover, the 1507xNK603 maize plants subject of the current release are not intended for use in animal feed stuffs. The grain from the trials will be destroyed and will not enter the feed chain.

• Possible immediate and/or delayed effects on biogeochemical processes resulting from potential direct and indirect interactions of the GMO and target and non-target organisms in the vicinity of the GMO release(s). There is negligible likelihood that expression of Cry1F and PAT proteins in 1507xNK603 maize cause any possible immediate and/or delayed effects on biogeochemical processes resulting from potential direct and indirect interactions of the genetically modified maize and target or non-target organisms in the vicinity of the proposed release of 1507xNK603 maize, as discussed in Points 4 and 5 above.

• Possible immediate and/or delayed, direct and indirect environmental impacts of the specific cultivation, management and harvesting techniques used for the GMHP where these are different from those used for non-GMHPs. The specific cultivation, management and harvesting techniques used for the 1507xNK603 maize are identical to those used for non-genetically modified maize, with the exception of the herbicide regime (as glyphosate can be used for the weeding of the maize crop), containment measures (isolation distance and border rows) and destruction of the plant material (not used for food or feed). They are not expected to cause any possible immediate and/or delayed, direct or indirect environmental impacts.
For the needs of the studies, some plant tissue samples may be collected, manually or using a harvesting machine. When a few seeds need to be collected for analysis, this will be done by sampling the whole ear and unused seeds will be destroyed by incorporation into the soil. At the end of the release, all remaining plants matter that has not been harvested for analyses will be destroyed by chopping and incorporating into the soil by a deep ploughing. The border rows planted with conventional maize surrounding the trial site will be also destroyed at the end of the release by chopping and incorporating into the soil. No plant or plant product coming from the trials will enter the food or feed chains.
After the release, the plot will be visited regularly during the following year in order to ensure removal of maize volunteers, if any. Although volunteer maize cannot generally survive hard winter, maize volunteers, if any, will be monitored in order to ensure their destruction prior to flowering.


Brief description of any measures taken for the management of risks:
The pollen shed from the genetically modified plants will be controlled by maintaining a 200 meter isolation distance with any other commercial maize crop. As it is done for any trial with conventional maize, the trial site will be surrounded by agronomic border rows of conventional maize of a similar maturity that will be also destroyed at the end of the release.
Grains are fixed on a cob and enclosed in many husks that protect the seeds from outside contact. Thus seed dispersal of individual kernels is not likely to occur. When a few seeds need to be collected for analysis, this will be done by sampling the whole ear and unused seeds will be destroyed.
If needed, the trial could be stopped by mechanical destruction or application of a herbicide other than glyphosate and glufosinate-ammonium, and incorporation into the soil by a deep ploughing.
At the end of the release, all remaining plant matter that has not been harvested for analyses will be destroyed by chopping and incorporating into the soil by a deep ploughing. No plant or plant product coming from the trials will enter the food or feed chains.
After the release, the plot will be visited regularly during the following two years in order to ensure removal of maize volunteers, if any. Although volunteer maize cannot generally survive hard winter, maize volunteers, if any, will be monitored in order to ensure their destruction prior to flowering.
No commercial maize crop will be planted on the same plot the following two years.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Not applicable.
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:
Yes
03/06/2010 00:00:00
Remarks: