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"Synthetic" Cannabis Seeds - Explained

dank.frank

dank.frank

ef.yu.se.ka.e.em
ICMag Donor
Veteran
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3550375/

The full .pdf can be downloaded.

Abstract:

Axillary buds of Cannabis sativa isolated from aseptic multiple shoot cultures were successfully encapsulated in calcium alginate beads. The best gel complexation was achieved using 5 % sodium alginate with 50 mM CaCl2.2H2O. Regrowth and conversion after encapsulation was evaluated both under in vitro and in vivo conditions on different planting substrates. The addition of antimicrobial substance — Plant Preservative Mixture (PPM) had a positive effect on overall plantlet development. Encapsulated explants exhibited the best regrowth and conversion frequency on Murashige and Skoog medium supplemented with thidiazuron (TDZ 0.5 μM) and PPM (0.075 %) under in vitro conditions. Under in vivo conditions, 100% conversion of encapsulated explants was obtained on 1:1 potting mix- fertilome with coco natural growth medium, moistened with full strength MS medium without TDZ, supplemented with 3 % sucrose and 0.5 % PPM.

Encapsulation Matrix:

Sodium alginate was added in the range of 2-6 % (w/v) to full strength Murashigeand Skoog’s medium (MS) with or without 3 % sucrose. The solutions were supplemented with 0.5 μM Thidiazuron (TDZ) and 2.5 μM indole-3-butyric acid(IBA). A broad spectrum fungicide, Plant preservative mixture (PPM) in a range of 0.3-0.5 % was added to the gel matrix for in vivo experiments. For complexation, different concentrations (25-100 mM) of complexing agent (CaCl2.2H2O) were prepared in liquid MS medium containing the same adjuvents as the sodium alginate matrix but excluding the PPM. Both the solutions were autoclaved separately for 15 min at a pressure of 1.1 kg cm-2 and temperature of 121 ºC after adjusting the pH to 5.8.

Formation of beads:

The beads were formed by dropping explants mixed with sodium alginate solution into CaCl2.2H2O in a flask, placed on an orbital shaker at 80 rpm. The resulting beads (0.5-0.8 cm in diameter) containing the entrapped nodal segments were left in the calcium chloride solution for 30 min for complexation. These were retrieved using a nylon mesh and the traces of calcium chloride was removed by washing with sterilized distilled water.
Click to expand...


This belongs here. Now we know how. :tiphat:

Image presented for visual of concept only.
14413bioline_08pic1.jpg




dank.Frank
 
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Agronomist

Agronomist

Active member
"Artificial" seeds?
I think you need to change the subject to "substrates for cannabis tissue culture".
Not seeds in any way. This method of propagation is the exact opposite of sexual propagation. It is asexual all the way.
 
M

Mengsk

Active member
Thanks Frank. This sounds like creating little pellets from tissue culture which can be planted like seeds. It sounds very cool but doesn't look like something that can be done in an afternoon with $15 in supplies from the grocery store. I'm curious to try it at some point or see who else has had success.
 
dank.frank

dank.frank

ef.yu.se.ka.e.em
ICMag Donor
Veteran
@Agronomist - Synthetic seeds is the proper terminology, technically. It's a self contained, aseptic - pellet - that holds a viable tissue sample, that can be "germinated" into traditional media, and grown to maturity. Thus the comparison to a "seed". I didn't create the terminology, I'm just relaying the information.

@Mengsk - Precisely. It takes some work, for sure, but Grey Wolf has done it since about 2012. I see plenty of practical uses for it. I think more and more, people are venturing into laboratory type work with cannabis to improve their gardens. It's inevitable. Cannabis growers have always been underground scientists, the available information is just expanding, and with that, so is the skill set, and perhaps the specialties of various growers and breeders.



dank.Frank
 
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dank.frank

dank.frank

ef.yu.se.ka.e.em
ICMag Donor
Veteran
This study only showed they could be germinated in a soil media and grown to full maturity after a 6 month stasis, I did not see if it gave a time limit though. It was also only successful when tissue samples were taken from a plant already established in culture. All the same, it holds a lot of interesting potential for planting large scale clones from "seed". Imagine getting 10,000 synthetic seed of the hemp line you need to mass produce CBD safely without breaking the THC threshold. That would be a lot quicker and much more affordable to the farmer than having to keep a couple greenhouses all winter for mothers and then managing a clone army of 10,000.

I posted this because it is something that has been discussed heavily at different times by many people, Chimera included. It's always been sort of hush hush and a lot of gaps left in between a few hints here and there that it could be done.

I just wanted to share the information for those interested.



dank.Frank
 
M

Mengsk

Active member
Gray Wolf knows quite a bit then :) I've been looking into distillate/vape pens more recently and the processing equipment looks rather expensive. $5-10k minimum perhaps for the necessary solvent recovery and purge stuff. However Gray Wolf built and described about every low cost solution one could think of. Wrote the book on the subject it would seem. Unless working in a lab or working with investors (or investing significantly) a professional/industrial setup is quite a buy-in. This is before knowing how the process will pan out in the long run. In the current market I am not able to afford my supply retail let alone distillate cartridges, and also they are often weak and less than what I was expecting, exacerbating the high cost issue. It's twofold they are way too expensive to buy and they are also not very strong objectively speaking. That's probably a product where you feel the markup the most, some might say price gouge or ripoff. I'm not sure I'd go that far but it's $35 up to ~$80+ for less than 1cc of what is essentially vegetable extract (as in essential oil, lavender oil, citrus oil, almond extract, vanilla extract). The processing cost has a high barrier to entry preventing many growers from experimenting with or producing distillates. To be fair I'm not really sure of the difference between any of those extracts other than solvent vs non-solvent (dry sift or water hash).
 
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Chunkypigs

Chunkypigs

passing the gas
Veteran
It's an intriguing tek but ten years now since the study and you would think if it was practical someone would be doing it.

If it happens that it is affordable I don't think we'll see it anytime soon on the black market but DHN was selling their tissue cultured clones for just $2 more than a normal clone pre legalization.

I'm thinking the clone market in Cali must have shifted to black market now that you can only buy 6 a day from licensed shops instead of 99 like before.

Kevin Jodrey has a new tissue culture lab that looks like it might go up against DHN in the "lablet" clone market, so far he said that the lab has been able to clean up viruses from infected genetics.

dank.frank said:
This study only showed they could be germinated in a soil media and grown to full maturity after a 6 month stasis, I did not see if it gave a time limit though. It was also only successful when tissue samples were taken from a plant already established in culture. All the same, it holds a lot of interesting potential for planting large scale clones from "seed". Imagine getting 10,000 synthetic seed of the hemp line you need to mass produce CBD safely without breaking the THC threshold. That would be a lot quicker and much more affordable to the farmer than having to keep a couple greenhouses all winter for mothers and then managing a clone army of 10,000.......
dank.Frank
Click to expand...

Have you seen the hemp seeds that socio is making, some are 300 to 1 CBD to THC so the BHO from some strains pass US hemp regs. Check his thread.

Seems like clones might not be used much for CBD except maybe for some boutique stuff.
INdustrial hemp in Oregon
https://www.icmag.com/ic/showthread.php?t=330712
 
Chimera

Chimera

Genetic Resource Management
Veteran
Very cool old paper from 2009

Thanks for posting it here on IC again Frank!

picture.php
 
MJPassion

MJPassion

Observer
ICMag Donor
Veteran
Thanks for posting this tech DF. (again apparently... hehehe)


This could make hemp farming for CBD a lot easier, ensuring that each seed is an exact replica of the mother genetic and making a very uniform crop.


Only downside would be if that particular crop variety came under fire of dis-ease, the whole of the crop would be at risk.
 
Chimera

Chimera

Genetic Resource Management
Veteran
Zero chance of that happening MJP. When you plant acres, you plant from seed - even cuttings are too inefficient for field crops - which is essentially farming for single compounds, not flowers.

These propagules are inefficient for mass plantings, even for planting small crops- they are labour intensive to make, and require babying to bring back to animation.

Their worth as we investigated them is as a method to transfer clonal material discretely. More recent research has identified Thidiazuron (TDZ), the PGR used in this paper, as a major source of induced mutation in the tissue culture process, causing upwards of 250 base-pair substitutions (mutations) per cycle of tissue culture.

These structures are also referred to as uni-polar - meaning they have a vegetative growth meristem, but no root meristem - so they need rooting like clones do. There are other methods like somatic embryogenesis which solve this problem, and somatic embryos can also be encapsulated a la artificial seed.

-Chimera
 
unnamedmike

unnamedmike

Well-known member
Chimera said:
Zero chance of that happening MJP. When you plant acres, you plant from seed - even cuttings are too inefficient for field crops - which is essentially farming for single compounds, not flowers.

These propagules are inefficient for mass plantings, even for planting small crops- they are labour intensive to make, and require babying to bring back to animation.

Their worth as we investigated them is as a method to transfer clonal material discretely. More recent research has identified Thidiazuron (TDZ), the PGR used in this paper, as a major source of induced mutation in the tissue culture process, causing upwards of 250 base-pair substitutions (mutations) per cycle of tissue culture.

These structures are also referred to as uni-polar - meaning they have a vegetative growth meristem, but no root meristem - so they need rooting like clones do. There are other methods like somatic embryogenesis which solve this problem, and somatic embryos can also be encapsulated a la artificial seed.

-Chimera
Click to expand...


thanks chimera about tdz and mutations:tiphat: can you share the paper please ?





I believe the future for large crops, including hemp, is to make self-replicating plants, self-fertilizing organisms that pass an exact copy of their genes to their offspring. As an example you can search "Mutant Crayfish Clones Itself" in a search engine to see a natural case of what I speak. I think Crispr is the tool for it.
 
C

clearheaded

Active member
ya wonder if segra is going to be using TDZ?, Bman thinks its organic, no PGRs in tissue culture hehe. anyhoo off topic.

ya neat but wont be able to toss in fridge for few years or cheaply send in envelope across the world.. and ya is sorta like caviar similar process for "vegi cavier" or molecular gastronomy. essence of lavender beads in nitrogen soda.

there is clones being used in feild propagation for hemp cbd var. however your right, just need seed stock to be developed which will out produce any gains from having elite cbd hemp cuts. also opens up feilds to disease, talk about mono culture. i do see this happening as regulations prevent farmers to cycle crops cannabis>legumes>flax>cannabis..or extensive chemicals be used as it is a more lucrative crop. both are bad. if strip biomass of cannabis feild for a decade cannabis being a pretty good at sucking up micros and macros will strip the soil pretty well i would assume. anyway off topic. cheers!
 
Sunshineinabag

Sunshineinabag

Active member
May I ask u guys who have LEGITIMATE experiance in breeding cannabis!?
What goes into making a new cultivar variety?
In 2014 I cleaned out a college dorm that had Nike shoebox full of seeds. A lot were not viable and were flat and yellowed out. Turns out the kid had a relative that shipped him a half elbow throughout his years at university of Vermont and anytime he found seeds he saved them, mind u this was like 2008, so I was part of another site back then and I sent two guys one in northern New Jersey and the other in Virginia like 85 seeds each.........a year later this one guy all of the sudden claims to have his own varietal he created!? So he got seeds from me, then blows smoke up folks asses telling them he created a new cultivar variety............I thought he was full of chit.
 
romanoweed

romanoweed

Well-known member
They are really cool for mailing genetics, but in cold storage they are only storable for a Year.

They also requiere the reciever to grow it up in Tissu culture.. It doesent sprout in the Soil.. Those are the minus points.. But i wished Clonesharing was more common, and this would be a neat solution .. And i heard its not very complicated for the grower to revive it..
 
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