Several of the Arbequina explants are responding to the 427 protocol, however, the response has been taken considerably longer than the Tosca varietal. This explant was placed in new clear media today, and I can just make out adventitious roots developing on the stem.
Also today I started two additional Arbequina explants in an experiment to see if I can speed up the response. One explant is in the 427 protocol, while the other is in a protocol I will call 623. The 623 protocol contains more nutrients and an additional rooting hormone. This should be an interesting experiment indeed.
Two of the Tosca explants have been moved to stage 4 acclimatization. The plants may not have been fully developed and the move may have been premature, as some of the tiny leaves appear to be dying, however the stems are a healthy green and it may resolve itself.
One of the Gardenia vetchii explants had a nicely developed root system, so it moved it to stage 4, acclimation.
Using a sterilized mixture of peat and perlite, the plantlet was potted and placed into a domed container, to gradually get used to living in a normal environment.
Also today, explants were taken from Taggiasca and Koroneiki olive trees to be micro-propagated. So far, all of the olive explants in culture are responding to various degrees, but responding none the less.
A small section of branch was removed from the Picholine olive for cloning. After it was cleaned and rinsed the section was cut into three single node pieces that were then placed into culture media.
Rather than placing each explant in an individual jar, I have elected to use a single jar to save space and media. When, and if, the plants begin to respond they will be placed in individual jars for rooting.
The Tirilye seedling is almost entirely acclimated to a normal environment, at this point it is spending about 75% of its time in the greenhouse.
Today was transfer day for all of the olive cuttings taken on 4/27/14. A new batch of my #427 protocol was prepared, without any coloring this time. I have learned that coloring works for visually identifying the protocol, however, it makes it difficult to see what is going on beneath the surface. If coloring is added, it should be very light, just enough to distinguish the color.
Above is photo of a Tosca olive cutting that has now been in vitro for about seven weeks. At this point I regret that I did learn micro-propagation techniques much sooner and did not waste time, or resources, on traditional cloning. As is clear in the photo, the explant is developing a very nice root system.
Two ounces of protocol were added to each jar to give the plantlets plenty of room to develop a root system during this phase of the process.
The next phase of the process will be to transfer the plantlets to a sterile mixture of coir and perlite and begin the acclimatization process.
As of today I have several varieties responding to tissue culture, some better than others, however, I am certain at this point that all will be successful.
Since I began collecting olive trees I have been trying to locate a Picholine olive and I have finally found one. For whatever reason this variety is hard to find in this country.
Picholine Olive Trees are native
to France, they have large, flat, light green leaves, and medium sized
fruit, which can weigh about 3-5 grams. The olives are harvested green,
for eating, and black, for oil. They ripen in late November to
December. Picholine Olives are self-fertile and are known to be
resistant to both drought and cold, and can adapt to a variety of
temperatures and soils.
The tree I received had two trunks, one of which was removed, as I do not want any twin trunked trees.
The trunk that was removed was dipped in Vita-Grow rooting hormone and planted in Ava's garden with her corn. I thought I would give it a chance to root, and I did not want to root it using conventional cloning methods. My thinking is that the Roman army planted millions of trees, and they must have pretty much just stuck them in the ground as I have.
My previous attempts at cloning olives using traditional cloning have met with dismal results, which is another reason I just stuck the branch in the ground. Using traditional methods requires misting several times a day, without misting equipment the success rate is pretty near nil. I much prefer my present method of cloning using tissue culture.
The Gemlik seed was growing so quickly I had to make a decision concerning whether to move it to a larger vessel, or try potting it and acclimating it to life in a normal environment.
Obviously I chose the latter, as seen in the photo. Going forward I will use a larger vessel and more media when germinating olive seeds, as I did not realize that olives have enormous tap roots. The tap root had reached to bottom of the jar and began to push the entire plant well out of the media, so action was required.
A media of half perlite and half coir was prepared and a small amount of nutrient solution was added to the media. After placing the media into the pot, I sterilized both the media and pot at 121 degrees C and 15 psi.
The seedling was gently removed from its vessel and run under cool water for several minutes to remove all of the culture media. When the seedling was potted, the pot was placed in a plastic zip bag and the inside of the bag was sprayed with sterile water.
In the high humidity growing in vitro, the stoma on the leaves did not have to open to control humidity and turgidity, so it is necessary to slowly acclimate the plant to a normal environment. To do this, I will open the plastic bag for a short amount of time each day, and slowly increase the amount of time the bag is open for a period of about a month. At that point, the plant should be fully acclimated. During the acclimatization period I will light the plant with a 12 watt LED grow light using a photoperiod of 16 hours.
