Summary Notification

General information

Notification Number
B/DK/12/01

Member State to which the notification was sent
Denmark

Date of acknowledgement from the Member State Competent Authority
08/03/2012

Title of the Project
Field trial og PAPhy07-Cisgenic barley with improved phytase activity.

Proposed period of release:
01/05/2012 to 30/09/2012

Name of the Institute(s) or Company(ies)
Aarhus University, Forskningscenter Flakkebjerg;

3. Is the same GMPt release planned elsewhere in the Community?
No

Has the same GMPt been notified elsewhere by the same notifier?
No

Genetically modified plant

Complete name of the recipient or parental plant(s)

Common Name	Family Name	Genus	Species	Subspecies	Cultivar/breeding line
barley	poaceae	hordeum	hordeum vulgare	vulgare	Golden Promise

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
The genetically modified barley line PAPhy07 has an increased activity of the HvPAPhy_a phytase enzyme due to the insertion of an extra HvPAPhy_a gene derived from Hordeum vulgare L. subsp. vulgare (variety Igri). The phytase activity has increased from 1350 FTU/kg flour to 3550 FTU/kg flour. There are no selection genes or vector-backbone sequences present in the GM barley line. The GM barley line has not previously been modified.

Genetic modification

3. Type of genetic modification:
Insertion; Deletion;

In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
The genetically modified barley line PAPhy07 contains one extra copy the HvPAPhy_a gene which codes for the HvPAPhy_a phytase protein derived from Hordeum vulgare L. subsp. vulgare (variety Igri). The phytase is synthesized in the seed during seed development and stored in the mature grain.
A full description of the genetic elements in PAPhy07, including the size and function is provided in Table 1.
Table 1. Summary of genetic elements inserted in the genetically
modified barley line
___________________________________________________________
Genetic Size Function
elements (bp)
___________________________________________________________

Left border 16 Derived from the 25 bp nucleotide
sequence that delimits the T-DNA
transfer into plant cells, originally
isolated from A. tumefaciens nopaline
nopaline vector pTiT37

Synthetic bp 14 Derived from the multiple
cloning site in pClean-185 used for
the transformation

HvPAPhy_a 5208 The fragment contains the coding
sequense of HvPAPPhy_a on 2266 bp, the
promotor flanking region of 2180 bp and the terminator flanking region of 762bp

Synthetic bp 22 Derived from the multiple cloning site
in p Clean-185 used for the trans- formation.

Right border 3 Derived from the 25 bp nucleotide
sequence that delimits the T-DNA
transfer into plant cells, originally
from A.tumenfaciens nopalin vector
pTiT37

In case of deletion of genetic material, give information on the function of the deleted sequences:
The integration of the HvPAPhy_a insert has caused a deletion of 22-bp at the insertion site. We have isolated 1056-bp and 734-bp on the RB and LB site of the insertion site, respectively and as judged from database searches these sequences do not appear to be functional.

6. Brief description of the method used for the genetic modification:
The genetically modified PAPhy07 was produced by Agrobacterium-mediated transformation of immature embryos.

7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:
Not applicable

Experimental Release

1. Purpose of the release:
The purpose of the release is to study the phytase activity in the grains of the genetically modified barley line PAPhy07 when grown under field conditions.

2. Geographical location of the site:
The release site is located on the experimental area of Research Centre Flakkebjerg near Slagelse, West Zealand.

3. Size of the site (m2):
50 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

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
We have developed and selected the genetically modified barley line PAPhy07, so that it is very close to the mother cultivar. PAPhy07 contains a single extra HvPAPhy_a phytase gene isolated from the barley itself and the line does not contain selection genes or sequences of the vector backbone and is therefore without antibiotic resistance genes. In addition, there is no coding or known functions in and around the region in which the insert is integrated so that the insert is not interrupted coding sequences, or well-known functions in the genome. The barley line is also phenotypically identical to the recipient.

• The risk for that the introduced trait in the genetically modified barley line PAPhy07 willgive the modified line a competitive advantage or disadvantage in natural environments is negligible. Unintended spreading of PAPhy07 in the non-agricultural environments is negligible. Barley is a cultivated crop with poor competitiveness outside agricultural areas. Insertion of an additional barley phytase gene in barley will only give rise to the production of proteins that are already present in the plants. This will not make the GM barley lines more persistent and invasive or provide selective advantages or disadvantages.

• The genetically modified barley line PAPhy07 may cause undesirable effects if it spreads to other barley fields by pollen or seeds. This can cause unintended mixing with non-GM seed lots and can have significant consequences, since seed lots with GM content not approved for cultivation in the EU are not allowed. The probability that seeds from hybrids or spilled seed from the GM barley line PAPhy07 could be mixed into other seed lots is, however, negligible. The outcrossing rate in barley is very low, and measures will be taken to avoid hybridization with other cultivated barley plants. Likewise, several measures will be taken to avoid the unintentional mixing of GM and non-GM seeds at sowing, harvesting and threshing and during transportation.

• It is highly unlikely that the genetically modified barley line PAPhy07 would be hazardous to non-target organisms. The ecological interactions with non-target organisms or the biochemical processes in soil are considered similar to the ones caused by conventional barley lines.

• Occupational health aspects of handling the genetically modified barley line PAPhy07 would not be different from conventional barley. There is nothing to suggest that the increased level of HvPAPhy_a phytase protein in the grains has negative consequences for human and animal health. HvPAPhy_a phytase enzyme is also present in non-GM barley plants. The increase in HvPAPhy_a phytase activity in the grains of the GM barley line is about the same as the corresponding phytase activity in grains of rye and some lines of wheat. The phytase activity in rye and wheat has no known harmful effects on humans and animals.

• The environmental impact of the cultivation, management and harvesting techniques applied in the planned trials is considered no different from the cultivation of any barley line.

Brief description of any measures taken for the management of risks:
The only potential risk of the genetically modified barley line PAPhy07 is the spread of pollen and seeds, which can cause unintended mixing with non-GM seed lots. We will apply the following risk measures to eliminate the spread of pollen to sexually compatible barley fields and seed dispersal to other fields.

The release site of the GM barley PAPhy07 will be situated 100 meters from the nearest barley field. A 2.5 m wide border of non-GM barley will surround the release site to minimize pollen dispersal. This border will be hand-harvested and all plant material will be destroyed at harvest. The sowing machine used for sowing the GM barley will be thoroughly cleaned and inspected for seeds. The GM barley will be hand-harvested to reduce seed loss. All plant material except the GM-seeds will be collected in plastic bags and burned. The spikes with the genetically modified seeds will also be collected in bags and transported to an indoor threshing machine. This machine will after threshing be thoroughly cleaned and inspected for seeds and waste material will be burned. After harvest, the release site will treated with Roundup to kill emerging seedlings. The next year, the release site will lie as black fallow and will periodically be harrowed and sprayed with Roundup. The subsequent year, cereals will not be grown at the release site.

Moreover, the release site will be checked regularly during the period of release. This will be done by visual inspection of the status of the PAPhy07 barley line. In the unexpected case of any adverse environmental effects observed this would be acted upon and reported immediately to the competent authority.

A report will be made available by the notifier at the end of the field-testing period. If any unexpected adverse environmental effects are observed, these and the further actions elicited, as an effect of these findings will be detailed in the report.

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Not applicable.
However, any unanticipated adverse effects on human health or the environment would be acted upon and reported immediately to the competent authority.

Final report
-

European Commission administrative information

Consent given by the Member State Competent Authority:
Yes
24/05/2012 00:00:00
Remarks:

Notification report