Tuesday, January 28, 2014

Journal January 28, 2014 - Seeing some progress

Yesterday I separated the  Redwood explant that I received from California.  The single plant has turned into four plants in only a month. The separated new plants have been placed into culture to make even more plants.  The Redwoods are in the small vials on the right in the clear protocol.

Doing the math, at this rate, the process would produce three or four thousand trees in about a year. 

Today I took a branch tip from the Pendolino olive tree, sterilized it and cut it into three sections of equal lengths, each with two nodes.  The sections have been placed into three different protocols each containing different constituents.  As all three vessels contain sections of the same branch this should be a very interesting experiment.

The more I work with this process, the more I find I like and enjoy it.  It really takes up very little space, needs very little light, and, needs practically no maintenance.  Each plant is in its own little self contained environment; needing only to be moved to a new vessel every four to six weeks.

Sunday, January 26, 2014

Journal January 26, 2014 - More tests

A day or so ago I read an abstract on micro-propagating olives that stated they rooted easily in a protocol containing one half MS with vitamins and a hormone, NAA.  That got me really curious, as after all I an not so much multiplying them by the hundreds, but rooting them reliably.

On 1/24/14 I prepared a protocol with 16 ounces of distilled water, half strength MS, .5 ml NAA, .5ml PPM, 1.5 teaspoons sugar and 1.5 teaspoons agar, coloring it yellow. A Manzanillo cutting with two nodes was placed into that protocol as a test.

On 1/25/14, after reading a recipe on homemade protocols, I decided to go for broke and brew up a small batch of my own to test. To that end I used 1/2 cup distilled water, 1/2 cup coconut water, 1/2 cup of a stock solution of Expert Gardener plant food, with added Super Thrive and .5ml of Vita Grow, plus the following vitamins straight from the pharmacy: 1/4 tablet each of: Inositol, B-Complex and multi-vitamin.  That protocol was colored green and a similar Manzanillo cutting was placed in that protocol as a companion test.

It should be interesting to watch, as my homemade protocol has enough vitamins and nutrients to grow hair on a brass doorknob.

Monday, January 20, 2014

Journal January 20, 2014 - Cloning Arbequina

Of all of my olive trees, the Arbequina olive, shown above, deserves to be genetically duplicated.  This tree, although less than a foot tall, has flowers developing nearly every branch.  I doubt if every flower will set an olive, but even if a quarter of the flowers set, I fail to see how such a small tree will be able to support them.

The buds are starting to open and I am finding small patches of pollen on the leaves below the flowers.  When the flowers first open the stamens are closed, but open a short time later and apparently spew pollen out.  Using an artist's brush I am daubing the tip in the pollen clumps and transferring pollen from flower to flower like a bee.  That is probably not necessary, but I really want to see how productive this little tree can be, so I am taking no chances.

Today I removed two sections of growing tip, each about 2 inches long, surface sterilized them and placed them into culture.  As I wrote above, this little tree deserves having its superior genes replicated as many times as possible.

Sunday, January 19, 2014

Journal January 19, 2014 - Plants From Test Tubes

Today I found what I think is the perfect photo to decorate the fronts of my tissue culture light boxes.  That is boxes plural, as I am finding that one small box has no where near the capacity that I am going to require.

The box shown in the photo above will be illuminated by only a 125 watt equivalent 5500K CFL, as really not much light is required.  This box will also serve as my "still air" container when preparing sections for culture. The second box, containing the current batch of cultures, is now under the 150 watt Reflector LED grow light using only the vegetative setting.

My copy of Plants From Test Tubes, by Kyte, Kleyn, Scooggins and Bridgen, finally arrived, and it is an excellent book for beginners like myself.  The book has changed my perspective on the process somewhat, and this could become a hobby to equal hydroponic gardening.  It is a fascinating pastime, and, as the plants are sealed in their own environment; they require very little maintenance, taking up not very much space.  

So far, two olive varieties are showing signs of growth, and my initial trial cuttings are doing really well.  The Jasmine sambac has at least five plantlets beginning to grow from the single node placed into culture.  In 4 to 6 weeks the plantlets can be excised and placed into culture to form new plants.  When large enough, the plantlets are placed into a medium with rooting hormones and allowed to develop roots.  Following that stage they can be removed from culture, washed, placed into sterile growing medium and acclimatized in a protective environment.  According to the calculations in the book, if each test tube only produced a single cutting, in only 11 months you would have 2084 new plants.  Considering that my single node has already showing five plantlets, the total possible production is astounding to contemplate.  As there are three stages to this process that will require growing in culture in containers, I quickly determined that I needed more room to maintain the cultures.



Friday, January 17, 2014

Journal January 17, 2014 Cloned jasmin sambac

The way I got into tissue culture was by bidding on a kit on eBay.  As usual, I bid low and did not win, however, the person who did win backed out apparently thinking: this is not for me.  The seller wrote and offered me the kit for what I had bid, so I snapped it  up.  After all, it was going to be a long cold winter and I needed some distraction.  And, as usual, I was skeptical about the claims made regarding tissue culture, but I am now a believer.

The photo shows a single node of Jasmine sambac, Maid of Orleans, that I place in culture eight days ago.  From the single node there are now several new plants beginning to grow.

Further, I noticed this morning that a single node section of Taggiasca olive placed in culture three days ago is showing signs of growth.  Consider that I have been trying conventional cloning methods for almost a year with little success.

The Taggiasca section is in one of the blue vials shown in yesterday's post.  The red culture contains a combination of hormones I found in an abstract from Portugal.  To that combination I separated 480 ml and added .5 ml IBA, and three drops of Super Thrive.  Like, who knows what they put in Super Thrive?

So far, none of the other olive varieties have started to grow, but the Taggiasca may simply react differently.  To say I am delighted would be putting it mildly.

There are two people with sites that I found to be extremely knowledgeable and helpful, so if you have an interest you might like to visit them:

Plant TC  and Kitchen Cultire Kits

Additionally, there are several videos on YouTube that I found useful.

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.

Thursday, January 9, 2014

Journal January 9, 2014 More micro propagation trials

Having two vials of gel left from the initial olive trials I decided to try more common species. I am hoping that they may prove less difficult to propagate than olives so I can get a better idea of how this process works.

Single node cuttings from  Jasmine and  Gardenia were taken and placed in vials under sterile conditions.  The cuttings have been placed under the LED grow light along with the olives, which by the way still show no signs of growth.

Florist's tape was wrapped around the tops of the vials to keep out contaminates; I find it much less expensive and easier to work with than the tape supplied with some kits.

Tuesday, January 7, 2014

Journal January 7, 2014 Vegetative stage lighting

The Manzanillo olive tree has also been moved to a warmer environment with feeding resumed.  This tree was received in August, 2014 and has been in shock since then.  

It was literally plucked from a grove in California, had the top and branches chopped off, then spent a week in a box being shipped across the entire country.  No wonder it went into shock.

Finally, I noticed the tree beginning to put out new growth while under winter chill conditions.  It is my intention to get it growing again quickly, so it can perhaps make up for the several months of growth it lost.

The light being used is a 150 watt reflector LED grow light.  For now I am only using the vegetative stage, however, the tree is still receiving 3,000 foot candles with the light a foot above the canopy.  The photoperiod will be sixteen hours and the temperature will average about 74 degrees.  

These trees fruit early,  bear clusters of large olives, and I suspect that someday this tree will be one of my best.

Monday, January 6, 2014

Journal January 6, 2014 New 300 watt LED grow light

Surfing eBay two weeks ago I came across a new 300 LED grow light that was almost half the cost of the light I purchased a few years ago direct from the manufacturer in China.  And, the light was located in the United States, with free shipping.  Shipping from China was always an issue, with an average cost of sixty dollars on top of the cost of the light.

Although I did not really need another light, the price was attractive, and I could always use a spare, so I made an offer.  The new light arrived today and I placed it in one of the tents to try it.

At the height shown in the photo I measured slightly more than 3,000 foot candles at the top of the canopy.  Measuring the level outside the perimeter of the light case, I found that it has a very nice dispersion all around the case.

When dealing with this type of grow light, foot candles really only serve as a measure of intensity, not effectiveness.  The spectrum is equally, or more, important than intensity.

Description :
  • 9 Bands for Indoor Plants Growth & Flower
  • Spectrum of Light: 430~440nm, 450~475nm 620~630nm, 650~660nm, and white
  • Real IR spectrum: 730nm
  • LED Output Power: 100pcs*3watt
  • The range of power consumption under 110V: 182W-196W; 220V: 175W-184W
  • Dimension: 400mm*212mm*60mm
  • White painted casing.
  • Lifespan: 50000-100000 hours
  • View Angle of leds: 90/120°Mixed

Sunday, January 5, 2014

Journal January 5, 2014 Simulating spring conditions

On the advice of an olive grower I moved the Arbequina plant that is flowering into one of the tents today.  It appears that the tree has set all of the buds that it is going to set.  A warm environment, increased light intensity and duration, and resumption of regular feeding should hasten the development of olives.

For several weeks now I have been harvesting the outer leaves of the Garnet Rose romaine lettuce plants.  After each harvest the plants come back again and again and again.  Amazing.

Another of my current lettuce crops is red salad bowl.  I planted the salad bowl as it is not normally available in the winter, so it is a reminder of spring and summer.

Last, but certainly not least, I have a crop of Dutch Winter Brown lettuce growing.  This is a heirloom variety that was grown in colonial times by Thomas Jefferson in his gardens.  I believe the seeds cost less than three dollars; considering I can grow a few hundred plants that is certainly a bargain.

Wednesday, January 1, 2014

Journal January 1, 2014 Tissue Cultue Trials

Leccino and Cailletier cuttings were started today which consist of a 1/2" section of growing stem  with a single node.  The instructions I have been following said to remove all plant material, so that is what I have been doing, however, after reading a research paper on  mircropropagation I found that in the case of olives it is best to leave a small section of leaf.  I decided to leave just the small bud found on almost nodes.

Also today I removed the cuttings that had leaves, reduced them to 1/2" sections with a single a node and replaced them in vials.  I will be amazed if they actually take.

Prior to having read the research paper I thought that this was going to be a short process, but the paper said that their trial took 99 days for sprouting to take place. Following spouting the plantlets were moved to a generic media, where they continued to be grown until large enough to be transplanted to their final media.  Bummer...

They also used some growth hormones, which I am not sure if my culture media has.  Going forward I will mix my own media and add BAP, NNA, VitaGrow containing IBA and perhaps Super Thrive just to see what happens.

It will be equally amazing if any of this actually works, but it is a fascinating process, and that is what retirement is all about, keeping busy.