Review on Cannabis Tissue Culture

[anon64373531]

For your continued edification:

REVIEW
published: 03 March 2021 doi: 10.3389/fpls.2021.627240

Medical Cannabis and Industrial Hemp Tissue Culture: Present Status and Future Potential

https://fjfsdata01prod.blob.core.windows.net/articles/files/627240/pubmed-zip/.versions/2/.package-entries/fpls-12-627240-r1/fpls-12-627240.pdf?sv=2018-03-28&sr=b&sig=X33RI1bdINmvyR8hxTVQmk2Fg%2FdHVmC1zri9Q5ktNjU%3D&se=2022-08-20T21%3A01%3A29Z&sp=r&rscd=attachment%3B%20filename*%3DUTF-8''fpls-12-627240.pdf

[Ralf]

Thanks; this is a new paper for me. There are a quite a few TC and micropropagation papers on cannabis now, and I can’t wait to see what it comes out in the next couple of years.

It’s incredible to see the rate of new papers on cannabis cultivation, breeding, IPM, extraction, processing, TC, and genomics compared to even five years ago. And the quality keeps improving, too.

The link you shared is broken in my browser (maybe from my security add-ons), so sharing an alt link here: https://www.frontiersin.org/articles/10.3389/fpls.2021.627240/full

[Ralf]

From Could anyone help me hunt down Starburst OG? :

@anon64373531

Holy shit. $500 per culture? That gets you how many explants?

Tissue culture is on my bucket list of skills and it just jumped higher. Especially synthetic seed tech.

Same here. And in the future we may not have to mess with synthetic seeds as a means of long-term cryopreservation:

Cryopreservation of Shoot Tips of Elite Cultivars of Cannabis sativa L. by...

Also, this is a good paper in relation to maintaining long-term genetic fidelity though TC regeneration:

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Viruses, Viroids, and Cannabis: A Brief Primer

@Ralf

TC can take care of viruses and viroids. If someone has cuts they love, TC can do wonders for its fidelity, too, if you can find a TC lab that does DNA demethylation.

Thwarting a Potential Cannabis Catastrophe With Science

@FicklePickle

At around 3000 a cut. And then it’s still not guaranteed to work

@Ralf

Yeah, def, not cheap. And takes a long time. I haven’t contacted Front Range, but if they’re at $3,000, I hope they would offer a partial refund if they didn’t succeed. I’m looking for labs in the US that offer this service to price compare. There def is a market for this, esp at a lower price point, so I hope to see more labs in the future.

@BananaBannabis

We use someone that does node culture for around $500

Meristem TC is more expensive and takes longer. It is my understanding meristem regen should “clean out more” then typical node culture. However node culture still has a high probability of cleaning out contaminants.

@emdub27

Front Range Bio can’t even keep their own stock clean. Specifically at the Salinas CA location. Stay far away.

[Ralf]

A couple of posts from Haze And Sour seem on topic as well:

Haze And Sour

To take the Sour D project one step further, I have this project flow in mind: Identification of the ‘real’ Sour D cuts using whole genome sequencing based on comparison to Chemdog 91, AJ’s Sour D, and agreed upon Sour D trait identification @silverstudent suggested Detection of DNA methylation through tissue culture of the selected Sour D cuts and the Chemdawg 91 cut. (This step may be unnecessary because it’s probably safe to assume the ~27-30-year-old genetic age of the cuts has introdu…

Haze And Sour

Maybe. With enough depth of coverage you could probably work out how many individual mutations you were looking at. When I was last worried about somoclonal variation (I’m of the camp that doesn’t draw a line between tissue culture induced vs “natural” genomic/phenotypic changes) we didn’t have access to massive sequencing…so I have to admit I haven’t seen data on exactly the kinds/scope/range of mutations or their frequency. I could make the same guesses I would have 20 years ago, re: deleti…

Haze And Sour

Thanks for the info! This is a topic where I know what I don’t know is vastly greater than what I do know. So, I love that you’re on this site and willing to share your expertise I have to stop by and buy you a beer or twenty the next time I’m on the west coast . Is it your opinion that somaclonal variations and somatic mutations are the same (or have the same result) but are from different causes? With the former mainly caused by newly induced mutations from some TC processes, an…

[anon64373531]

I tried to volunteer with Charlotte’s Web about 10 years ago in exchange for learning tissue culture. I thought they’d love to have a PhD chemist as a volunteer. Nope. More interested in protecting their IP.

[Ralf]

Their loss. Are they even a thing anymore? I assumed their 15 minutes of fame ran out.

[anon64373531]

Once CBD isolate hit $300/kg I’m sure they became toast in short order.

[SISU]

Not really - they created a THC brand a year or two ago that’s sold in CO+FL, but I haven’t heard much about it

[standardoil]

So who is gonna be the first smart kid here to start a business and offer complete equipment and supply packages for TC in the weed industry

There are 19 fake terpene companies and at least a dozen that sell you French fry oil powders for crc. Surely someone has got to see the untapped market

[SISU]

@standardoil

So who is gonna be the first smart kid here to start a business and offer complete equipment and supply packages for TC in the weed industry

You down @Cloner ?

[standardoil]

By package I mean down to the wire racks with lights on them. Hood, packaging, tools, sterilization stuff, media and hormone mixes

[FicklePickle]

I think cloner is making plans to get out of all this. At least that’s what we’re hearing

[SISU]

If true I don’t blame him ha, frankly we should all slowly be working on contingency plans

[FicklePickle]

As entrepreneurs it should be in our nature

[Ralf]

@anon64373531

I tried to volunteer with Charlotte’s Web about 10 years ago in exchange for learning tissue culture. I thought they’d love to have a PhD chemist as a volunteer. Nope. More interested in protecting their IP.

Are you near Mass? This looks interesting:

https://www.instagram.com/cannomicslabs/

Cannomic

[Ralf]

Tetraploids anyone?

I have used the seed treatment method with oryzalin, but this method has piqued my interest! Note that polyploid cannabis hasn’t shown benefits except for creating sterile plants at the triploid level. Many people claim it increases yield, THC, etc. However, all but one of the published papers I have read reported no significant increases. The one paper that reported a significant THC increase is an outlier, and the increase isn’t impressive considering the starting THC %:

Development of a Direct in vitro Plant Regeneration Protocol From Cannabis...

[anon64373531]

I am currently in MA, but don’t have $2500 to drop.

I already know asceptic technique and mammalian cell culture, I’m sure I could figure it out.

[Ralf]

Do you feel like moving to MI next year? I’m planning to keep a few dozen strains in rotation at all times, and I want to set up an in-house TC lab at our cultivation facility. I want to regenerate the mother stock by TC approximately every six months and replace the mother stock with cryo-stored tissue every few years.

I would love to hire an industry-experienced Ph.D. chemist and have the company pay for TC training. The work would include running the TC lab, either running or acting as the lead chemist of our processing facility, and running our in-house analytics (GC-FID, HPLC, and hopefully adding MS in the future).

[anon64373531]

Pm’d you.