General informationNotification NumberB/IT/02/10Member State to which the notification was sentItalyDate of acknowledgement from the Member State Competent Authority30/12/2002Title of the ProjectEvaluation of transgenic parthenocarping processing tomato (Lycopersicon esculentum) in open field cultivationProposed period of release:01/05/2003 to 31/08/2003Name of the Institute(s) or Company(ies)UNIVERSITA' ANCONA - Facoltà di agraria -Dipartimento di biotecnologie agrarie e ambientali, ;
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) the enzyme tryptophan-2-monooxygenase which confers parthenocarpic fruit development;
2) the enzyme neomycin acetyl-phosphotransferase II which confers resistance to the antibiotics neomycin and kanamycin;
Thus the GMP display the following traits:
1. parthenocarpic fruit development referred to as the ability to set and develop normal seedless fruits under unfavourable environmental condition (T°, RU, light) for pollination and/or fertilization which prevent the normal development of carpel to fruit in untransformed tomato;
2. kanamycin resistance, determined either as the ability of plants to grow on kanamycin containing medium or to detoxify the antibiotic after spraying it on leaves of young plants which do not show any chlorotic symptom.
The GMP do not show any other phenotypical difference compared to untransformed wild type tomato.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 pCarp-iaaM which includes two physically linked expression cassettes:
- a 5.5 Kb chimeric gene, conferring parthenocarpic fruit development, obtained by a transcriptional fusion of the DefH9 promoter from Anthirrinum majus which allows specify expression in the ovules during flower and fruit development. The DefH9 promoter drives the expression of the codin region of the iaaM gene from Pseudomonas syringae pv.savastonoi which codifies for the enzyme triptophan-2-monooxygenase involved in the synthesis of the IAA auxin. To reduce the expression of the chimeric gene at posttranscriptional level a, 53 nucleotides immediately upstream the AUG initiation codon of iaaM sequence were replaced with 87 nucleotides derived by the rolA intron sequence frm Agrobacterium rhizogenes. The terminator of the rolC gene from Agrobacterium rhizogenes provides the transcription terminator signal.
- 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 transformants.6. Brief description of the method used for the genetic modification:Genetic transformation was achieved by in vitro infection of leaf, cotyledon and hypocotyls explants with the disarmed strain GV3101 of Agrobacterium tumefaciens carrying th binary plasmid pCarp-Ri-iaaM which harbours the chimeric T-DNA. Transgenic plants were selected in kanamycin containing medium.Experimental Release1. Purpose of the release:To verify the agronomical and technological characteristics of transgenic parthenocarpic industrial tomatoes under open field cultivation condition.2. Geographical location of the site:Ancona, Italy3. Size of the site (m2):Actual experimental trial size: 108 m2
Whole experimental size: 203 m2Environmental Impact and Risk ManagementSummary of the potential environmental impact from the release of the GMPts:In the site where the trials will be carried out, potential gene transfer from GMP is only possible to other tomatoes. No significant impact on the environment is expected.Brief description of any measures taken for the management of risks:
No uncontrollable spreading of GMP genetic material is anticipated.
- Minimal distance from other tomatoes plants will be at least 1 Km;
- Field will be set in restricted and controlled area;
- All fruits will be collected, boiled to denaturate tissue and seeds;
- At completation of the trial, the plants will be destroyed by using systemic herbicide active on tomato. The destroyed plants and the denaturated fruits will be incorporated into the soil, the area will be scouted to eliminate eventual volunteer tomatoes.Final report-European Commission administrative informationConsent given by the Member State Competent Authority:Yes20/03/2003 00:00:00Remarks: