This infographic demonstrates the simplicity of the Tissueponics cycle from plant trim to in vitro culture to plug ready rooted clone.
Prepare kit nutrients in a kitchen work space.
Additional materials required:
1. Canner size Pressure Cooker or 8 quart InstaPot
2. pH meter (suggested)
Create a clean space for tissue cutting to then place in nutrients to root.
Additional materials required:
1. HEPA air filter
2. Large clear storage box in a shallow container (still air space)
3. Clean grow room with shelf or rack system under low wattage lights
In collaboration with Ed Rosenthal on the newly updated Cannabis Grower's Handbook, we're featuring the Tissue Culture chapter written by Bill Graham.
The link is an excerpt from the fantastic book!
A: We started using tissue culture tricks for exactly those reasons back in 2008 and put all of the pieces together into the Seed kits. DNA and enzymes break down over time and the risk comes from aggressive fungus attacking the energy stores before the seeds can utilize them. We use TC preparation techniques to 1) sterilize the surface of the seeds, 2) hydrate them with a sterile germination solution with essential elements and vitamins, and 3) germinating in a sterile tube of soft coco medium.
Our standard kit contains ten tubes plus two tissue culture agar tubes to rescue seeds that pause in germination, plus tools and instructions. And there are refill kits to reuse the tubes. We can also make larger custom kits and provide materials a la carte for whatever you need. As with all new skills, you should learn by using on some expendable seeds and getting the techniques and timing down.
Check them out on microclone.com
This is the teaching Starter kit that is often purchased before the Complete Microclone Kit. Plant tissue culture is growth in a nutritional sugar media, the contents of the liquid media. You are likely familiar with agar-hardened tissue culture media which does not store and ship well and is a grower-mixed component of the Microclone kit.
The Starter kit allows you to perform your first plant sanitizing and media derived growth in culture and move into root plugs in only three weeks, two of three important steps to home tissue culture. The third is preparing media using a pressure cooker from the concentrates in the Microclone kit.
We make the complete kit for growers who want to learn and practice tissue culture on any scale and with nearly any plant. The Starter kit is made for growers who want to try before spending $230.
The tools and tubes are all used over and over with either replacement liquid media or Multkit concentrates.
The direct to rooting technique, also part of the Starter kit, is the foundation of the Tissueponics shortcut which is the most popular TC propagation method of cannabis and other growers with a regular supply of vegetative plants that are pruned.
A: Aquatics are most often raised on multiplication media containing branching hormones, When transplanted onto Root media like ours in larger containers to develop the roots they need to anchor. We have several users who have bought aquatic plants online already in sterile culture and have split them into hundreds and even thousands of new little plants they then move onto rooting media with agar, the components in the Rootkit.
A: Yes, you would need agar to solidify. We can include the agar at no extra charge and the media would have to be resterilized in a microwave or pressure cooker if you have one. The Tissueponics media is shipped as a liquid because it is it sterile, easy to handle and pour, and does not spill or break like the agar media we used to ship in tubes.
To sterilize in a microwave, fill a one liter measuring cup with 250 ml of water and heat to boiling three times, allowing to boil about 20 seconds and noting the time. Empty the water and do the same with the Tissueponics media with agar added and dispense into tubes. Do you already have the tubes from the Tissueponics kit?
A: It will outrun HPLVD in every case so far. Viruses are always chasing the tips and the small tips taken for culture outgrow them. You are cleaning a system, not necessarily individual plants. the strategy is to top the best plants in the veg tables and put those small pieces directly onto rooting media in shortcut tissue culture. After a few cycles, old mom plants are no longer needed and all plants have been through disease identifying tissue culture. If you see disease, dump it and replant it. There are plenty of clean plants to make each new cycle.
Selecting frequently for vigor and other desired characteristics usually does the trick, along with moving veg plants into flowering no later than six weeks so all plants are replaced with fresh new ones, like your skin. Wait about two months to do any testing and even then, it is to see if ANY virus exists in the population.
Our testing method is to make short clones from the best plants and to send in 3/8" cut from the bottom of the stem when making the 45 degree cut. Viruses cannot grow past stem tissue so negative tests mean there is zero virus in the rest of the cutting. The short cutting, or tissue culture cuttings, mean the stem sample is soft enough for digesting and buffering. Every one of our growers who have sent plants to test using our method have come back 100% virus free.
Viruses are another reason to not keep mother plants that get over a few months old and are the exclusive source of clones. Turnover, regular clean evaluation, and frequent selection keep the system clean and are part of routine Tissueponics.
Also once the media jars (baby food) are sterilized and filled is there a reasonable shelf life?
A: Hello, and thank you for asking. Most houseplants are tropical and prefer the yellow BA Multkit, also listed on eBay. Larger philodendrons, particularly Monstera, clone well in culture, but cutting up donor plants and getting then cleanly into culture is the challenge. You will need to carve out the growing points behind the leaves and clean them completely with a soft brush and bleach. The media jars are good for a while if stored carefully in the same clean area the plants will be grown in. We have used media that is 6 weeks old with only a few spoiled containers from storage. If you know you are not going to use the media for a while, store the unprepared media in the fridge or freeze if sugar has already been added, until closer to the time you will need it.Look up the cutting procedures published online and let us know when you will need media or a complete kit with tools and bottles.
A: Thank you for asking. TDZ is more often used for woody plants but is worth trying for the plant you have. We have three different Multiplication media to try-BA, TDZ, and mT. They will all work but one may work better for you and your plant.
We make a cannabis kit and added TDZ for users wishing to copy a popular published research paper.
You may also be interested in our exclusive Tissueponics kits which skip multiplication altogether for faster results and better success. It works by rooting clones in tissue culture from small tips and nodes pruned from veg plants. See the videos on Youtube.
Use cloning best practices with an extra emphasis on humidity and air circulation. We use tall domes with the vents open inside of a 75% humidity chamber. Burp domes a few times a day to encourage hardening and spray with 0.1% H2O2 to keep opportunistic fungi away.
A: Excising seeds in tissue culture. The excising kit is part of the Seed germinating kit, as a secondary "rescue" technique. Anthony can separate an embryo from its seed coat after thorough cleaning and hydrating with the kit's solutions to loosen the seed coat and minimize the damage to the embryo. The longer he can wait, the better, especially to wait until the seed has shed its coat naturally.
He may want to soak twelve seeds, plant ten in the sterile tubes to germinate normally and leave two in the hydrating vial to shed their coats. Tell him to drain the hydrating solution after removing and planting the ten tube seeds. The thin film of sterile water will be enough to allow seeds to push out their root radicle, swell and loosen their coat. Infact, the coat in the culture tube makes little difference except perhaps if it carried some small contamination.
The growth is going to come from the primary meristem as normal, and maybe from a cotyledon or part of the seed stem. Anthony will need to move up the the complete Microclone kit when the seed begins to grow in culture.
Check them out on microclone.com
Q continued: My goal with the kit is to clean up some old clones and attempt meristem tissue culture on them. I'm just not sure which solution I should use to start. In your videos you are using what looks like the yellow BAP kit solution. My kit only came with pink colored starter root kit solution. Is that all I need for now?
I also had a question about the chlorine cleaning solution. I want to make my own solution when I run out of tablets I just don't know the ratio being used in the solution. Right now I make my own "Clear Rez" solution for my cloner. I use pool shock which is 68% calcium hypochlorite. Is there a formula for the solution I can reference.
A: The best way to meristem plants using TC kits is to learn and set up your TC area based on either the 16 tube Starter Kit you are looking at with several follow-ups using replacement media listed on eBay, or as most users do to get the larger Complete Microclone kit for $219 and having the media and containers for several meristemming rounds from the get go.
The most successful strategy is to learn TC using the Tissueponics shortcut Starter kit, putting 2 inch mini cuttings directly onto Rooting media, moving out of culture into root cubes after three weeks. First, Tissueponics is surprisingly successful at eliminating suspected virus infection even with two inch cuttings. To pursue smaller meristem cuttings afterward, you will be glad to have proven grow areas and cuttings techniques. Progressively smaller tip cuttings are either taken ex vitro and chlorine sanitized to go into culture on Mult media, or taken from plants growing in Mult media culture already, usually both.
We strongly encourage both disease elimination and production growers practice the Tissueponics clean plant cycle and avoid keeping mother plants older than eight weeks. TC tubes, young TC-derived veg plants, and their incidental clones can all be kept effectively clean together in the same closed, HEPA, controlled-traffic environment.
We use one to two grams of Dichlor which makes a 500 to 1000ppm solution of free chlorine. Use the 99% sodium Dichlor powder to avoid the non-active ingredients.
How many plants do you produce, how often?
A: Start with 36 pieces max of each strain while learning and to get the roll. Fifty-plus minutes each wash. Cut and rinse a few batches ahead of time and stir in Dichlor as each previous batch is heading into tubes.
Easiest to use multiple wash containers lined up. Don't get ahead of yourself. I have seen many oversoaked plants later spoiled in tubes. Use a few kitchen timer from the Dollar Store and set one for each soak.
I use the same technique for large daily runs and the timers make a huge difference. You will soon speed up to two or three plants every other minute and will be able wash more plants during the next plants 50 minute cycle. I use fifty minutes on the timer because it is only a few minutes later that the first plants in that wash are going into tubes and most go into tubes closer to 60 minutes, minus or plus.
I label the first plants going into the rack, and the last ones, and can tell if there was any difference. Most often, they are fine and I will begin cutting minutes off the washes at later times.
You may also increase the strength of the Dichlor to two grams per liter to reduce wash times. The one gram in the kit is calculated to the 50 minutes newbies take to get plants into a 36-site tray. I wash and stick 36 Tissueponics cuttings in twenty minutes directly from a 2g/liter soak and put in a new batch when I grab the last one and use a single timer set to twenty minutes.