Each year Oslo gives London the Trafalgar Square Christmas tree 'as a token of gratitude' for its support during the Second World War. Every year, Christmas trees are culled and thrown out after the festive celebrations. This Christmas, C-LAB attempts to create a tree rather than killing it.
Our first step in the operation was to get hold of a cutting of Trafalgar Square Christmas tree. With a help of a lab colleague, and a member of the security team (on Trafalgar Square) we obtained a small cutting.
Back in the lab, I was preparing and locating a media to tissue culture the cuttings. 200ml Murashige and Skoog media was prepared with 1.5% agar. The media contained supplements of IAA, Kinetine and Sucrose. IAA being an auxin hormone that, in theory, should induce root formation. The media was then autoclaved.
Arriving back with two types of cuttings (the Christmas tree and another plant we couldn't remember the name of) we were all set. But how do you clone it?
We had little experience in going about this - so we consolidated a plant expert: "Well, if you just want to clone it - it is a fairly simple process and more a case of gardening then anything else", he explained while scurtinising the cuts. "For tissue culturing, the problem is always to kill the microorganisms without killing the plant...but what you will need is a root inducing substance". We explained about the IAA, but he suggested adding an artificial version as autoclaving may render it useless. He took us to his lab and lent us a bottle with a pink powder - Indole-3-Acetic Acid or HetroAuxin or IAA (a child of many names).
Plates and tubes were poured with our plant agar. Then, we moved on to preparing the hormone stock. However, this proved tricky as the powder had a strange reaction with water and alcohol. In ethanol it quickly dissolved but as soon as water was added, it began to precipitate. In the fridge it formed a sort of jelly - all very strange behaviour for something soluble in water....
Dilutions of IAA was prepared. And we moved all material to a laminar hood that was cleaned with ethanol.
We cut the needles of the Christmas tree and tried to dissect them in the middle - this proved tricky as they are very tough. We used bleach to sterilise all the plant material and washed them for 10 minutes. The Christmas tree strangely bubbled - perhaps oxygen from the wood?
Cuts of the Christmas tree, its needles and the unknown plant were then placed in eppendorf tubes of diluted auxin or IAA and we put some in very concentrated auxin before placing them on the agar.
We debated whether they should be stuck into the agar or lie flat, and decided to try all possibilities.
A nice end cutting of the Christmas tree was place at the centre of one petri dish - and will hopefully pride it with roots.
Contamination, life or death - the coming days will tell.
