Tissue culture and cannabis?

GrowerGoneWild

Well-Known Member
You can always order the agar media and some come with anibiotics in the mix to prevent or prolong micobial growth while you administer the growth hormones along the way. As I'm sure you already know, there's multiple hormones needed for a TC, and each has a specific function. Grow roots, grow stems, and leaves. Orchid enthusiasts and forums alike have a good deal of info on TC and give reputable retailers for the equipment you'll need.
PPM (Plant Preservative Mix) is the biocide I will be using for the TC I dont know if it also has antifungal qualities. I'm going with a generic MS medium with vitamins.

The cytokinin's and auxins will be mixed into the agar.. and will be done in stages. Rooting agar will be a different formulation, and will be marked with a different color.
 

Keighan

Well-Known Member
You have a $200,000 lab complete with centrifuge, etc?
Just saying dont care the reprecussion from this you are a very intelligent grower and alot have learned alot from you but youre seriously one of the few sore thumbs in this stoner idiolistic society we like to think of it as going against the grain and being a conceited douche frankly, ive seen you do it on tons of threads to people who didnt deserve it and werent talking about or to you, cant everyone just be peaceful, informitive, and helpful?...really
 

Grandpa GreenJeans

Well-Known Member
Just saying dont care the reprecussion from this you are a very intelligent grower and alot have learned alot from you but youre seriously one of the few sore thumbs in this stoner idiolistic society we like to think of it as going against the grain and being a conceited douche frankly, ive seen you do it on tons of threads to people who didnt deserve it and werent talking about or to you, cant everyone just be peaceful, informitive, and helpful?...really
You forgot supportive. Lol just sayin.
 

Grandpa GreenJeans

Well-Known Member
My bad I was trying to explain my frustration without appearing as rude. Or atleaste as respectful as I feal I could be.
I wasn't put off by it. You make a very valid point, that were all here to share teach, help and support each other. Cannabis has a weird way of bringing people together and we all share the same passion for the plant. Sometimes it's good to remind people politely were all here for the same reason. Even though old timers (like myself included) have been known to have inflated egos, we're all the same.
 

Keighan

Well-Known Member
And the new generation like me is beyond grateful for all of your "old timers" knowledge, Im constantly throwing my pride in the trash and asking questions or asking the local guerilla growers. Its a beautiful experience..new age og.
 

Grandpa GreenJeans

Well-Known Member
I got lots of 1/2 pint jars, but I'd like to use Polypropylene containers (PP) cheap and plentiful, and they can be sterilized. and the top is transparent, I figgure I could get about 5 to 10 explants into the container.
Mushbox.com has the containers I believe. I've bought from him 5 years ago and still have the PP jars. If your into mycology and isolate your mycelium on petri, I have had good luck with his nutrient agar. Pre made and sterilized.
 

GrowerGoneWild

Well-Known Member
Just saying dont care the reprecussion from this you are a very intelligent grower and alot have learned alot from you but youre seriously one of the few sore thumbs in this stoner idiolistic society we like to think of it as going against the grain and being a conceited douche frankly, ive seen you do it on tons of threads to people who didnt deserve it and werent talking about or to you, cant everyone just be peaceful, informitive, and helpful?...really
UB's comment is so far removed from reality.. 200K to make cannabis tissue clones?. centrifuges?..

He does have some knowledge as a gardner, but when he says stuff like that it makes him sound like a keyboard warrior.

Anyways, we are all just gardeners, and farmers.. with different viewpoints, so no real reason for anybody to get a chip on their shoulder. But then again this is "troll it up".. More effort towards dissrespect than discussion.

Anways back to Tissue Culture :)
 

Grandpa GreenJeans

Well-Known Member
I'm going to start off with some fan leaves, cut that up into its natural sections, and use small shoots from the main stem that would be deemed too small for clones.
That sounds like a good start. You can virtually take the tissue from anywhere on the plant for a TC. I have seen alot of orchid cultivators take the inner most tissue. They target new growth, i guess its because it has the highest concentration of growth hormones.
 

coconutbeach

Well-Known Member
Mushbox.com has the containers I believe. I've bought from him 5 years ago and still have the PP jars. If your into mycology and isolate your mycelium on petri, I have had good luck with his nutrient agar. Pre made and sterilized.
Four and six ounce polypropylene and glass jars are best for gel culture. We are also using 23mm plastic tubes for introductions and starts. All are included in the Microclone tissue culture kits. planttc.com
That sounds like a good start. You can virtually take the tissue from anywhere on the plant for a TC. I have seen alot of orchid cultivators take the inner most tissue. They target new growth, i guess its because it has the highest concentration of growth hormones.
Make it easy and use 1-1/2" clean vegetative branch tips.
 

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coconutbeach

Well-Known Member
UB's comment is so far removed from reality.. 200K to make cannabis tissue clones?. centrifuges?..

He does have some knowledge as a gardner, but when he says stuff like that it makes him sound like a keyboard warrior.

Anyways, we are all just gardeners, and farmers.. with different viewpoints, so no real reason for anybody to get a chip on their shoulder. But then again this is "troll it up".. More effort towards dissrespect than discussion.

Anways back to Tissue Culture :)
Startup cost of Microclone tissue culture. Kit, pressure cooker, tub, rack lights. Less than $400 for three months growth and first 150 rooted clones.
 

coconutbeach

Well-Known Member
Meh, I've run sugarless mediums like in the case of psylosybin cultivation, you can have contamination take hold if the medium has sugar in it or not. It might decrease the likelyhood of infection, so point taken.

Actually the method is not like mushroom cultivation, the byproduct of fungi is CO2, and it does not take much airflow, in fact on the Brown Rice Flour (BRF) cakes I make I do not expose them to circulating air until a fruiting body shows.

So back to "photoautotropic propagation," the idea is that CO2, is the food versus Sucrose in the agar. If we look at Chieri Kubota's work she says that in vitro propagation is slow due to the limited ammount of co2 in vitro. And I somewhat agree after looking at the time it takes from Stage 1 to plant hardening in Wang's or Scroggins work on in vitro cultivation. Off the top of my head from explant to hardened plant takes somewhere around 6 weeks.

Well at the very least we could flood a laminar booth with a high level of co2 when inserting explants into the desired medium. In Kubota's work it looks like there is more mass vs regular explants. And needless to say co2 we know is a basic building block of mass. I'm now sure how I would implement this and mantain a sterile culture at the same time flowing co2 over explants in vitro, not to mention the cost of concentrated co2, as in a compressed cylinder might push me away from going this route.

The abstract you mention is wang's 2009 study on the micropropagation of hemp... Cannabis Sativa L. Same thing if you ask me as the typical cannabis hybrids. The success rate after Stage 3 to hardened plants is 99%.

Well I'm researching this now, I think I'm going to go commercial, and provide clones to commercial grow ops.. I'm looking at pushing clones out in numbers like 200-500 clones a month.

At the very least I've learned a bit, from Kubota's work, I could use the co2 to optimize growth in my explants. ;) I'll continue to at least look into the subject.

Like.. good stuff man...
Check out Microclone tissue culture kits at planttc.com or at Alaska Jacks in Palmer.
 

coconutbeach

Well-Known Member
Isn't diluted bleach economical? My understanding is that college labs teaching tc use flowers and use diluted bleach as well
A typical wash is 10% bleach for 15-20 minutes, however, we much prefer 500ppm of dichlor because the Cl evaporates and typically does need the additional sterile water soak. Dichlor is included in the Microclone TC kits.
 

coconutbeach

Well-Known Member
im gona try it at our new place im gona invest in all the steril equipment and just for shits and gigles im gona try to market them lolz who knows it might work as far as genetics go sounds like another project new place almost doneView attachment 3336537View attachment 3336538 View attachment 3336541
What a great space. I am in CO as much as I am at home these days. Your space can make all the clones you need and you will likely still have space left. You will need a filtered air area for growing and handling, kitchen for media prep and dishes, and rooting trays and racks. OUr biggest setups still started with a Microclone kit and scaled up from there. See how you can HEPA filter the ducted air at one end and curtain it off for TC racks.

These are the racks we made to fit the width of the four foot shelves. We call them Microclone Rack Trays.
 

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coconutbeach

Well-Known Member
it's basically for preserving genetics. If you want 1000 genetics in a small warehouse, tissue culture is where it's at. It works great. I haven't done this myself, but I know people who have. I wouldn't take a tissue culture for something you want to flower next month though, but it does have it's use.
Check out Microclone Cannabis Tissue Culture. Search Google.
 

coconutbeach

Well-Known Member
Ideally I want an incubator in addition to the same veg space I use now, so just eliminating a dedicated mother space for ~20 strains I'd wish to hold on to. Does that sound like a reasonable application here? Glove box, filter etc are within reach.
The first Microclone grow racks held multiple vessels of over 30 varieties on one 2x4 ft shelf under two 54-watt t5's.
Nice job thump! I look forward to more info.

I've seen attempts and journals at several forums and they typically end before it becomes interesting.

Some links I bookmarked, can't vouch for the quality, but about cannabis specifically:
http://www.lycaeum.org/mv/anagrams/cannabis_tissue_culture.html
http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf

That's the downside of tissue culture many don't realize. Regular cloning is much and much faster.
Regular clone ROOTING is much faster because of the resources put into creating the 6" + cuttings. We still always have some rooted clones to back up the cultures and prove the vigor. It is all of the growth, potential, and advantages that tissue culture presents that should be explored and utilized. Tissue culture is basically growing plants with sugar. The rest of the items we love are there too like minerals and vitamins. Using sugar means we don't need roots to grow plants and create new cuttings but that we will also need to prepare clean media, clean plants before introducing into culture and handle with clean tools in a clean area.

Before I scare anyone by using clean so much, I regularly do tissue culture on the road in hotels, countertops, and on folding table demonstrations at hydro and cannabis trades shows. Tissue culture is how the commercial horticulture industry clones nearly all of their plants and the advantages are ideally fitted to cannabis. Hundreds of clones are produced from only 8 sf under 100 watts of light every month. And we typically stack those racks five or six high for the big hitters. One outfit has even added another three feet to their wire baker's racks to go to nine feet and eight shelves per rack.
 

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