Tuesday, February 25, 2014

Journal February 25, 2014 - Kalamata seed in vitro

The photo above shows the Kalamata olive seed I placed into culture on January 26, 2014.  In less than a month it has begun to grow, however, it looks nothing like any seedling I have ever seen.

It looks more like an ugly green slug with yellow eyes and green eyeballs than it does like an olive seedling.  Perhaps I got carried away with the hormones.  

Well, in any event, it is growing, and that is fine with me.  Four weeks is much better than waiting for a year and nothing happening.

The photo was sent to the Tissue Culture expert who has been giving me advice and she replied that it appears callus is forming on the tissue.

What is callus?  According to my book: "Callus is a mass of undifferentiated cells.  The callus mass can contain embryoids capable of developing into whole plants or it can contain shoot or root primordia. (the earliest developmental stage of an organ or cell)"

Who knows?  I may get more than one plant from this seed.  Only time will tell.....

Saturday, February 22, 2014

Journal February 22, 2014 Manzanillo in vitro

The GA7 vessels I purchased from Kitchen Culture Kits are really nice to work with.  Being larger vessels they allow you to work with larger explants.  Positioning the explant in these larger vessels is a breeze. 

The above photo is my first real attempt to use a woody cutting for an explant.  The cutting was taken from my stubborn Manzanilla olive.  The plant is finally beginning to put out some new growth, so I removed a growing section from one of the top branches for this trial.

After cleaning and sterilizing the cutting I scoured the base of the stem before placing the explant in the protocol.  The protocol contains an abundance of a rooting hormone, IBA 

As misting traditional cuttings to maintain humidity is a real problem without automatic misting equipment, my thinking is that the GA7 vessel may solve the humidity problem.  

If you look at the cap on the vessel you can see small ribs that allow air exchange, and, they do a great job of controlling the humidity also.

Thursday, February 6, 2014

Journal February 6, 2014 - Arbosana culture test

The Arbosana olive tree was too tall and needed to be cut back, so I decided to remove about six inches from the top of the tree to be used for a micro-propagation test.  This would be the first time I have used woody material rather than soft green growing shoots.

The section removed was surface sterilized in H2O2, cut into several single node explants which were placed into a variety of protocols.

There are things going on with the current tests, however, at this point, I do not have enough experience with the process to interpret what is going on.

For instance, five Kalamata olive embryos were placed into culture on January 6, 2014.  On the morning of February 2, 2014 I looked at the embryos and could see swelling.  That evening one of the embryos had burst and white tissue was protruding from the blunt end.  As of today the tissue is growing in a downward direction and growing quickly.  Normally, you would expect a single radicle, pointed like a finger, but this mass of tissue looks nothing like a radicle.  I have no idea of what is going on at this point.  There are four other Kalamata seeds in the same vessel which are swelling as well.  

Previously, I have tried to propagate olives by germinating seeds, but even after trying for a year had no luck.  To actually see something happening in eight or nine days is exciting, even if I don't know what is happening.

Time will tell.....

Monday, February 3, 2014

Journal February 3, 2013 - Gardenia Veitchii in vitro

A small single node section of Gardenia Veitchii was placed into culture on January 9, 2014.  Today, the section was transferred from the unicorn vessel to a glass baby food jar with a polycarbonate cap.  

The glass vessel contains a standard MS medium with BAP.

After only three weeks in vitro,  four new plantlets are developing on the explant.  To say the least, micro-propagation is a fascinating process.