Micropropagation - Plant Tissue Culture

Omg these grow boxes, pressure cookers, sanitation and small jars reminds me of the days when some people grew shroomz like crazy idk who but just some people haha :D GL hope it works out for u.:thumb:

my back ground is in mycology :rofl:

:hippy:
 
I got a secret love for mushies too. Looks like you trying to get this down to breed some kind of super herb I hope it works out for u. Just don't go all gmo on us :D if you get a sexy plant from it. GL :goodjob:
 
I've been curious about tissue culture for awhile now, and your thread is really fascinating. I do think it will be the way of the future, as weed comes out of the basement and into the well-funded lab. I look forward to giving it a try some day--no time now for another project though.
 
I got a secret love for mushies too. Looks like you trying to get this down to breed some kind of super herb I hope it works out for u. Just don't go all gmo on us :D if you get a sexy plant from it. GL :goodjob:

thanks! :love: i'd rather assist nature's plan and give it opportunities than try to force her :) no GMO here to be sure.


I've been curious about tissue culture for awhile now, and your thread is really fascinating. I do think it will be the way of the future, as weed comes out of the basement and into the well-funded lab. I look forward to giving it a try some day--no time now for another project though.

once the big corpora' gets done.. it will eventually trickle down to us lowly patients and personal growers. like cell phones, computers etc. our grand children's grand children will probably be growing from gel caps in space not knowing what a real seed is lol.

i did some mycology stuff the other day with no troubles so this might not be as bad a spring as i was expecting. once i get the clones out of the glove box again i will give this another go. it's been on my mind a lot lately :)

thanks for stopping by! :thanks:

:Namaste:
 
Home now, smokin' Cenex and the Kratom is kicking in. Almost good to go lol.

Here's an example off an easy home made Still Air Box (SAB).
SAB.jpg

I'm not going to post the directions simply because there are so many variations. This can even be a cardboard box with clear plastic lol. All kinds of corners can be cut when designing this. I was actually thinking of making one real quick using a fish tank on its side with plastic sheeting and duct tape. I have a plastic tub but I would like to take pics while working possibly. So the glass would be nice.
Anyways, the objective is to create a mostly sealed environment that can be cleaned with bleach or alcohol spray. Disinfectants like Lysol can be used, but it has a history of causing a cellular disorder in fungi known as rosecomb. I have no idea how it will affect plant tissue and we are not even going there lol. I'll make the box tomorrow and then we can do some culture work.
Normally I would break out the big gun for this sort of work, but I want to show everyone just how easy home culturing can be when you get used to it.
This is a back pressured clean room with 2'x4' Hepa filter laminar flow hood. Not your everyday home appliance :laughtwo:
And not something I get to show off very often lol. So I had to sneak it in here somehow. :icon_roll
Honestly, though a bit cumbersome, an SAB can be every bit as effective.
clean_room_1.JPG
clean_room_2.jpg


Much love!
:Namaste:

I'm referring to you as Heisenberg from now on lol
 
I bought some seed tissue culture on ebay. The seeds have been init over a month and nothing is happening. Maybe I'll make my own. Just need some more seeds first.
 
Well, it's good that you got it then. :laughtwo:
 
Still going ahead with it all, love an update mate

I haven't forgot :)

I've been able to sneak in a little reading from the book here and there. Things just got out of control this Summer lol.
However, I do need to start jumping on some mycology projects for Fall so I'll get in there to clean the clean room and fire up the flow hood. It will make things a lot easier even if it's cheating.

Thanks for bumping this! :high-five:
 
All good brother I look forward to seeing your work also add a link to this j in my j in my sig, get somecmore numbers in here to witness the wonder
 
I've made two adjustments to the approach. These two items cost about $20 US. The larger bottle is a product called Tween. It's basically a cleaning solution for the plant material we had problems with earlier on. The material spends about 15 minutes soaking in this before going onto culture medium. We can kiss our little brown mold any other lurkers deep in the plant tissue goodbye. :)

The smaller bottle is the culture medium itself. From what I've learned Cannabis is extremely picky. Not so much in the fact that you can't get cell division on various mediums. Which we did. It's getting good cell division. There seems to be several issues that I'll get deeper into later on.

This "MS Medium" is proven for Cannabis :thumb:
(Murashige and Skoog medium (or MSO or MS0 (MS-zero)) is a plant growth medium used in the laboratories for cultivation of plant cell culture. MSO was invented by plant scientists Toshio Murashige and Folke K. Skoog in 1962 during Murashige's search for a new plant growth regulator.)
Micro-Prop_medium.jpg

We'll still be working with the sugars (adjusting to 30 grams per liter) and hormones. There's one more ingredient that I'm still researching that is a little bit of a concern for later stages. It's expensive so I'm trying to see of there is a way around it lol. I've got a busy day ahead of me, but I'll try and at least get a batch of culture made today. No promises :laughtwo:

Thanks again for your interest and ultimate patience :Namaste::circle-of-love:
 
I wasn't able to get a good pic of the yeast, but if you google yeast on agar there are plenty of good pics. I did manage to get a picture of another hitch hiker. This one also started at the cultured tissue and grew outward. An indicator that it was present in the tissue like the yeast. It's a species of Fusarium. Not sure which one.. but it is a soil/plant dweller.

Fusarium is a large genus of filamentous fungi, part of a group often referred to as hyphomycetes, widely distributed in soil and associated with plants. Most species are harmless saprobes, and are relatively abundant members of the soil microbial community. Some species produce mycotoxins in cereal crops that can affect human and animal health if they enter the food chain. The main toxins produced by these Fusarium species are fumonisins and trichothecenes.

The name of Fusarium comes from Latin fusus, meaning a spindle.

Fusarium.JPG

I don't think you want this stuff close to your gardens... That word fusarium and fugus are scary words to me. Sorry I don't do alot of reading myself,, but the fusarium fungus sounds WAY too close to wilt. That if it is that one,, and I think it is, it's a real nasty''' soil borne spore,,, that attack the plants vascular system, causing the plant to,'wilt and die eventually'' stuff is hardy, it winters over easily, in like Northern Ill and other cold as hell places, with lots of snow. But it survives them winters and reinfects the soil soon as temps hit the target temps ans R/H for it,,, and starts all over again. You see a pretty patch of say 50 plants,, and one here and one there are all wilted up and turning brown,,, it's that shit... Just opening my big mouth but ya'll probably said that on the prior page..

I was wanting to know if you have been able to get anywhere doing the Tissue Cultures? Pretty involed process. But if one was wanting to do some serious tests, what better way to get the subjects. Hope you don't mind I come take a peek every now and then,,, Hopefully you be screaming,,, IT"S ALIVE"
 
I had a few set backs :( One of my medical conditions decided to rear it's ugly head. Got very sick and when I went back to work I found myself in another position which I have been told is temporary due to the loss of a manager. Better be. It's a time leach job hmmf. At any rate there is some positive in my negativity :laughtwo: I'm scheduled for a surgery on March 3rd. I'm not real thrilled about it lol, but it will finally allow some time to get back to this. Otherwise I have everything I need. :)

Yeah some Fusarium can be nasty little things and definitely associated to stem and bud rot. This one in particular seems fairly benign. I'll be treating the next tissue samples in a 15 minute dip of Tween. That should rid us of any unwanted hitch hikers. Contamination after that will be all my fault lol.

Really good to see you here Norcal' :high-five:
 
Tada! :cheer: Making another batch in a few and just making sure I had all my notes. I won't be able to be as detailed initially due to being spread too thin as usual, but this is for a business project now so no playing around. I'm skipping the box and using the flow hood when the time comes. Right now I'm busy cooking a media recipe that is proven to work.
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:Namaste:
 
Did they grow?

Same thing again. They get started and then the little invaders come. They are definitely coming out of the plant tissue so for the next try I am going to severely increase the sterilization time. The directions for the Tween is 15 minutes which I was a little suspect of. Seems short for such a soft solvent. Otherwise everything else seems solid. We'll get a clean sample eventually.

:Namaste:
 
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