Tissue culture cloning

Mestizo

Member
Interesting stuff. I want to look into tissue culture cloning for germinating ancient seeds, because I'd like to tease some old landrace Mexican and Colombian genetics out of a batch of very old hippy seed I recently acquired.

Interestingly, for my purposes, the best method may be at the other end of the sterility scale: bio-priming by germinating the old seeds in worm dirt to bathe the seeds in good microorganisms that kill and compete with the bad stuff that otherwise tends to rot old seed embryos.
Hey Marcus. I visited the micropropagation department at Kew Gardens recently and was shown "seed traps" that they use to germinate seeds that are difficult to germinate in their natural surroundings. The specifc example they showed was this Viola which had been dying out in Northern England and Scotland due to the plant's inability to self propagate through rhizomatous material and the difficulty that the plants had fertilising female sexual organs via male pollen. They were creating tens of thousands of propagules to repopulate areas that they had previously been growing in, relatively robustly. Any way, they had identified different strains of mycorr that invade or form associations with the seed and then again different mycorrizhal and bacterial strains that helped seeds along the way once germinated.

I always wondered whether you could generate propagules from leaves in a similar way to propagating Heuchera spp. by making incisions at veinal junctions then pressing firmly in to a low nutrient growing media so that all points of the leaf touch and are firmly bedded down. Would the meristematic tissue develop propagules from leaf lamina cuttings in Cannabis I wonder? I think hygiene would be less important with this method but obvs would still make an impact.
 

sensient

Well-known member
hey guys, In the early 2000,s Breeder Steve of the now dufnct seed company spice of life was going to release special beens. I don't know the full details but basically he was going to select the best phenos of each of his strains and clone only such as sweet pink grapefruit and others that were available at the time, he was goingto perform tissue culture and encase the cultures into small seedlike balls. He didn't go into much explaining about them but said after a month of treating them in a certain way you would basically have a clone,that you would treat as a normal clone and grow out,he didn't explain much but said all details would come with the packs of ball, I will look at the old cannabis world forum and see if I can find the post,as a lot of the old post had been saved and may be available in the spice of life seeds help desk. it was entitled: seeds will soon be obsolete. Obviously he was talking about tissue culture being in these balls. The project never went ahead,i guess he realised it may be not as easy as he thought, but he said the method hd been perfected and anyone would be able to grow out these balls. it seems like a god idear in theory and would make clone onlys available to everybody. I will go have a look for the thread now and post a link if I can find it. it would be great if this was possible. a lot of breeders could make selectionsof there best phenos and release these balls to people who wanted them, and would probly make the S1 seed situation less viable,as instead of s1,s you would have the actual clone available. Same with r1 hybrids, as breeders could select the best phenos of there reversed hybrids and do the same thing. of course I wouldn't be up for companys releasing these only and not giving us a chance to find our own phenos from a pack of seeds,but it would make obtaining clone onlys a hell of a lot easier for those who want them andput this clone only I am ther king god complex idiots to rest, as we would all have access,anyway will go have a look
 

Broseidon

Member
I mean... basically he wanted to sell tissue cultures to people so they have clones instead of seeds and as a gimmick wanted to make them look like seeds.

But nobody would be able to put them in soil and have a plant grow from it as the tissue cultures would have to be awoken from their suspended state first through other reagents/treatments.

So the whole "ball" or "make it look like a seed" part was basically marketing on his part.
It was just a way to encase the tissue culture sample. He could have sent it in any form of box really that allowed the TC to remain sterile and frozen.

Back then, I doubt it was possible to easily send it in a frozen state across the world. Now it might be more possible but still likely expensive.

And none of this deals with the problem that once the TC arrives at its destination (in a "ball" or any other type of container), the recipient has to provide a sterile environment and certain reagents to make it grow.

I mean, in theory, I suppose you could encase the TC in a non-reacting layer, then encase that in the necessary reagents to make the TC start growing and encase that in another protective layer. Then need to get all of that suspended (i.e. frozen).
Then if you could get that in the suspended state to the customer, I suppose it could theoretically be arranged in a way that at room temperature the protective layer between the TC and the active reagents dissolves. If air is not required, you could make the outer shell in a way that provides a sterile environment for the duration of the reaction between reagents and TC and make it so that this outer layer dissolves once the TC has grown enough to be able to handle the soil (i.e. once it has the first roots).


I can imagine it in theory. I mean that is close enough to how seeds work as it is and would just be another instance of imitating nature. But I can't imagine someone like Breeder Steve to have both the aptitude and access to necessary components/tools to realize this.
This is the kind of advanced bio tech that Big AG and Big Pharma are good for. The kind of stuff you need these big bastards for with all their deep pockets and resources.
 
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sensient

Well-known member
hey all,could,t find the post.
hey bro,thanks for the info,makes perfect sense,as I said I haven't looked into the tissue culture tech much myself,but what your saying does make perfect sense.yeah definatly aother gimmick but an intresting one,but by what your saying would not be viable for the average seedbank. to much science involved.would defintly be an issue packaging these things and keeping them viable for postage. good idear in theory, but can definatly see what your saying,cheers for the info, you,ve definatly got some knowledge in your head,great stuff mate. keep it up! learn something new every day,even an old fart like me,cheers..S
 

59lespaul

Member
When I did it back in the 90s, my kit had test tubes with agar on them The tissue, has to be placed Vertically/Buried 1/3 of the way into the agar, not Flat, or horizontally. From what little I know, you cant surround/completely cover the tissue with anything. Nodes start to form on the tissue. I used 1 Sq/Cm of leaf tissue, which also has to be sterilized. I cant remember the complete processes.

But things change, and they can do anything now. So I'm probably really out of date. Probably could send in liquid nitrogen.

Mine was buried 20%/30% thin side of the tissue in the agar, and bumps eventually show up.

Also you cannot at all expose them to even any kind of UVA/UVB. Even a quick flash from a distant window, WILL KILL THE SAMPLE. PERIOD.
The tissue/plantlets are done under cool white, and as they grow transitioned over to a combination Warm/Cool, and then individually separated, and placed in jars with more Agar ect, and grown to several inches, and then put into medium, and UV hardened.
Its not as hard as it sounds, and is best just to do in a place that has no carpet, no curtains, windows, drafts, can be completely sprayed down with Lysol. Lysol is your constant friend.

The main thing is sterile, nothing floating in the air contaminating the sample, and No UV exposure.
 
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sensient

Well-known member
hey all,hey Cannafish,thanks for that,yeah that was a later post I had forgotten about. The post at Cannabisworld was entitled "seeds will be obsolete", he basically claimed he was doing it and was a sure thing and underway. After not hearing more about it he later posted the posts you have shared. Obviously things were not a sure thing at all and he was speaking a little bit of shit.But yeah that's what he called them, artificial seeds. Micro clones ashe puts it. he said they would take a month of treating them with the instcuctions he would provide,then you could treat them as normal clones and grow them out. I am not sure about the Chimera thing but maybe this was the partner involved, I wouldn't know but it very well could be. Soan to those in the know, would this be a viable thing? could these artificial seeds be produced? I am just interested because if this could be done it would definatly change the elite clone world and put these god complex fools to rest, no more 5-25000$ prices some of these idiots try to charge for so called elite clone onlys! I'm not that interested in the clone onlys,but a few of them I would like to obtain. Mainly the original 91 chem. theres a lot of opinions regarding the clone,but it definatly is something special and I would not mind trying it out for myself.anyway, I'm getting off track from the op discussion. Just thought these artificial seeds sounded interesting, take care guys..Sensient
 

Broseidon

Member
It's definitely interesting, would definitely be a game changer and at least in theory is definitely doable.

But I would bet my left testi that IF this ever becomes commercially viable (i.e. feasible, i.e. it's not so expensive to do that you have to charge prices that customers won't pay), it will likely be accompanied with all kinds of trademarks by Big AG and Big Pharma...

IF this ever becomes a commercially viable solution/procedure, it will not be something us small timers or the cannabis community as a whole will benefit from. It will be something that Big AG and Big Pharma benefit from.

Because they are the only ones who could realize something like this (imho) and they sure as shit aren't known for doing things out of the goodness of their heart or because it is the right thing to do or because they want to advance the cannabis community or whatever.
IF they will realize this, they will do it for the money and power/control it affords them and then we won't want to have it, I can all but guarantee you that.

Maybe with ongoing legalization and advancements in technology, some small timer like one of us will be able to realize this or a similar tech in the next 10 to 20 years or so. That is the only way I could see this kind of thing going anywhere positive.
 

Glow

Member
Actually information on tissue culture cloning is widely available in the agricultural sector. It's not as if cannabis is unique, the same principles apply re meristem culture. The cannabis industry is years behind other ag sectors with this. However, having spent 8 months working in tissue culture and virus indexing I'd recommend taking a course on it.
 

Glow

Member
Hi All

there is several companies already doing tissue culture for cultivators.

Segra International | An emerging specialist Research and Development Company focused on plant-based medicines, including medicinal cannabis.

this group is out of Canada but there are groups all over trying to do it with Cannabis in large quantities as it is a great alternative for cloning in quantity...well that is one of the main applications.

Glad to see your innovative ways of diy tissue culture...but sterile environment is the most important factor or else it is a mess.

The disadvantage is it takes quite some time to grow out...would love to see a photo of a home system if anyone wishes to show theirs....

all the best Sb
SB yes there are several labs that I know of in the US doing cannabis tissue culture. One in WA, one in the Bay Area, one in Colorado Springs etc.
 

Deach69

Active member
Actually information on tissue culture cloning is widely available in the agricultural sector. It's not as if cannabis is unique, the same principles apply re meristem culture. The cannabis industry is years behind other ag sectors with this. However, having spent 8 months working in tissue culture and virus indexing I'd recommend taking a course on it.
I'm looking into it, thanks Glow :)
 

Glow

Member
Interesting video here
I think it's important to point out that tissue culture is relatively straight forward but won't clean viruses out of the source material. That's far more complex and requires meristem tissue culture along with hot and/or cold treatment and/or antiviral treatment with agents such as ribavirin.
 

shantibaba

Breeder and moderator
Hi All

Well Breeder Steve was in Lugano, Swiss doing it but while it was a good idea the methodology was still going through trial and error. Chimera also well known to me for many years was giving it a go...the trouble with all applications in science is converting it to practical efficiency in the real world. Sterility is by far the most determining feature of tissue culture and long term preservation of hybrids and varietals and experimental mothers and fathers is definitely a purpose fit for consideration of tissue culture....but it takes over 9 months to regrow to useable clone which does not make it economic feature for this work unless done to pre orders a year in advance. I have tried it and followed it in NL and CH for some time but there is yet to be better applications for it than what I already mentioned. It is a great practical way of genetic selection storage for long term...imo. All the best sb
 

Glow

Member
Hi All

Well Breeder Steve was in Lugano, Swiss doing it but while it was a good idea the methodology was still going through trial and error. Chimera also well known to me for many years was giving it a go...the trouble with all applications in science is converting it to practical efficiency in the real world. Sterility is by far the most determining feature of tissue culture and long term preservation of hybrids and varietals and experimental mothers and fathers is definitely a purpose fit for consideration of tissue culture....but it takes over 9 months to regrow to useable clone which does not make it economic feature for this work unless done to pre orders a year in advance. I have tried it and followed it in NL and CH for some time but there is yet to be better applications for it than what I already mentioned. It is a great practical way of genetic selection storage for long term...imo. All the best sb
Yep great way to preserve genetics in terms of storing tissue cryogenically in liquid nitrogen. Also meristem tissue culture along with antiviral treatments is going to be incredibly important in cleaning up the viroid and virus issues the cannabis industry is currently facing. In terms of a way to ship though hmmmmmm.... Prob not from a commercial perspective.
 
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