General informationNotification NumberB/IT/04/03Member State to which the notification was sentItalyDate of acknowledgement from the Member State Competent Authority25/10/2004Title of the ProjectEVALUATION OF TRANSGENIC LEMON (Citrus limonL. Burm. f.) FOR RESISTANCE TO FUNGAL PATHOGENSProposed period of release:01/01/2005 to 31/12/2015Name of the Institute(s) or Company(ies)University of Catania, Facoltà di Agraria - Dipartimento di OrtoFloroArboricoltura e Tecnologie Agroalimentari;
3. Is the same GMPt release planned elsewhere in the Community?NoHas the same GMPt been notified elsewhere by the same notifier?NoGenetically modified plantComplete name of the recipient or parental plant(s)
2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:Two genes were inserted into the genomic DNA which encode for two proteins with different function:
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1) an endochitinase which degrades the cellular wall in superior fungi
2) the enzyme neomycin acetyl-phosphotransferase II which confers resistance to the antibiotics neomycin and kanamycin.
Thus, GMPs, display the following traits:
i) Due to the production of the endochitinase which has effect on a wide range of fungal pathogens and high antibiotic activity, the GMP turn out to be more resistant to the fungal infection. This trait will be evaluated by several tests with particular reference to Phoma tracheiphila.
ii) kanamycin resistance, determined either as the ability of plants to growth on a kanamycin containing medium or to detoxify the antibiotic after spraying it on leaves of young plant which do not show any chlorotic symptom.
The GMPs do not show any other phenotypical difference compared to untransformed wild type lemon.Genetic modification3. 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:The inserted DNA region corresponds to the T-DNA of Agrobacterium plasmid pBin19 which includes two physically linked expression cassettes:
- the chimeric gene CaMV35S-chit42-NOS formed by subunit 35 of cauliflower mosaic virus as promoter and NOS terminator from the nopaline synthetase gene of Agrobacterium tumefaciens, which regulate the expression of the codificant region of chit42 gene, isolated by micoparasite Trichoderma harzianum CECT 2413. The function of this fragment is expected to be the synthesis of endochitinase which demolishes cellular wall in fungi in order to prevent infection.
- a 1.5 Kb chimeric gene, conferring kanamycin resistance phenotype, obtained by the insertion of the neomycin phosphotransferase (NPTII) gene from Escherichia coli which codifies for the detoxifying enzyme, between the NOS promoter and the NOS terminator from the nopaline synthetase gene of Agrobacterium tumefaciens. It is the selectable marker employed to select for the transformants.6. Brief description of the method used for the genetic modification:Genetic transformation was achieved by in vitro infection of internodes explants with the disarmed strain EHA 105 of Agrobacterium tumefaciens carrying the binary plasmid pBin19 with the chimeric genes CAM35S-chit42-NOS and NOS-nptII-NOS. Transgenic plants were selected in kanamycin containing medium.7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:The GMP is not a forest tree species.Experimental Release1. Purpose of the release:The purpose of the release is to verify, under normal culture conditions, the agronomic characteristics of transgenic clones and the effect of chit42 gene on the tolerance to fungal pathogens. Another purpose is to analyse the effects on the environment, especially the genic flux to other sexual compatible plants and the possible effect on the non-target organism.2. Geographical location of the site:The release will be carried out in one location:
Azienda Agraria Sperimentale of Agraria Faculty – University of Catania, sited in Primosole (contrada), Catania.3. Size of the site (m2):Production field trial: 120 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:This GMP has never been released in environment.Environmental Impact and Risk ManagementSummary of the potential environmental impact from the release of the GMPts:The GMP does not feature higher dissemination possibility and, the only selective advantage is the higher resistance to fungal diseases. Thus, it is not deemed that such a genetic modification is likely to lead the transgenic clones to prevail over the non-transgenic ones.
A narrow use of pesticides, as a consequence of the resistance to fungal pathogens, determined by the exogenous gene, would entail a remarkable benefit for environment and for human health.Brief description of any measures taken for the management of risks:In order to prevent genetic flux, plants will be protected by metallic webs and anti-insect webs; trial field will be set in place not bordering on other citrus fields and it will be surrounded by monoembrionic citrus whose seeds will be analysed to find possible hybridation.
All the GM fruit will be collected and boiled to inactivate tissues and seeds. At completion of trial, plants will be eradicated and burned.Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:The release of this GMP is not deemed to have any impact on environment and on human health.Final report-European Commission administrative informationConsent given by the Member State Competent Authority:Not known