Because of the long generation time and obligatory out- crossing of most trees, reverse genetic approaches are the only realistic alternative for the analysis of gene functions in trees. UPSC scientists were among the first in the world to be able to create transgenic trees. All initial work was done with a fast-growing and relatively easily transformable hybrid aspen (Populus tremula x tremuloides) clone originating from the Czech Republic called “T89”. This is still the main tree species used in the transformation facility, but the numbers of different tree species that are routinely transformed have now grown to include elite varieties of hybrid aspen, Populus trichocarpa, Populus alba, Populus tremula, Betula pendula (silver birch) and Eucalyptus species.
The method used is Agrobacterium-mediated plant transformation of stem segments, petioles or leaf explants and the facility is today growing more than 15 000 transgenic trees, representing more than 3000 different lines, probably making this the largest tree transformation facility in the world. Through an agreement with the forest biotechnology company SweTree Technologies, the facility is gradually building up a biobank that will eventually contain transgenic trees representing not only the above mentioned trees, but also trees with down-regulation or overexpression of 2000 different genes potentially involved in the regulation of wood formation, providing a globally unique resource in the world for future research.
The transformation facility currently employs six persons and is servicing all of the research groups at UPSC and collaborators at both Umeå University and the Swedish University of Agricultural Sciences. Any group requiring transformed trees simply needs to provide the Agrobacterium strain carrying the genetic construct they want to analyze. After 8-10 months the trees are ready to be transferred to the greenhouse for analysis.
The persons working at the facility are:
Detailed Procedure information
To request a transformation you first need to fill in the form found under request transformation which you find here to the right. Please follow the guidelines stated there.
You then provide the Agrobacterium containing the construct of interest as a pellet from a 50 ml liquid growing culture with an OD between 0.3-1.0 (max 1.5) in the morning of the given transformation day (preferably before 9.30 am).
The facility will then do the transformation (3 days),
selection on plates (2-3month),
growing the callus (1-1,5month),
growing the shoots (1.5-2month)
selection of the shoots to get the individual lines (1 month).
If you forget your transformation appointment we will charge for 1 hr work (400 SEK) (this is the time for us to prepare for the transformation in vain and putting everything back)
Selections available Kanamycin 80 µg/ml
We can do transformations all year long but during the summer (midsummer to mid august) we do not redo the transformations for free if they get infected.
Download the pdf here to the right and fill in the request of amplification, print it out and put it in the basket on the 4th floor. State both how many plants you want and also how many you want in each jar, the default is 3 plants per jar. Also remember to fill in if you would like wild type plants, and in that case, how many. Usually when starting an amplification we only have 3 shoots to start with, so depending on how many replicates you want it might need several rounds of propagation. One propagation round takes normally between 4-5 weeks to grow. So please plan ahead. We will try to inform you when we start the amplification and when we expect it to be ready.
We will inform you when your transgenic lines are ready to be tested. There are two possibilities:
We can take the samples (stem or leave) for you as soon as the first individual lines are ready and freeze the samples in liquid nitrogen.
The price for that is 35 SEK per sample
The other possibility is to wait a few months more until the next routine cut and then you can get the old jars.
The disadvantage of that is that we have less material to start with if you order an amplification of your selected lines.
Price list (PDF format)
Online request form