# Plant Tissue Propagation ?



## ozman (May 20, 2009)

Has anybody here tried tissue propagation and if so,level of difficulty,
yadda yadda .
But I have done some research on it looks like it would take time to get clones from but feasible I suppose.
Since I cant seem to clone anymore looking at options.


ostpicsworthless:

:watchplant::lama::bolt:


----------



## Sir_Tokie (May 20, 2009)

Why can't you clone? I believe cloning would be eaisier than tissue propagation. Look in the DIY section they have a DIY bubble cloner that is what I use with 100% success rate...take care..


----------



## ozman (May 20, 2009)

Been there done that,also built a aero cloner from the diy....
IM not sure what im doing wrong yet,
Ive been trying everything,from rockwool,and hydroton too bubble cloner,peat pellet pucks,I now have 2 jars of water sitting on the kitchen window sill with 10 cuttings in them.
Ive installed a reverse osmosis system 6 stage, didnt help,Im now buying water from a commercial water station RO water with a ph of 5.1 to 5.5
ppm is 11-18.
I keep my ph in the cloners,bubble or aero,at 5.5-5.8.
Ive tried scarification,splitting the stems, all the dips and gels and powders and liquids you could just about mention.
   My first try on the aero I used dip n grow when I cut my slips,they developed a gelatinous gel on the stems,hence the stem rotted,I have 1 out of 25 that has any resemblance of roots I placed it the jar on the kithcen window sill it looks ok for now.
I used to grow 20 years ago I had a 99% sucess rate, now in 8 months time ive gotten 11 to root and grow,I was using superthrive on those.

So i dont know it is dishartening.But I will go on and continue until it works.


----------



## occg.hydro (May 22, 2009)

I wouldn't bother with tissue culture / micropropagation. It's not really a cost effective option. It also requires a sterile work environment like a HEPA flow hood or a glove box. Not that it can't be done, in fact there's a really good series of videos on youtube explaining the process. There are also several scientific research articles out there that list the needed hormones for shoot generation and rooting as well as showing side by side results from different hormone combinations. As a botany student I wouldn't recommend jumping in to tissue culture until you've got your problems with vegetative propagation worked out. It's a much simpler process and the costs involved are much lower, making it less of a loss if you mess up. Tissue culture is going to require a SIGNIFICANTLY larger investment to get up and running and is going to have a MUCH higher failure rate outside of a lab setting than traditional vegetative propagation will. Although the potential exists to exponentially reproduce a mother plant in a short amount of time, not enough research has been done on micropropagation of cannabis. It's much easier to do with african violets or certain species of orchids as there have been lots of lab trials to determine what the best combination of hormones and environmental factors is. If you have money to blow and you're up for a SERIOUS challenge (or you somehow have access to your own personal lab) give it a shot! Either way, I'd suggest going to youtube and looking up "micropropagation" or "plant tissue culture", there's a lot of really interesting stuff on there! If you have any questions about the process feel free to PM me and I'll try to help you out.


----------



## occg.hydro (May 23, 2009)

ozman said:
			
		

> I keep my ph in the cloners,bubble or aero,at 5.5-5.8.
> Ive tried scarification,splitting the stems, all the dips and gels and powders and liquids you could just about mention.



Try upping the pH to 6.0 & see if that helps. I'd also recommend anti-wilt spray, it's good stuff. Got a couple questions about your cloning setup. What's the water temp in the bubbler? Do you use a heat mat & humidity dome when you're using a seedling tray & if so is the heat mat on a thermostat? What's the ambient temp & humidity in the room/closet/cabinet/wherever you're doing this? What kind of lighting are you using? When your clones fail do they shrivel up and die before or after rooting or do they just go on and on and on without growing roots?

The gel on your dying cuttings is either fungus or bacteria, probably both. Either way it's not good. I'm assuming that was in the bubble cloner right? Did you try using hydrogen peroxide with the water and did you make sure to wash the rubbermaid tub with bleach beforehand?


----------



## ozman (May 23, 2009)

Thanx for the interest in my problem lol,
My last attempt was in a aerocloner as per the diy in the forum here,I placed the cloner in my living room,I kept the temps in the res between74 -78,ph 5.5-5.8,changed water every 4 days,I even placed a tote over the cloner for a humidity dome,I kept the lights cfl's equal to 200 watts about 12 inches over the cloner running for 18 on and 6 off,I also used dip n grow in the last batch,thats when I noticed the gelatinous gel on the stems.
I cleaned the whole setup after it was running with a bleach solution and flushed with flora clean then plain water.
I am buying RO water which is 5.1-5.5 ph,with 11-15 ppm.


----------



## smokingjoe (Aug 23, 2009)

With respect OZman, if you aren't able to clone you have buckleys chance of pulling of a micropropagation system, however if you have time patience and money you could start some cannabis pioneering.  

I do intend to install a clean room on my next home, which is probably 12-18months away, and am looking at tissue culture for both rare orchids and cannabis.

Tissue culture is an awesome method of strain hording.  

Interesting you normally don't have to look to far to find a facility that you could tour or volunteer at to see how it's done, it's very common in the nursery and commercial plant industries.

I personally find it very interesting and will share my progress when the time comes.

If you have no objectioin to using Superthrive adding it at 1ml per litre with about 20ml per litre of Seasol should solve your challenge.  

If this doesn't work give it away.


----------



## Tater (Aug 23, 2009)

How often do you change your water, sounds like pathogens are killing your ladies.  Cloning is easy, the only variable here seems to be the person doing the cloning lol.  Keep everything clean and moist NOT WET.  

Cloning in rock wool:  soak cubes in RO or distilled water, take 3-4 inch cutting, scrape sides, stick in rooting hormone, stick in rock wool cube.  2 days under a humidity dome then take it off and let them do their thing.

Cloning in soil: same as above but instead of soaking the soil simply moisten it, don't drown it.

Cloning in water: Change your water daily, pH doesn't really matter as long as it isn't so acidic or basic it will kill your plants.  pH only affect nutrient uptake and there are no nutrients to uptake at this stage.  Chlorine and chloramine will kill off the nodules as they form so becareful.  Don't let the roots sits in water when using a bubble cloner just above the water line, this will speed the process.  Don't put cold water in your tub, or hot water for that matter, room temp.  Keep the temp below 20 C and above 15C thats my money zone.  It will also stop/slow bacterial growth.

Cloning in air: split a branch in half vertically where you want the clone to form roots.  Wrap a hunk of moist soil or rockwool around and between the split.  Wrap it in cellophane or tinfoil, keep moist.  Roots should for in a week or two.  When roots form, remove cutting.

I clone in soil and rockwool, without rooting gel, or doing a second cut.  All i do is basically cut them off with rusty scissors and jam them in rockwool then go do something for two weeks.  I got about 98 percent success with the rusty scissor method lol.


----------



## Tater (Aug 23, 2009)

> ,changed water every 4 days,



Sorry just read that part.  Try changing it every other day or even daily if you can.  Keep the temp the same between water changes ie: do not put 8C water into your tub after removing 20C water, this will shock and kill your babies or at least severely retard their growth.


----------



## smokingjoe (Aug 26, 2009)

Hey B,

It would appear your cloning issue would be entirely environmental.  

Perhaps you are trying too hard.

Do you have a problem with Superthrive, if not try my recipe above.

I will also use a pressure cooker for an autoclave; although used good quality equipment isn't that hard to come by.

I was thinking of adapting a stainless range hood for the laminar flow hood but might just spring for a glove box inside the clean room.  What I find particularly attractive is the ability to maintain the plantlets invitro almost indefinitely.  

Although I'm sure it will require significant trial and error (lots of the later) the theory is not that difficult to comprehend. 

At this point I'm just working out exactly what I'll need.


----------



## occg.hydro (Sep 1, 2009)

hey all! Just wanted to drop by and let you know that BG Hydro is selling a plant tissue culture kit now. Supposedly it comes with "everything you need to get started" including a DVD tutorial. I'm guessing that the hormones included and the references they make to specific amounts would probably be for orchids or african violets (as those seem to be common plants to start with in micro prop). Doesn't look like it includes a flow hood or glove box but you can make one fairly easily. If anyone gives this a shot let me know how it works!

hXXp://www.bghydro.com/BGH/itemdesc.asp?ic=PRAPTCCK


----------

