Ministry of the Environment Environmental Risks Department The Decision came into force on 121 April 2009. Prague, April 3, 2009 To Reference Number: 93849/ENV/08 D e c i s i o n The Ministry of the Environment of the Czech Republic as the administrative body competent according to § 5 of the Act No.78/2004 Coll., on the use of genetically modified organisms and genetic products as amended by the Act No. 346/2005 Coll. (hereinafter the „Act“) and in § 10 of the Act No. 500/2004 Coll., the Administration Code, as later amended, has decided on the basis of a request of the company BASF spol. s r.o. located in Šafránkova 3, 155 00 Praha 5, for granting consent for the deliberate release into the environment in the Czech Republic of genetically modified (GM) potatoes with modified starch composition, transformation event EH92-527-1 (Amflora) pursuant to § 5 par. 8 of the Act: the company BASF spol. s r.o. Šafránkova 3, 155 00 Praha 5 i s g r a n t e d c o n s e n t for the deliberate release of genetically modified potatoes with modified starch composition, transformation event EH92-527-1 (Amflora) into the environment in the Czech Republic. Requirements of the consent according to § 18 par. 6 of the Act: Authorised person 1 Name: BASF spol. s r.o., Šafránkova 3, 155 00 Praha 5 Identification Number (I ČO): 41195469 Specification of the genetically modified organism The genetically modified potato clone EH92-527-1, of commercial name Amflora derived from the clone P107 with inserted fragment of a gene gbss starch synthase bound on the starch grain in antisense orientation, which causes in the interaction with the promotor of gbss gene the decrease of amylase content and the increase of the amount of amylopectin in potato tuber starch. Therefore its starch differs from the starch produced by recipient variety only with regard to amylose/amylopectin content. The selective gene for neomycine phosphotransferase II ( nptII or APH(3´)II) from a mobile element (transposon) Tn5 of Escherichia coli inducing the resistance to the antibiotics kanamycin and neomycin during the selection of transformed plant tissue. The transformation was made in a standard way by using the construct Agrobacterium tumefaciens in a binary vector system. The genetically modified potato EH92-527-1 (Amflora) differs from conventional potato varieties only with regard to starch quality, and the presence of nptII gene. Agronomic characteristics are within the range corresponding to conventional potato varieties. No differences in interactions with non-target organisms are expected. Specification of the genetic modification To transform a potato the strain Agrobacterium tumefaciens LBA4404 with the content of Ti- plasmid pAL4404 was used. The inserted material (insert) is composed of the starch synthase promotor sequence bound with potato granule (990 bp, gbss ) and polyadenyl sequence of nopalin synthase gene (253 bp). The gbss gene segment in the inverted orientation to the promotor was inserted between these two DNA segments. The selective gene nptII (neomycin phosphotransferase II) with the nos (nopalin synthase from Agrobacterium tumefaciens) promotor and the polyadenyl sequence on the line sides were inserted. The insertion of a fragment of starch synthase gene ( gbss) bound with the starch granule (in antisense orientation to the fragment gbss potato promotor causes the reduction of amylase content in the potato tuber starch. It evokes the increase of amylopectin content in potatoes. The selective gene for neomycine phosphotransferase II induces the resistance to the antibiotic kanamycin during the selection of transformed plant tissue. Risk assessment results In respect of the manner of propagation, absence of wild compatible varieties in the Central Europe, and no-possibility of running wild, a potato has been considered as a relatively safe plant variety the spreading of which can be kept controlled. In this case the risk for its introduction into the environment lies more likely in the presence of active selective genes. Nevertheless, the gene nptII has been approved generally as safe for GMOs and food prepared from them (see the statement of the European Food Safety Authority – EFSA - of 13 th April 2007. 2 The current potato varieties are very demanding with regard to the agrotechnical conditions. There is no case of long-term survival of the plants in open air. In cold and low temperature conditions the potato plants are immediately contested by many fungal or viral diseases. Both the potato leaves and tubers serving for vegetative propagation are susceptible to frost damage. Due to an absolute absence of the blooming or defoliation of buds, the generative reproduction of the most of varieties is impossible. In many cases the varieties produce a small amount of pollen with the low fertility, often totally sterile. The plant is self-pollinating, and although it is pollinated almost entirely by insect, the flowers are not visited by main pollinators (bees). The creating of fruitages (berries) through the self-pollinating is not very common, and the berries after the earing early defoliate before the grains get matured or in consequence of the harvesting. It causes a significant decrease of the possibility that seeds could survive in soil. The seeds eventually survived loose the ability for germinating when ploughing under the soil into the depths of 0.5 cm. Potato seeds are not spread by birds. Relative species do not occur by nature in the Czech Republic with the exception of S. nigrum and S. dulcamara, with which a potato is not sexually compatible. An occurrence of potato plants run wild has not been observed as well. Not any difference has been observed concerning the survival and persistence in agricultural locations or the agressivity of penetration into the natural environment when compared to conventional potato varieties. A selective advantage or disadvantage does not exist in nature not even in consequence of the decrease of amylase content and the insert of nptII gene for the resistance of plants to the antibiotic kanamycin and other aminoglycoside antibiotics because of none-existence of a selective pressure which could give advantage to the virus carrier. The system can be used only for the selections in vitro conditions. Other selection advantage does not arise of this gene manipulation for potato plants. The both antibiotics have limited or none clinical importance because of the toxicity thereof. The resistance to the both antibiotics occurs in nature much frequently, and the genes of resistance are present in natural populations of microorganisms. The safety of APH(3´)II with regard to human and animal health has been evaluated in the publication series, and it has been also confirmed by quite a few standpoints of the Scientific Committee on Plants and the European Food Safety Authority (EFSA) (FDA, 1998; VIB, 2001; EFSA, 2004). The EFSA Scientific Committee has carried out a detailed risk assessment, and stated that there is no harmful effect caused by genetically modified event EH92-527-1 (EFSA, 2006a; EFSA, 2006b). Tubers are the useful part of the plant. The rigorous control of the content of harmful substances in tubers is carried out in the used varieties, and the limit values for the variety authorisation are determined. The genetic modification above mentioned is not intended to influence the content of harmful substances, and so any changes in the amount thereof are not awaited. Moreover, the tested genetically modified line is not intended for the use as food or feed, and therefore the input of inserted genes into food chains is decreasing to minimum. Agronomic characteristics represent a range of common potato varieties. No changes in the interactions with non-target organisms are awaited. It is expected that possible interactions of genetically modified potato clones with the non-living environmental elements will be comparable with common starch potatoes. The material from field experiments for EH92- 527-1 (Amflora) has been analysed, and it is proved that a starch modification had been stabile during several tuber generations. No other differences between EH92-527-1 (Amflora) clone and common used starch potatoes in the agronomic characteristics in relation to the introduction of potatoes into the environment. Because no changes in the agronomic characteristics are introduced, the lines of genetically modified potatoes show the same properties relative to the environment during cultivation as the comparable commonly used starch potatoes. 3 The request for the marketing of EH92-527-1 (Amflora) clone including the cultivation, has been in the EU submitted under both the Directive 2001/18/EC (request No. C/SE/96/3501) and the Regulation 1829/2003 (request EFSA-GMO-UK-2005-14). In both cases the European Food Safety Authority (EFSA) stated its positive opinion on the supposed use. Field trials with Amflora potatoes under given conditions will have the same impact on the environment as the cultivation of any other potatoes. Conditions for the use Genetically modified organisms above mentioned shall be used only in the way described in the request BASF spol. s r.o. located in Šafránkova 3, 155 00 Praha 5, Ref. No. MŽP 93849/ENV/08) submitted to the Ministry of the Environment on December 23, 2008, and supplemented by the submission delivered to MoE on March 10, 2009 (this request relates to the request of the company PROAGRO Radešínská Svratka, a.s., located in Radešínská Svratka 134, 592 33 Radešínská Svratka, Ref. No. MŽP 94262/ENV/08, and only when keeping all given conditions as follows: - The field trials shall not been carried out on the plot in location Svrate čka (a certified planting region). The data on this location are given only in the Annex to the request “Havarijní plán” (Emergency Plan) under point b).