Date of publication: March 25, 2004
Source:
http://gmoinfo.jrc.it/gmp_browse_geninf.asp
Notification
report
General information
Notification Number:
B/IT/03/01
Member State:Italy
Date of Acknowledgement:10/12/2003
Title of the Project:
"Italian Lycopersycon (ITA.LYCO): Biologia avanzata e
innovazione di processo al servizio della qualità del pomodoro
da industria italiana"
Proposed period of release From:31/03/2004
To:31/08/2004
Name of the Institute(s) or Company(ies): Metapontum
Agrobios s.c.a r.l.;
3. Is the same GMPt release planned elsewhere in the
Community?
No
4 - Has the same GMPt been notified elsewhere by the same
notifier?
No
Genetically
modified plant
1. Complete name of the
recipient or parental plant(s)
|
Common Name |
Family Name |
Genus |
Species |
Subspecies |
Cultivar/breeding line |
| tomato
|
solanaceae
|
lycopersicon |
lycopersicon esculentum |
esculentum
|
Red Setter
codice varietà O 1270 |
2. Description of the traits and characteristics which have
been introduced or modified, including marker genes and previous
modifications:
From a genetic point of view, modifications brought to GMPt
consist in the insertion within the genomic DNA of two
transgenes:
- tLcy cDNA isolated from tomato and coding for the lycopene
cyclase enzyme. In the expression cassette this coding sequence
is under the control of 35SCaMV promoter and the NOS terminator.
Lycopene cyclase catalyses the cyclization of lycopene in
beta-carotene.
- NptII gene that codifies for the enzyme neomycin
phosphotransferase II which catalyses the phosphorilation of the
antibiotics neomycin and kanamycin.
As a consequence of the genetic modifications GMPt has got the
following traits:
- Overexpression of lycopene cyclase in tomato plants. This
modification allows the accumulation of beta-carotene instead of
lycopene in the tomato fruits. In the transgenic plants the ripe
fruit has a deep orange colour.
- Kanamycin resistance phenotype, determined as the ability of
plants or tissues to grow on kanamycin containing selective
media.
Genetic
modification
3. Type of genetic
modification:
Insertion;
4. In case of insertion of genetic material, give the source
and intended function of each constituent fragment of the region
to be inserted:
Inserted DNA region corresponds to the Agrobacterium plasmid
pJLcy T-DNA, which includes two physically linked expression
cassettes:
- 2780 bp chimeric cassette, which confers the high
beta-carotene phenotype, obtained by the transcriptional fusion
of the 35S promoter (850 bp) of the Cauliflower Mosaic Virus
(CaMV) which allows constitutive expression in transgenic
tissues, tLcy gene from tomato (1560 bp) coding for lycopene
cyclase enzyme, and NOS terminator (300 bp), from the nopaline
synthase gene of Agrobacterium tumefaciens, which carries the
termination signal for transcription.
- 1566 bp chimeric construct, which confers the kanamycin
resistance phenotype, obtained by the insertion of the neomycin
phosphotransferase Il gene (nptII of 792 bp) which codifies for
the detoxifying enzyme, between the NOS promoter (474 bp) and
the NOS terminator (300 bp) from the nopaline synthase gene of
Agrobacterium tumefaciens, which allow respectively the start
and the end of transcription in transgenic tissues. Up to date,
nptII is the most selective marker gene used in plant
transformation and kanamycin is the antibiotic more frequently
used for the selection of nptII-transgenic plants (Brasileiro
A:C:M and Aragao F:J:L:, 2001, J.Plant Biotechnology 3 (3):
113-221).
6. Brief description of the method used for the genetic
modification:
Plasmid pJLcy containing the tlcy-b sequence was transferred
to A. tumefaciens LBA4404 by electroporation. Tomato
(Lycopersicon lycopersicum L.Karsten ex Farw cv Red Setter)
cotyledons were transformed as described (Van Roekel, J.S.C., et
al. 1993. Plant Cell Rep. 12, 644-647.), except that the time of
co-cultivation was of 10 min and no feeder layer was used.
Putative transformants were selected on Kanamycin containing (50
μg/ml-1) substrate. Selected plants were advanced in greenhouse
and checked by Southern blot analysis. The resulting transgenic
plants were self-fertilised (D’Ambrosio C. et al. 2004 Plant
Science 166: 207-214). Four generations were necessary to get
the homozygous HC3B6 line.
7. If the recipient or parental plant is a forest tree
species, describe ways and extent of dissemination and specific
factors affecting dissemination:
The GMPt of this release is not a forest tree.
Experimental
Release
1. Purpose of the release:
The purpose of this release will be the evaluation of the
biological risk derived by the cultivation of the GMP in open
field in a strictly controlled experimental field. Concurrently,
GMP will be evaluated for some agronomical characteristics in
comparison to other two varieties: the REDSETTER, from which GMP
was derided, as well as an elite commercial variety. Biological
risk evaluation of the GMP release will be mainly concentrated
on the possible dissemination of transgenic pollen to the sides
of the experimental plot which will be covered with plants of
REDSETTER variety as test plants. Fruits produced by test tomato
plants will be collected in order to obtain seeds potentially
derived from fertilization of ovules with transgenic pollen.
Seeds will be planted in greenhouse and plants allowed to set
fruit. Plants will be inspected to look for orange coloured
fruits that demonstrates the pollen transfer.
2. Geographical location of the site:
The trial will be located at Metaponto (MT) in Southern
Italy, in a field of Pantanello Farm, which belong to Basilicata
Region Local Government (Regione Basilicata) 100 meters far the
"Metapontum Agrobios" Research Centre (S.S. Jonica 106, Km
448.2, Metaponto (MT), ITALY.
3. Size of the site (m2):
6400 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:
No previous release were carried out with this GM-plant.
Environmental
Impact and Risk Management
Summary of the potential
environmental impact from the release of the GMPts:
No factor unique to this field test has been identified that
would have an effect on any plant or animal species. In these
plants, there were no intentional changes influencing their
susceptibility to disease causing organisms. Moreover there are
no reasons to believe that the GMPt peculiarity (high content of
beta-carotene in ripe fruits) affects the dissemination and the
palatability of fruits and seeds to insects. In fact, natural
mutants having similar characteristics exist and none difference
has been detected. Cross pollination with wild species is not
possible because in this environment no wild type species
compatible with tomato are present. Crosses of the transgenic
pollen with tomato commercial varieties will be avoided since
the plot will be surrounded by maize and tomato barriers and is
fairly distant from other tomato fields. Moreover, previous
field test conducted from the notifier (Not. B/IT/00/09) on a
different GMPs demonstrated that there was no transfer of
exogenous DNA from the transgenic plants to the tomato barrier
plants (Grieco P.D. et al., 2001, Relazione finale su POM A15
"Scelte tecnologiche del sistema Agro-alimentare e qualità dei
prodotti" A1: 1-31).
Brief description of any measures taken for the management of
risks:
The field trial will take place on one small plot on
agricultural land in the Basilicata (ITALY). The site will
provide adequate physical security. Site monitoring of the field
trial and agronomic management practices creating controlled
environment are expected to provide the necessary degree of both
biological and physical containment. Pollen dispersal will be
avoided by maize and tomato barriers, which will surround the
entire field. After sample collection and fruit harvesting, the
remaining plants or parts will be removed, autoclaved and/or
burned and finally disposed off. Possible plant residues left on
the field will be harvested and burned on site. The test area
will be marked to monitor re-emergence of volunteer tomato
plants during the following seasons. The plots will not be
planted the following season but will be checked several times
to destroy any plant material.
Summary of foreseen field trial studies focused to gain new
data on environmental and human health impact from the release:
The GMP under discussion does not possess any form of
resistance or tolerance to pathogens or insects, so it does not
have any selective advantage respect to the control plant.
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