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Cursed

Tissue Culture

23 posts in this topic
Just now, ratdog said:

 

 

i'm sure i said this last time tissue culture was mentioned on this site, but didn't one of the big seeds co's, dp spring to mind say they were working on seeds that were in some way connected to seed culture and would produce identical clones

 

this was 7-8 years or more ago now

Some guy posted on here even longer ago i think.  I was led to believe that was his field/profession and he dabbled with cannabis because, well, he could.

;)

 

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@ratdog

 

DP did post an article many moons ago about a development of tissue culture which used a live culture encased in a sterile medium the size and shape of a seed.

 

It was touted as a clone in the form of a seed.

 

It either didn't work/wasn't viable/too expensive or got shot down because they realized traditional seeds were way more profitable.

 

But it was definitely talked about for sure :yep:

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18 minutes ago, Dodgee said:

I mean are the cultures sat on the surface of the medium or submerged?

Ok, so there are different ways of using murashige and skoog media depending on what the intended outcome is, so I’ll just talk about how I’ve used it.

 

I sterilised tobacco seeds and then germinated them on 1/2 ms media (half ms strength and half sucrose strength). The media has phytagar in it, so like jelly. The seeds sit on top so after sterilisation they go in the dark for a a few days until they germ. They then put roots into the media. You can move them from dish to jar to jar easily as you just break up the jelly and you can pull them out without damaging the roots. So they were grown in jars until they were a few weeks old. Young stretching leaves were cut off the plant and cut into the discs you see (you remove leaf edges and midrib as the cells can’t expand).These were then washed in liquid ms media a few times. The last time was in solution with agrobacterium which will, in short  transform my gene of interest into the plant cells.

 

These then get put on top of full ms media but with added phytagar, the Leaf edges touching the media. The agrobacterium infect cells and cause calluses where it infects and inserts your gene. These leaf edges touching the media and provide all of the hormones and vitamins needed for plant growth but in high concentrations, so the explants are encouraged to regenerate. BecUse a single cell contains all the genetic info you need the discs start regenerating and organising into new whole plants, but plants that now contain your gene. So sat on top the whole time until rooting and then the roots go into the media. (You encourage rooting by removing sucrose and they start hunting for food.

 

36 minutes ago, Dodgee said:

How do they breath/respire?  

They respire as normal but just at a massively reduced rate as far as I’m aware. There’s papers looking at gas exchange etc but I’m not going to pretend I really know. But I’ve had tobacco plants in jars for weeks now and they aren’t showing signs of stress. The jars get changed regularly so I think they have sufficient gases to thrive and this gets replenished when you move them to new media. As you give them sucrose they don’t need to make food so I’d assume they aren’t respiring at a high level.

 

39 minutes ago, Dodgee said:

What's the next phase mate? Will they eventually be transplanted onto a solid medium like rockwool?


They just get potted into soil and off to a greenhouse until they flower. It’s the flowering stage my supervisor is interested in.

 

 

ask questions mate, if I can answer you know I will ;)

 

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Posted (edited)

You can buy ms media online in the right quantities with everything added together (I had to make a lot of it up myself from scratch) the problem is working aseptically and keeping everything sterile. All of this has been done in flow hoods, so that makes it pretty difficult to do at home in your kitchen. I’d imagine that was the biggest problem to making it viable for pot growers. Took me a few goes in a flow hood before I got it right and didn’t just grow lots of cool fungal statins lol

 

it also takes time and cuttings are much quicker obviously.

 

Edited by Cursed
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35 minutes ago, ratdog said:

 

 

i'm sure i said this last time tissue culture was mentioned on this site, but didn't one of the big seeds co's, dp spring to mind say they were working on seeds that were in some way connected to seed culture and would produce identical clones

 

this was 7-8 years or more ago now

 

 

This one?

 

 

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I do have a couple of Q's but I'm gonna reread the previous reply a half dozen times first till it sinks in.

 

I think the answers may already be there and it's me being dense lol

 

Cheers man :yinyang:

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32 minutes ago, Cambium said:

 

 

This one?

 

 

That thread has people posting who I haven’t thought about in years. It’s nuts going into old threads sometimes and seeing some old names.

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