Cannabis tissue culture

[OldSSSCGuy]

Sorry for joining get mo's bump up, but there is so much brain power floating in this thread that it seems a feasible place I might be able to ask a couple of (what are probably) really stupid questions. Hell, I cannot even pronounce most the the botanical terms used in this thread.

1. Does TC offer -any- potential for regenerating cannabis from old seed? Or is TC a case where its basically micro-cloning living plant material? How does that relate to mentions of DNA techniques?

2. How will/would they regenerate the thousands of plants stored in the Svalbard Global Seed Vault? Are they not mainly stored as seed?

Anyone willing to hold my hand on this one?

 

[Agronomist]

1. Does TC offer -any- potential for regenerating cannabis from old seed? Or is TC a case where its basically micro-cloning living plant material? How does that relate to mentions of DNA techniques?

2. How will/would they regenerate the thousands of plants stored in the Svalbard Global Seed Vault? Are they not mainly stored as seed?

1. Yes.
2. Like regular seeds, They plant them and the plants grow.

 

[OldSSSCGuy]

1. Yes.
2. Like regular seeds, They plant them and the plants grow.

1. Yes that TC can be used to culture old seed, or yes it can only be done with living plant material?


2. Why is there an assumption that all seed stored there will simply germinate normally - give the age and storage technique?

 

[hellfire]

1. Yes that TC can be used to culture old seed, or yes it can only be done with living plant material?

To my cursory knowledge it is only live plant material and seeds that will germinate under certain conditions. I have not seen evidence for cannabis seeds that are essentially dead or so old they won't crack developing into full plants through TC. I would love to see something if this is a reality.

 

[OldSSSCGuy]

I appreciate the opinions. Still sitting on a large stock or original SSSC seed from the 1980's-90's and continue to hold hope that science & lab work might catch up with my seed hoarding.

 

[sbeanonnamellow]

Table 13.1 from Cannabis Sativa L.- Botany and Biotechnology lists the following as in vitro protpcals developed for Cannabis sativa L.:

Seedling parts, root, hypocotyl, leaves of seedling, male and female floral parts, embryo, leaf, stem, anthers, internodes, cotyledon, stem and leaf segment from seedling, seedling, stem, apical and axillary buds, leaf callus, callus, petioles, stem root, lateral buds from seedling, nodal segments with axillary buds, epicotyl, and shoot tips.

*with exclusions for redundancy*

If you'd like the titles of the iterature associated with any of the above let me know and I'll look them up pass them along. Much love

 

[OldSSSCGuy]

Really appreciate the effort, but you are offering concise botanical information which is wasted on me completely and entirely.

 

[Bio boy]

I had aone wifi seeds fAiling to germ n grow well they used to grow awsome now get 1cm and fail.

Could tc be a savior to the gene like this would it refresh the plant so to speak

 

[LostTribe]

To my cursory knowledge it is only live plant material and seeds that will germinate under certain conditions. I have not seen evidence for cannabis seeds that are essentially dead or so old they won't crack developing into full plants through TC. I would love to see something if this is a reality.

I have found no studies or reports of successful TC involving the use of seed much less seed which has a 0% germination rate.

However, one of you should invest in a lab and tech to make it a reality!

You could make or buy a seed cracker, currently sold out, and try the seed cleaning regimen along with the seed cooker deal that uses sponges and floats in a Rez with aquarium heaters...Also people have started worm farms and planted directly into the worm farms.

I tried GA3 combined with sterilizing the seeds with H202 and going the old paper towel route with no success.

 

[Azaghal]

Hello everyone,

for those who are interested, the Ontario Agriculture College (OAC) at the University of Guelph is going to hold a micropropagation workshop from Friday July 14 at noon, until 5:00pm on Sunday July 16.
The cost of this workshop is $3000 (CAD) per student.

These are the links to register: 1 & 2 .

Cheers

 

[Loc Dog]

Can anyone point me in the right direction to first learn how to recover a rare clone that is only living roots with a few inches of chopped stalk, then how to do tissue culture from apical meristem. I have grown shrooms 15 years ago so sterile procedures are not issue but need to know what chemicals to buy and proper mixing and environment. Someone mentioned they would be different procedures but that is all they would say.

 

[DNHappy]

Plant cell technology has great information. I've done tissue culture once and was successful at it for a little while then the plants started dying one me. I still have not got it down all the way but I'm gonna keep trying. I'd love to help you out but all I can say is research. Check out plant cell technology, that's where I learned.

 

[benjuanman]

Plant Cell Technology is good like DNHappy said as well as this sight https://www.researchgate.net/public...issue_Cultures_of_Drug-Type_Cannabis_sativa_L

 

[gmanwho]

Can anyone point me in the right direction to first learn how to recover a rare clone that is only living roots with a few inches of chopped stalk, then how to do tissue culture from apical meristem. I have grown shrooms 15 years ago so sterile procedures are not issue but need to know what chemicals to buy and proper mixing and environment. Someone mentioned they would be different procedures but that is all they would say.

Hey Loc dog, late reply, not sure what you know of that procedure. where you able to get anywhere with reproduction from root? i beleive the procedure is called organogenesis. seems the process is abit more lengthy then apical meristem reproduction.

such an amazing thing to take a root, a piece of leaf, a petiole or piece of stalk. reproduce an or form callus cells, then split off and introduce the different hormones for shoots an roots.

the other day i was floating thru ig an i saw a tc culture that they used a bract. the bract was above the agar, callused, and shooting roots, but not shoots yet. so f'ing awesome.!!!

 

[OldCoolSativa]

I have found no studies or reports of successful TC involving the use of seed much less seed which has a 0% germination rate.

However, one of you should invest in a lab and tech to make it a reality!

You could make or buy a seed cracker, currently sold out, and try the seed cleaning regimen along with the seed cooker deal that uses sponges and floats in a Rez with aquarium heaters...Also people have started worm farms and planted directly into the worm farms.

I tried GA3 combined with sterilizing the seeds with H202 and going the old paper towel route with no success.

I'm sitting on a few grams of bagseed collected from the mid sixties into the early 1980s. After blowing through a few dozen trying various techniques, and only getting a couple to shoot tails, I'm currently sitting on the rest waiting for TC and embryo rescue technology to advance so I can improve my odds. They were gifted to me about 6 years ago and are the treasure of my collection, despite being currently unviable. Operative word there is currently.

It is my opinion that embryo rescue technology requires some micromechanical means of manipulating the seed to help it split open and the safely remove the embryo from the seed husk and from the embryonic (amniotic?) sack. This would all need to be done under temperature-controlled sterile conditions of course, and the process would also require just the right mix of nutrients, enzymes and hormones.

Can anyone point to someone who has demonstrated success with embryo rescue of cannabis seeds that are 40+ years old?

 

[DNHappy]

I'm sitting on a few grams of bagseed collected from the mid sixties into the early 1980s. After blowing through a few dozen trying various techniques, and only getting a couple to shoot tails, I'm currently sitting on the rest waiting for TC and embryo rescue technology to advance so I can improve my odds. They were gifted to me about 6 years ago and are the treasure of my collection, despite being currently unviable. Operative word there is currently.

It is my opinion that embryo rescue technology requires some micromechanical means of manipulating the seed to help it split open and the safely remove the embryo from the seed husk and from the embryonic (amniotic?) sack. This would all need to be done under temperature-controlled sterile conditions of course, and the process would also require just the right mix of nutrients, enzymes and hormones.

Can anyone point to someone who has demonstrated success with embryo rescue of cannabis seeds that are 40+ years old?

Im on the same boat that's why I was researching tissue culture if you find out please let me know I shall do the same if I run into anything.

 

[gmanwho]

 

[gmanwho]

I'm sitting on a few grams of bagseed collected from the mid sixties into the early 1980s. After blowing through a few dozen trying various techniques, and only getting a couple to shoot tails, I'm currently sitting on the rest waiting for TC and embryo rescue technology to advance so I can improve my odds. They were gifted to me about 6 years ago and are the treasure of my collection, despite being currently unviable. Operative word there is currently.

It is my opinion that embryo rescue technology requires some micromechanical means of manipulating the seed to help it split open and the safely remove the embryo from the seed husk and from the embryonic (amniotic?) sack. This would all need to be done under temperature-controlled sterile conditions of course, and the process would also require just the right mix of nutrients, enzymes and hormones.

Can anyone point to someone who has demonstrated success with embryo rescue of cannabis seeds that are 40+ years old?

hey, so i took a few day tc class about 2 years back. I am just a beginner, I am no expert by any means, but i am fascinated in all respects to any sort of tc protocols. I have watched countless hours on procedures. An while my hands on has been limited i do have some idea on embryo rescue process

I am in no way an expert, but i do understand this some. maybe what i can offer can help. Or at least a direction.

The exact ratios and hormones are in my notes, which i dont have access to at the moment.

What i gathered so far is that most embryo rescue is a multi stage process. Do to how fragile the embryo is, the absolute last resort is actually removing the embryo from the shell. All efforts where made to start the seeds on there own in vitro. Only after no germination are the shells removed.

A black or dark grey embryo is the worst case. was said that the embryo is dead. But what i gather is if there is 1 viable cell, you can start that cell from embryo and perform similar procedures for callus formation.

seeds shells where lightly sanded, sterilized, then placed in a sterile nutrient solution of basal salts. If the tap roots emerge they where place into individual vials with nutrient agar. if no tap root they continue to soak. checked daily over an over, eventually after a week or so if no popped they where removed from solution and all placed on ONE agar plate with nutrient solution.

then only after a few weeks of no development the embryos are to be removed from the shell and individually placed directly on the nutrient agar. the removal is usually done under a video microscope.

check out this website. they have some sops and pretty much all of the materials you will need. my advice is to check what you need here. Then shop around a little to like ebay etc..

https://phytotechlab.com

phylotech has some articles like this too for sterilizing.

https://phytotechlab.com/mwdownloads/download/link/id/3932

heres an article on seed germination

https://tgrc.ucdavis.edu/seedgermination