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
Comparative analysis of morphological characteristics of genetically modified cucumber (Cucumis sativus L.) lines and non-GM line

Proposed period of release:
10/05/2014 to 31/08/2014

Name of the Institute(s) or Company(ies)
Institute of Plant Genetics, Polish Academy of Sciences;

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
cucumbercucurbitaceaecucumiscucumis sativushighly inbred line – Borszczagowski

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Introduced traits:
1. Sweet taste - GM cucumber lines with thaumatin II cDNA expression
2. Enhanced cold tolerance – GM cucumber lines with dehydrin 24 gene expression
3. Kanamycin resistance two groups of GM cucumber lines used for releasing: (a) with expression of
thaumatin II cDNA and (aa) with expression of dehydrin 24 gene.
Thaumatin II represents an unique class of extremely sweet-tasting plant proteins. As was confirmed by sensory
assessments of cucumber fruits, transgenic cucumber expression of the thaumatin II cDNA resulted in sweeter
taste in comparison with the control (non-GM fruits).
Dehydrin 24 – potential role in cold tolerance.
In vitro tests have shown that cucumber plants with expression of dehydrin 24 gene may be tolerant to low
The neomycin phosphotransferase II gene is commonly used for selection of transformed plant cells.
The gene codes for the aminoglycoside 3’-phosphotransferease II (NPTII) enzyme, which inactivates, by
phosphorylation, a range of aminoglycoside antibiotics such as: kanamycin, neomycin, geneticin, and
paromomycin. Kanamycin is commonly used in selective media.

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:
To the nuclear genome of cucumber, two genetic constructs were introduced: (1) with cDNA of thaumatin II from Thaumatococcus daniellii under the control of CaMV35S promoter and nptII gene from transposon T7, driven by NOS promoter (pNOS) (Szwacka et al., 2002, Acta Physiologiae Plantarum 24 (2): 173-185) and
(2) with Dhn24 gene from Solanum sogarandinum under the control of glucosyltransferase gene promoter and nptII gene under pNOS (Rorat T., Patent No 360191). Intended functions of introduced genes: (1) thaumatin II cDNA - introduction of sweet-taste phenotype, (2) Dhn24 –introduction of tolerance to low temperatures, (3) nptII – introduction of kanamycin-resistance (used for selection of transformed cells).

6. Brief description of the method used for the genetic modification:
For genetic modification of cucumber (Cucumis sativus L.) the agroinfection and in vitro culture methods were used (Szwacka et al., Genet. Pol. 37A, 1996, 126-129). Leaf microexplants (2-3 mm2) were inoculated with Agrobacterium tumefaciens culture. After infection and co-cultivation with agrobacteria, leaf explants were transferred onto selective medium supplemented with kanamycin (two-step selection of transformants: 100 and 150 μg/dcm3 ). The next step of modification procedure was regeneration of putative transformed plants from
selected kanamycin-resistant callus. The presence of foreign genes in nuclear genome of cucumber was carried out by Southern analysis.

Experimental Release

1. Purpose of the release:
The aim of the open field experiment is morphological analysis of genetically modified (GM) cucumber lines, carried out in comparison to the parental line (non-GM control). The morphological characteristics of fruits, leaves and flowers of cucumber plants will be compared.

2. Geographical location of the site:
Country – Poland
Voivodeship- wielkopolskie
City – Poznań

3. Size of the site (m2):
288 m2 with buffer zone and 160 m2 without buffer zone

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:
Earlier releasing of GMP (genetically modified cucumber plants with expression of
thaumatin II cDNA) – decision numbers: 5/2001,1/2002 and 3/2003. They confirmed no
adverse effects of GM plants on human health and environment (non-target organisms). Place of releasing – Experimental Field of Department of Vegetable and Medicinal Plants, Warsaw University of Life Sciences, Warszawa –Wilanów, Poland.

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
New traits introduced into the cucumber should not confer selective dominance in natural environment. Transgenic plants are influenced by the environment in the same way as non-transgenic varieties. Studies in open field carried out earlier with GM cucumber lines expressing thaumatin II cDNA have shown that yielding of GM plants with thaumatin II gene expression is comparable with yielding of no-GM control. Moreover, it was noticed that some GM lines are more tolerant to the pathogenic fungus Peronospora cubensis that causes a major foliar disease in cucumbers. It is expected that these lines will require less pesticide application.

Brief description of any measures taken for the management of risks:
Safe release of GMOs into the environment will be carried out according to the following plan:
-establishment of spatial isolation of GM cucumber plants (a minimum of 1000 m) from the other cucumber crops
-establishment of a buffer zone around GMO field, with a width of 2 m, planted non-GM cucumbers
-labelling of the GMO field (placing of informative boards)
-GM plants will be cultivated on the area monitored for 24 hours
-monitoring of the GMO field twice weekly
-after plant material harvest, plant residues (young fruits, leaves, flowers, shoots and roots) will be utilized and GMO field will be monitored to the end of the next growing season

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
We expect to confirm the lack of adverse effects of the released GM cucumber plants on the environment and human health.

Final report

European Commission administrative information

Consent given by the Member State Competent Authority:
21/01/2014 00:00:00