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General information Notification Number B/ES/18/03 Member State to which the notification was sent Spain Date of acknowledgement from the Member State Competent Authority 29/01/2018 Title of the Project Biomass production assay in tobacco genetically modified with the adrenomedullin gene Proposed period of release: 01/05/2018 to 31/10/2018 Name of the Institute(s) or Company(ies) Biomass Booster, SL, Muro de la Mata 5, 3º, 26001 Logroño, Spain.
Is the same GMPt been notified elsewhere by the same notifier? 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
tobacco solanaceae nicotiana nicotiana tabacum tabacum
Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications: A single gene has been introduced, namely mammalian adrenomedullin, under the regulation of the 35S promoter, present in commercial plasmid pBI-121 Genetic modification 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: A 570 bp DNA fragment containing the adrenomedullin gene was obtained by PCR from a human (Homo sapiens) cell line. This fragment was inserted in commercial plasmid pBI-121 substituting the GUS gene to form plasmid pBI-AM.
The intended function is to increase the plant´s biomass
Brief description of the method used for the genetic modification: The pBI-AM plasmid was inseted into Agrobacterium tumefaciens bacteria by electroporation. Small fragments of tobacco leaves were infected with the bacteria and then selected by antibiotic resistance. 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 Purpose of the release: To test in a field experiment whether the transgene increases plant´s biomass. Geographical location of the site: Experimental Station Aula Dei, in Zaragoza (Spain) Size of the site (m2): 3.250 m2 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: Not applicable Environmental Impact and Risk Management Summary of the potential environmental impact from the release of the GMPts: Environmental impact is not expected since no compatible species are found in the surrounding area and flowers will be eliminated to avoid pollen dissemination. Brief description of any measures taken for the management of risks: Weekly, the EEAD-CSIC technicians will check the healthy growth of the plantlets. Any time they detect an anomaly they will notify the principal investigator.
Once the harvest has been accomplished, the EEAD-CSIC technicians will check weekly for the apparition of new tobacco plants, which will be destroyed when found. Nevertheless, since flowers will be eliminated during the growth cycle to avoid seed formation and the plant remains will be triturated, buried, and watered to favor potential germination, and the field will be treated with herbicides, no problem is anticipated. In any case, any anomaly will be notified to the principal investigator.
Every plant and flower eliminated during the test will be triturated and buried, following the plan for remain destruction after liberation as explained in the previous paragraph.
Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release: Basically, this is a randomized complete block design with 4 replicated per genotype (control vs transgenic).
Each experimental unit is made up by a row containing 12 plants.
Distance between rows will be a minimum of 122 cm and distance between plants in the row is 56 cm.
From the laboratory, plants will be sent in small pots, ready for transfer to the field.
Dates for carrying out the experiment: between May and October. From transfer to harvest we expect a total of 7-8 weeks.
Although tobacco plants tend to auto-fecundate, flowers will be eliminated before they reach maturity to avoid potential polinations.
To perform the analysis, plants in the outside will be eliminated and only the 10 innermost plants will be analyzed.
At the end of the cycle (when all the plants had produced flowers), the following parameters will be measured:
1)Total plant height
2)Number of leaves per plant.
3)Length of the largest leaf of each plant.

Then the whole plant will be uprooted and the following parameters will be measured:
4)Fresh weight of the root, stem, and leaves of each plant. One leaf per plant will be frozen in liquid nitrogen for biochemical analysis.
5)All plant parts will be dried and weighed again.
Final report - European Commission administrative Information Consent given by the Member State Competent Authority: Not known
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