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
Testing of NK603 (MON-ØØ6Ø3-6) maize. Request of the revision of the consent for deliberate releasing into environment for testing of genetically modified plant of maize (Zea mays -NK603), no 9 din 26 April 2006 – by the Romanian Ministry of Environment and Water Management.

Proposed period of release:
15/04/2007 to 01/12/2008

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

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

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

Genetically modified plant

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

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
NK603 Roundup Ready maize was produced by the introduction of a glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from Agrobacterium sp. strain CP4 (CP4 EPSPS) into the maize genome. The expression of glyphosate-tolerant CP4 EPSPS proteins in crop plants imparts tolerance to glyphosate (N-phosphonomethyl-glycine), the active ingredient in the non-selective, foliar-applied, broad-spectrum, post-emergent herbicide Roundup®2. The use of maize plants containing the Roundup Ready gene enables the farmer to utilise Roundup herbicide for effective control of weeds during the growing season and to take advantage of the favourable environmental and safety characteristics of Roundup herbicide.

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:
NK603 Roundup Ready maize was generated using a particle acceleration transformation system and a gel-isolated MluI fragment, PV ZMGT32L, containing a 5-enolpyruvyl¬shikimate-3-phosphate synthase (EPSPS) gene from Agro¬bacterium sp. strain CP4 (CP4 EPSPS). The cp4 epsps gene encodes a tolerant form of EPSPS, which confers glyphosate (Roundup) tolerance to the plant.
The plant expression plasmid vector, PV-ZMGT32, was developed by Monsanto Company, St. Louis, Missouri and contains two adjacent plant gene expression cassettes each containing a single copy of the cp4 epsps gene. The vector also contains an nptII selectable marker gene, which allows selection of bacteria containing the plasmid, and an origin of replication (ori) necessary for replicating the plasmid in Escherichia coli.
The agarose gel-isolated MluI restriction fragment of plasmid vector, PV ZMGT32L, which was utilized for transformation of NK603 Roundup Ready maize, contains only the CP4 EPSPS plant gene expression cassettes and does not contain the nptII selectable marker gene or origin of replication.
All genetic constituents within plasmid vector PV-ZMGT32 are completely known including the gene of interest, cp4 epsps, and its function. The agarose gel-isolated MluI restriction fragment of the plasmid vector, PV ZMGT32L, utilized for transformation of NK603 Roundup Ready maize, contains only the CP4 EPSPS plant gene expression cassettes and does not contain the nptII selectable marker gene or origin of replication.
In NK603 Roundup Ready maize, the e35S promoter utilised in the first plant gene expression cassette, as derived from cauliflower mosaic virus (CaMV), is well characterised. The safety of the e35S promoter is well established and does not impart a pathogenic response. The sequence for epsps utilised in both cassettes was isolated from Agrobacterium sp. strain CP4, while the NOS 3’ terminator is a 3’ non-translated region of the nopaline synthase gene derived from the Ti plasmid of Agrobacterium tumefaciens. There is no human or animal pathogenicity known from Agrobacterium species, nor is the cp4 epsps or NOS 3’ sequence a determinant of Agrobacterium plant pathogenesis.
The P-ract promoter and intron, Zmhsp70 intron and ctp2 inserted genetic elements are derived from common plant species including rice, maize and Arabidopsis.
In conclusion none of the inserted sequences are known to have any pathogenic or harmful characteristics.

6. Brief description of the method used for the genetic modification:
NK603 Roundup Ready maize was modified by incorporation of a restriction fragment of plasmid DNA, designated as PV-ZMGT32L, into the maize genome using a particle acceleration method.

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 this release is to collect data necessary for the registration of new NK603 maize hybrids on the National list of Romania.

2. Geographical location of the site:
Delibetrate release into the environment is planned for 2007 in the following locations:
• The official network of State Institute for Variety Testing and Registaration (ISTIS) where NK603 hybrids will be tested for collecting the necessary data for registration:
CTS Tecuci, Galaţi county, Lat.= 45.80993, Long.=27.40142
CTS Mircea Vodă, Brăila county, Lat.=45.10785, Long.=24.40990
CTS Dâlga, Călăraşi county, Lat.=44.42489, Long.=27.03711
CTS Râmnicu Sărat, Buzău county, Lat.=45.39225, Long.=27.06995
CTS Troianu, Teleorman county, Lat.=44.00437, Long.=25.02520

3. Size of the site (m2):
In 2007, the hybrids of genetically modified maize NK603 will be tested in each of the five testing locations in plots of 4 rows, 70 cm apart, containing maximum 100 plants (plot area 18-20 m2 accordingly to the plant populations) in 5 replications, summing an area of maximum 100 m2/hybrid/location. It is very probable that in the next years, genetically modified maize, subject of this application, would be planted in the same locations and on the same areas like in 2007.

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 were indistinguishable from the non-genetically modified maize plants except for showing tolerance when glyphosate herbicides have been applied, traits due to the genetic modifications.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
Potential environmental impact from the release or the placing on the market of GMOs (Annex II, D2 of Directive 2001/18/EC), if different from a similar release or placing on the market of the recipient or parental organism(s). Analysis of the characteristics of NK603 maize has shown that the likelihood of potential adverse effects on human health and the environment in the European Union, resulting from its import and use as any other maize, including the cultivation of NK603 maize varieties and the use of this maize in animal feed, is consistently negligible. Therefore, the overall environmental risk posed by the GMHP is also negligible, and strategies for risk management for NK603 maize would be the same as for traditional maize.
It is actually expected that the production of Roundup Ready maize will positively impact current agronomic practices in maize and provide benefits to farmers and the environment. The use of Roundup in maize enables the farmer to take advantage of the herbicide’s favourable environmental and safety properties. NK603 Roundup Ready maize benefits the farmer by providing (1) an additional broad-spectrum weed control option in maize, (2) a new herbicidal mode of action for in-season maize weed control, (3) increased flexibility to treat weeds on an “as needed” basis, (4) cost-effective weed control and (5) an excellent fit with reduced-tillage systems. In turn, a number of environmental benefits arise from the use of conservation tillage including improved soil quality, improved water infiltration, reductions in erosion and sedimentation of water resources, reduced runoff of nutrients and pesticides to surface water, improved wildlife habitat, increased carbon retention in the soil, reduced fuel use and encouragement of sustainable agricultural practices. Roundup herbicide has a very favourable environmental and health and safety profile. Glyphosate, the active ingredient in Roundup herbicide (1) has limited mobility as it binds rapidly and tightly to a wide variety of soils and sediments, (2) is non-persistent as it is readily metabolised, (3) has a low potential to move into surface or groundwater and (4) does not accumulate in and presents minimal risk to terrestrial and aquatic animals including birds, mammals, fish and invertebrates.
Potential environmental impact of the interaction between the GMHP and target organisms (if applicable), if different from that of the recipient or parental organism(s)NK603 maize is herbicide tolerant and, as such, has no target organisms with which to interact, either directly or indirectly.
Possible environmental impact resulting from potential interactions with non-target organisms, if different from that of the recipient or parental organism(s)
(a) Effects on biodiversity in the area of cultivation. Cultivated maize interacts with a range of organisms in the area of cultivation, including pathogens, micro-organisms, vertebrate wildlife and numerous soil dwelling and foliar dwelling invertebrates. As NK603 maize was shown to be equivalent to traditional maize, except for the introduced glyphosate-tolerance trait, its baseline interaction with these organisms is considered no different from traditional nontransgenic maize, except for the potential additional exposure of herbivorous pests of maize to the CP4 EPSPS proteins that are newly expressed in the plant. Through trophic transfer and decomposition processes, additional organisms in the field such as predators and prey of the pests of maize could theoretically be exposed to some very low levels of these proteins.
CP4 EPSPS proteins expressed in NK603 maize belong to the larger family of EPSPS enzymes, which are ubiquitous in bacteria, fungi, algae and plants. Therefore, there is no a priori reason to suspect that any EPSPS protein, including the CP4 EPSPS protein derived from the soil-borne bacterium Agrobacterium sp. strain CP4, would possess biological activity towards non-target organisms. Any non-target organisms interacting with the crop have co evolved in close interaction with a wide spectrum of green plants and microorganisms, and therefore have historically been exposed to members of this safe class of proteins. Numerous field trials have confirmed that the susceptibility of NK603 maize to insect pests or fungal diseases is no different from that of traditional maize varieties. These results are indicative of the fact that the interaction between this maize and non-target organisms is not different from traditional maize. In addition, no evidence has been brought forward by the many farmers who have grown NK603 maize since 2001 (or other Roundup Ready products that express CP4 EPSPS proteins since 1996) of any harmful or undesirable effects on non-target organisms.
In conclusion, no adverse effects on non-target organisms or indirect population effects on depending organisms are expected as a result from the import or use of this maize as any other maize, including the cultivation of NK603 varieties.
(b) Effects on biodiversity in other habitats. Like traditional maize, NK603 maize is a poor competitor and is non-invasive of natural environments. Therefore, the potential of this maize to cause adverse effects to wild plants in surrounding habitats, e.g. through competition for natural resources, is negligible. Maize can also interact with animals in habitats surrounding the crop. For instance, as maize is a good source of nutrition, interactions with vertebrate wildlife are well known. However, as NK603 maize is substantially equivalent to traditional maize, except for the introduced glyphosate-tolerance trait, its baseline interactions with these organisms will not be different from traditional maize, except for the potential additional exposure to the expressed CP4 EPSPS proteins. The CP4 EPSPS proteins are members of a well-known family of proteins that are ubiquitous in plants, algae, fungi and bacteria. There is no known toxicity associated with this family of proteins, which has been confirmed in toxicity tests and nutritional evaluations. The results of these tests indicate that NK603 maize or the expressed CP4 EPSPS proteins have negligible potential to cause any immediate or delayed adverse effects in vertebrates or invertebrates that are interacting with or even foraging on NK603 maize plants in the field. In the absence of potential direct adverse effects on pathogens, wild plants, invertebrates and vertebrates, no indirect population effects are to be expected in any other species that would predate on, interact with or otherwise depend on these organisms in the receiving environment.
It is concluded, therefore, that the likelihood of potential direct or indirect adverse effects in non-target organisms coming into contact with NK603 maize is negligible, and the risk posed by NK603 maize, therefore, is also negligible.

Brief description of any measures taken for the management of risks:
The shedding of pollen from the genetically modified plants will be controlled by maintaining a 200-meter isolation distance with any other non-experimental maize crop. In addition, the trial site will be surrounded by four border rows of conventional maize of a similar relative maturity that will be also destroyed at the end of the release.
Seed dispersal of individual kernels does not generally occur. They are fixed on a cob and enclosed in many husks that protect the seeds from outside contact. 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. In case of any emergency, the trial could be stopped by application of a non-selective herbicide, other than glufosinate and glyphosate, or by mechanical destruction and incorporation into the soil.
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. No plant or plant product coming from the trials will enter the food or feed chains.
Volunteer maize will be monitored during the following year in order to ensure their destruction, if needed.
No commercial maize will be cultivated in the trial site the following year.

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:
23/04/2007 00:00:00
It also includes the import permit no. 11/23.04.2007