Having had take-out this evening, and knowing a bit of your background, sleep will not come easily . . .
TerraForums Venus Flytrap, Nepenthes, Drosera and more talk
Register a free account today to become a member! Once signed in, you'll be able to participate on this site by adding your own topics and posts, as well as connect with other members through your own private inbox!
Cheap Tissue Culture
Having had take-out this evening, and knowing a bit of your background, sleep will not come easily . . .
I have agar as well
. I got a 2oz container for $5 off of Amazon. Actually I tried my local Asian market, and the man told me that the tsunami in Japan had wiped out all seaweed (and consequently agar) production 
. This cornstarch was purely for experiments, because it was $1 for a much larger package. I think that if I dye it, it may be somewhat easier to spot contamination.
just don't dye it any of the colors that are typical of bacterial or mold infections like yellow, green, blue, red, brown, black... or any color in-between all of those.
I apologize for a double post, but does anyone think I could use the cheap root hormone you can buy at any garden center in the media as a substitute for some PGRs? I think Mason said that it would inhibit shoot and callus formation, so maybe I could use it when I change flasks. I believe what I have is 0.1% indole-3-butyric acid.
Just don't set yourself up to fail. There are a few things for which there are no real substitutes; and I would be willing to send you a couple of PGRs for your use -- an auxin and a cytokinin, say, 6-BAP and NAA, which are commonly used . . .
I've been doing some research on cheap alternatives to synthetic PGRs. I could probably buy some young green coconuts from a local market. Coconut water is the liquid endosperm, rich in cytokinins, auxins, and other growth hormones. It also contains many carbohydrates that won't hurt the plants either. I found this paper that showed 10% - 20% coconut water resulted in greatly increased shoot formation in spinach. The problem with using coconut water is that there is not a specified volume to use, as it is extremely variable. Also, because I am not picking these coconuts off a tree for myself, the endosperm may have already solidified into the meat, or coconut milk. I'm going to look ridiculous walking around the market shaking coconuts...
The study: www.academia.edu/1093292/Spinach_tissue_culture_improved_with_coconut_water
Edit: Another study showing the exact compounds found in coconut water: www.mdpi.com/1420-3049/14/12/5144/pdf
The study: www.academia.edu/1093292/Spinach_tissue_culture_improved_with_coconut_water
Edit: Another study showing the exact compounds found in coconut water: www.mdpi.com/1420-3049/14/12/5144/pdf
Last edited:
The issue with coconut water, is that it is, as you said, inconsistent; and the "exact" compounds vary more than many papers, or that table even suggests. Notice the variability of γ-Aminobutyric acid, for example, between a young green coconut (at 1.90 µg/mL and a mature one (at 1730.20 µg/mL) It can have stimulatory or inhibitory effects on some cultures; it can have greater auxin activity (through hydrolysis) if the media is autoclaved; lesser so, with zeatin and other compounds if the media is not sterile filtered instead. Many researchers avoid it due to that variability . . .
Last edited:
I had no idea how I was going to measure out a volume, because there is no knowing what is actually in it. Plus, I checked three markets and there are no coconuts. Packaged coconut water has been pasteurized, so I doubt any of the cytokinins or auxins are still functioning.
What do you mean by "it can have greater auxin activity (through hydrolysis) if the media is autoclaved?" I didn't really understand that.
What do you mean by "it can have greater auxin activity (through hydrolysis) if the media is autoclaved?" I didn't really understand that.
What I meant was that the active compounds in coconut water (which is generally considered an "undefined supplement" to many researchers) -- cytokinins, auxins, and the like -- are often chemically bound with other components in the coconut water; and that the process of hydrolysis, which occurs with autoclaving can "free-up" some of the available PGRs. Paradoxically, it can also serve to inactivate others.
As an aside, the most significant component in coconut water is probably myo-inositol, which is readily available in any vitamin aisle . . .
Myo-inositol, as in the Inositol I'm using for the original recipe you gave me? When I read about Inositol, there were few specific claims, but it appeared that it helped in nutrient uptake. Thank you for the explanation, that makes more sense. The chemicals hidden away in plants are astounding.
it's beginning to sound like you are way over complicating this just to save a few bucks. literally only a few bucks... you can buy a starter kit and get started right away for 50 dollars on ebay or buy a deluxe kit for a couple hundred, and if youre serious about doing tissue culture you might as well get the deluxe kit because you'll eventually end up spending that much on supplies anyways...
It's not like your just putting baking soda and vinegar together to get get some instant result, it is a science...
edit: and what i mean by that is you can't just go into your kitchen and pull crap out of your cupboards and do it right
It's not like your just putting baking soda and vinegar together to get get some instant result, it is a science...
edit: and what i mean by that is you can't just go into your kitchen and pull crap out of your cupboards and do it right
The purpose of this experiment is not necessarily to get it right. In fact, I barely expect to succeed. What I find fascinating is the problem-solving involved in finding similar chemicals in products that are readily available to a layperson. I think that this educates me more, because it involves more research into what goes into fertilizers, multivitamins, coconut water, etc., then using a predetermined kit. If I were extremely serious about tissue culture, I would certainly by one of the fantastic kits available from the Home Tissue Culture Group. However, this project is more to educate myself on the materials that go into micropropagation, as well as to practice steril lab procedure with cheaper materials that I don't feel bad about messing up with. It's a problem-solving based learning experience.
Thank you for your opinion. I really do appreciate your feedback (especially regarding the colors above), though please do not think I am simply "pulling crap out of my cupboards."
Thank you for your opinion. I really do appreciate your feedback (especially regarding the colors above), though please do not think I am simply "pulling crap out of my cupboards."
Myo-inositol is a sugar alcohol and appears in many forms; and is associated with a number of processes, including the formation of membranes and the strengthening of cell walls . . .
