Thursday, January 16, 2014

Journal January 16, 2014 - Adding more samples to tissue culture trials

The initial olive cuttings have been discarded, as I did not believe that the culture contained the hormones required for olives.  Two different culture media, containing slightly different combinations of hormones, were prepared with food color added to differentiate between the formulas.   They are basically identical, except the blue culture contains IBA and Super Thrive.  

During the last two days new cuttings were taken and placed into culture.  The varieties include: Arbequina, Pendolino, Amfissa, Cailletier, Koroneiki, Taiggiaca and an unknown cultivar. 

As a further trial, cuttings from Jasmine and Gardenia were taken and placed into a basic establishment culture.  In only two days I have noticed that the Jasmine cutting is beginning to grow.

The vial forth from the left contains a section of California Redwood that is growing so quickly the growth is noticeable on a daily basis.  That said, I have no idea of what I am going to do with a Redwood in New York, other than attempt to bonsai it.

While viewing a video on tissue culture the presenter mentioned using tissue culture techniques to rejuvenate and germinate old seeds.  That prompted me to try the technique on olive seeds, though it should work on any seeds.  The presenter mentioned that the technique was used successfully on 20,000 year old seeds, so why not give it a shot?

Using a Dremel with a cutting disc I cut around the perimeter of the drupes and forced them open to release the embryos.

The embryos were soaked for one hour in 2 ml hydrogen peroxide to 40 ml water, with a single drop of detergent added to the water to break surface tension.  During this period they were agitated frequently.

Following the soak, they were rinsed in sterile distilled  water,  placed in a solution of 4 ml hydrogen peroxide, 40 ml sterilized distilled water, and 1 teaspoon of table sugar, where they remained overnight, or until the seeds had sunk to the bottom of the jar.

A 50/50 mixture coir/perlite was placed in a baby food jar and moistened with some of the sugar water mixture and a small amount of nutrients and 2 drops of super thrive. The medium was then sterilize in a pressure cooker for 20 minutes at 15 psi.

The following morning the seed was placed in the medium under sterile conditions. An aluminum foil cap was used to cover the hole in the jar lid and the container was placed with the other culture trials.

Also today, I noticed that one of the Koroneiki olive trees is beginning to flower.  The tree was removed from the winter chill area an placed into the grow tent under the 400 watt LED grow light.

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