New Grower Tissue Culture

[nben]

Howdy all!

I'm wanting to try my hand at tissue culture for autos. Not to produce hundreds of plants but as a way to store genetic clones.

Anyone that has looked into it has no doubt been intimidated by the sterile lab techniques that are involved. I have had this information for a while but am now putting two and two together and am wondering if there is a way to fuze this mushroom growing method with the tissue culture technique:

http://jontrot.free.fr/champignons/culture-eau-oxygenee-Vols1-2new.pdf

Mr. Wayne found this peroxide material when researching tissue culture of orchid seeds.

If this could be combined with the information on this site there could be a workable system without even needing a pressure cooker:

https://www.omnisterra.com/botany/cp/slides/tc/tc.htm

I would want to try for callus material from leaf tissue. There is a complete step by step HERE. Pretty amazing, toss a small chunk of leaf material on the medium and it grows into a small little blob of cells that can be divided and replicated. Still not sure how to make a cell blob grow into a plant but have found that you need to add certain plant hormones to the medium to make them do what you want them to.

Sounds like a pain to do but hopefully after one get's the technique down to a protocol it would go easy. This is coming from my desire to be able to duplicate strains. Been trying with colloidal silver but getting frustrated making seeds. Male pods seem to grow without throwing pollen... Then I thought instead of growing two separate plants of the same strain I could grow one and let it pollinate itself by just changing one branch. NOPE! Hermie issues. This seems like the long way around it but in the end seems somehow simpler... I dunno

Anyone have any thoughts or maybe experience in this field? Like I said I am more interested in long term storage of strains, like living seeds. I only grow for my own use and even then only one or max two plants at a time, I have absolutely no need for many plants at one time. I thought I could just stick the cultures in the fridge and bring them back to room temp for a day or two, or until they look like they are actively growing again but haven't been able to find anything about this. It does look like the cultures store for a very long time when they have become the little cell blobs.

Have found some information about seed starting in tissue culture but in the end it just looks like a really complicated way to sprout a seed and grow a plant. I have not been able to find any reason to want to go this route, there isn't really a way to reproduce the plant that I have seen.

On the bright side I have found some supporting information with the callus tissue. It seems that once the "explant" (the leaf chunk) grows into a cellular mass the cells become embryonic, so the age of the plant reverts back to "new", not like traditional cloning methods which would cause immediate flowering and death.

 

[sunkist]

True, you could probably get around the pressure cooker/autoclave if you use a ramped-up Tyndallization process, but if this is something you'll do often, or on a large scale, the autoclave will save you time, energy, and LOTS of money in the not-so-long run.

Going from callus to sprout is problematic, and will require at least two different media.

I'd recommend taking a look at cryopreservation using liquid nitrogen if you believe you have the skills and set up for tissue culture.

 

[nben]

Ya, starting to think this might be a little overkill. I really don't need that many plants. I think I might try to experiment with it, but only on just a "hobby" level for now.

 

[People]

Maybe your plants failed to give pollen with the colloidal silver method because you're using a low ppm. It happened to me also. Then I used a more concentrated one 100ppm and it was ok. The pollen sacks opened and there was plenty of pollen.
In order to get the desired ppm you must LOWER the voltage if you start at 26v when you get about 17ppm then your CS will start releasing bigger particules because of the increased EC of the solution. This leads to a useless solution that brown-black. If you reduce the voltage before the solution start to turn brown then you can reach much more ppm. Apliyng once a day via spray to the plant will lead to male flowering with lots of pollen

 

[fryge]

about the male balls without pollen, I had the same issue with STS on my first try this year.
what I discovered was that for the male flowers to properly ripen, the sts needs to be worn off. I suspect ethylene is involved in ripening the male flowers/pollen production(ethylene is often involved in ripening of fruits or wilting of fertilized flowers etc, I also found an article about ethylene being involved in pollenproduction in rice).

so in case of sts that means 2 treatments with 1-2 weeks apart works perfect for me. after the 2nd treatment I'll see balls forming, then about 2 weeks later the sts is worn off and the balls will be ready and will open(before, with the sterile balls because I kept applying sts, they didn't open on their own, or just a little at the tip), and drop pollen. it's still not as much as a natural male, but rubbing a finger on a reversed male flower leaves visible pollen on my finger.

I think CS wears of much faster as STS(at least from what I read about it), so you could get away with keep spraying for longer, but you would still have to stop at some point and give the balls you created time to ripen without the CS interfering.

 

[Rapha El]

Nah that's BS, just spray your sts once a week, until the plant is dripping wet, from first sign of flower until the pods open. Don't spray onto the open pods, that kills the pollen.

Here's something to get you started:

If you want to keep auto genetics around simply top the plant, put the topped part into water and treat that clone with sts, pollinate a single branch and you got enough seeds for years to come.

Make sure to use a sprayer bottle that's violet/black glass (filters out all light) otherwise your silver will fall out of solution quite quickly. You should be able to get these at any lab supply store.