IMPORTANT: HPLVD

[NezA]

Here's a microscope image, not sure if it's from a Cannabis plant or not but it illustrates the layers of plant tissue I was referring to.

 

[blackpoolbouncer]

This is getting into a fresh thread territory really but for anyone interested from my understanding is its far easier to start by using a root tip iirc

[MindSoup]

19 minutes ago, blackpoolbouncer said:

 its far easier to start by using a root tip iirc

 

I had a short stint in nursery greenhouse (boss was a cunt so didn't last long) and they did tissue culture from the root tips. Might have been an idea to stick it out, could have been a handy skill to have, but I was young and she was an awful person

[BilgePump]

@blackpoolbouncer thanks for that link dude , fascinating stuff.

 

Kinda makes me wanna crack out the old still air box I made for myco tinkering! 

 

Sterile protocol with 4 dickhead cats is just a tad tricky mind 

 

[ukbudz]

@blackpoolbouncer cheers for the link, seems pretty straight forward.

 

Think I'll grab all the equipment and give it a bash seems pretty straight forward if not a tad time consuming. I've got a rhinestone og due down in 2 weeks, I'll give her a reveg if she will then take some cuts and go from there as I know she's got it as the mother I had started showing symptoms. 

 

Might worth investing in a aero clone and going from root tips as this would limit the amount of large plants needed for a large number of atypical shoots. 

[blackpoolbouncer]

Yea mate. Certainly tempting to finish my flow hood and have a go. 

 

One thing I will say @ukbudz is obviously only use cuts from one plant at a time in the aero although I suppose infection matters not as the next process removes the infection anyway. 

[Farmer Fett]

@blackpoolbouncer Cheers for the link, I saw a guy in the states doing this sort of thing but kinda gave up looking much further in to it at that point. It looks really interesting, so I'll have a read and see if I can have a go at it.

 

 

[ukbudz]

Just priced everything up, roughly the price of 2 packs of hype seeds so well worth it to save a valuable mother. Can get everything from fleabay except the plant preservative mixture .

 

That's to do around 10-20 tissue cultures at a time. Though if your into shrooms you'll probably have most to hand I'd think. 

[Cursed]

3 hours ago, fatboy77 said:

@Cursed any help? 

I’ll see if I can answer this when I get home from work.

 

from what I can see with a quick search is that the process is similar to straight tissue culture. You need murashige and skoog media, certain plant growth hormones (often in the ms media) and a completely sterile environment, preferably a flow hood. You make up the media, sterilise it and pour it in Petri dishes, prep the plants, sterilise them in a bleach solution, place them in the dishes on the media, and place them somewhere with the right conditions. 
 

It’s a relatively straight forward process but is just a ball ache because of the need for sterility. 
 

I had about three goes at tissue culture before I managed to grow something bacteria and mold free and that was in a proper sterilised flow hood.

Edited February 15, 2022 by Cursed

[Cursed]

I’d imagine plant tissue is illegal.

[ukbudz]

@Cursed did you use plant preservative mixture as according to the article blackpoolbouncer posted as it states with out this failures would be high.

[Cursed]

Just now, ukbudz said:

@Cursed did you use plant preservative mixture as according to the article blackpoolbouncer posted as it states with out this failures would be high.

No I didn’t, my supervisor liked to add as few things as possible. He was of the opinion that with good sterilisation and aseptic  technique it wasn’t necessary. But it might be worth considering in a kitchen/less sterile environment/home made hood.

Edited February 15, 2022 by Cursed

[blackpoolbouncer]

AFAIK it's strongly recommended to be used in labs never mind kitchens when doing ptc

[Cursed]

4 minutes ago, blackpoolbouncer said:

AFAIK it's strongly recommended to be used in labs never mind kitchens when doing ptc

He’s old school and just felt if you could do it without why bother adding something. Like I said, in a lab, if you’ve properly sterilised everything and are adhering to aseptic technique it’s probably not that necessary.

 

I should add we were doing transformations so we’re using antibiotic selection which reduced chances of contamination.

Edited February 15, 2022 by Cursed

[blackpoolbouncer]

I'm presuming because of the use of plant tissue and the media being protein rich it's hard to avoid contamination with all the best will and sterile tek in the world I think. 

I'm certainly no expert just applying what little knowledge I have.

 

I know from mycology and petri/slant work with agar that no bio/fungicides are needed with sterile work so my only conclusion is it is because of the nature of the work using plant material rather than culturing fungi etc on agar