tissue culture stuff

[medicinedan]

I love cloning plants. Growing wierd plants from cuttings off strange sections.

What is that jelly, clear stuff they use in the steile jars? Can I make it? Where do I get it? Does it contain hormone stuff? Or is that applied prior to insertion. Cheers

[apothecary]

Agar.

www.funkyfungus.org

[medicinedan]

cool.

'Food grade Agar 25g' This is a powder? Is it like a gelatin with nutrients? How much gello material would I get out of 25g? enough for a few jars, maybe 10 or something?

[foolsbreath]

There are different kinds of agar depending on what you wish to colonize it with such as mea (malt extract agar)fungi & moulds, pda (potato dextrose agar)yeasts, and nutrient agar for bacteria, three of the most common type, but they are also made out of blood, milk, bile salts and a host of other more specific type agents.

[Darklight]

Originally posted by medicinedan:

What is that jelly, clear stuff they use in the steile jars?

Mostly water and sugar. Some nutrients, maybe some hormones, and a usually a substrate such as agar

Can I make it?

Yes

Where do I get it?

For 'real' tc you will need to find out what the specific requirements of your plant/ plant part are. Or take a punt and start from scratch. There is a list of ingredients as long as your arm to select from, much of which is available online.

For a basic kitchen tek which stands almost as much chance of working as anything else, UTSE to find out easy methods and links to beginners TC sites. This is my recommendation as your starting point. Google anything that comes up and then ask any questions which arise from your research here.

Does it contain hormone stuff?

Sometimes

Or is that applied prior to insertion

Very very rarely. I've heard of it and tried it with GA3 ( UTSE under gibberellic acid )on sterile seed populations with no result, which may have been the seed's fault cos it was too old

[medicinedan]

does anyone have a good link for 'home tissue culture' or something. I mainly want a recipe for a good rooting medium and one for a shooting medium. I understand these are different.

[Darklight]

medicinedan:

does anyone have a good link for 'home tissue culture' or something.

There are links here and a few teks if you do a search. Before you ask questions it's a really good idea to consult the search engine here as most things have come up before

The search engine isn't infallible, but if you at least tried it and let everyone know you didn't find anything, or have questions leading on from the answers you got, then people are happy to provide comprehensive answers

[medicinedan]

nice one darklight, that post before was relatively comprehensive and got me well on my way.

[Rev]

hey

Yes i sell agar

its fine and dandy for shrooms

but ive never tested it for plants

plant TC is a step up from fungi tho the skills are mostly the same

many fungi are right at home in the lab and the process of growing in a semisolid medium on a mix of nutrients is quite natura;

with planst you are doing something quite unatural and so have to control a lot more parameters

however its not beyond the reach on anyone committed to learning. and just liek with mushrooms you dont necessarily need a schmick setup

start HERE

http://www.kitchenculturekit.com

[Guest mandragora]

 

whatever

Edited April 16, 2007 by mandragora

[Darklight]

mandragora:

If you put a seed into tissue culture medium, it is not tissue culture, tissue culture is about chopping up plant, sorry for the lecture, got one from my supervisor as well

Your supervisor was wrong, or is at very least splitting some very fine hairs.

Aseptic germination is an entirely viable starting point for TC. Often other additives ( coconut milk, GA3 )are included in the aseptic germination media that mightn't be available from culture in other conventional substrates. At what point does the inclusion of exogenous compounds in a sterile support system with a view to changing morphology stop being tissue culture? Embryo rescue? Inclusion of GA3 for low viability seed? Inclusion of coconut milk in the media to replace possibly defunct endosperm for still viable embryos not excised but nicked for stratification? Sowing of untreated but sterile seed? That's not a question can be answered by anyone conclusively.

And for most people it is often easier to start taking propagated material from an aseptic parent plant than it is to fully disinfest say hardwood tips that have been in the field and require weeks of increasingly intensive pretreatment that could well kill the plant material regardless as to whether it effectively contains the contamination. The only cases where aseptic seed germination clearly isn't appropriate is where the genetic history of that particular plant ( for example, gene pool or chemotype ) means that seed grown parent plants may not contain all the desired characteristics of that particular parent and selection for those characteristics would have to begin again

I reckon you'll love the TC course mandragora, have fun

[Darklight]

mandragora:

You can actually buy media from some companies as well, they have embryomedia, rooting media and other things. In general it is about the hormone content or ratio of two hormones. Usually cytokinins and absicin acid, depends what you want to do with it. And some agar some sugar and sometimes some charcoal.

Good point. If you're looking at purchasing preprepared media, its great to have a catalogue as these will usually have the individual ingredients listed in the forms of uM/ml, mg/ml or broken down ionically or whatever the word is, so you can make your purchasing decision better.

Sometimes they're not called germination or rooting media, or are sold as specialised orchid media- again, look at the individual ingredients in the technical section. For beginners it's best to pick a basic media, then select variables from one or two important ones you're interested in so you don't get overwhelmed: compare and contrast.

Be aware that you'll incur delivery costs that may outweigh the purchase price of your media and co-ordinate a bulk purchase with friends, or just budget accordingly. If you only want a single $3 pack of a basic media you'll still pay a flat delivery fee of up to $25 depending where you order from

[ 12. July 2005, 01:44: Message edited by: Darklight ]

[Mecrhino]

there is a yates product I saw at woolworths called CLONEX a rooting medium, ingredients: indole-3-butyric acid.