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The Corroboree

Growing micropropagated plants larger after rooting, and some general observations

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Thanks again!

I will post more/ask more soon, but wanted to reply to a question asked earler.

Here are some bad photos of the rooting of micropropagated M. Speciosa plants obtained by adding honey and coconut water to the medium.

Honey was from the grocery store, said to be pure raw honey but who knows?

Sry for delay, I had to do a bit of background checking to confirm my memory was correct

How much honey/ how much coconut water did you use per litre of media?

Raw honey would be interesting, tho I suppose the autoclaving would knock the enzyme activity out as much as it would processed honey.

All the reading I have done has suggested coconut water has a stronger cytokinin activity than auxin.

Now the important factor with hormone interactions is not just how much of what hormone in which species, but also the ratio between the hormones you add in combination. So maybe the interplay is important with the coconut water. Or maybe it's some other aspect of coconut water which is beneficial with rooting. About to give it a go in some recalcitrant species, may have more info in a month or so

Those biological indeterminates can be lovely but so elusive to pin down, but shonman your experience shows that addition may have some benefits for rooting as well

Inspired me, so I'll try it anyhow :)

Don't worry about the quality of your pics, they look pretty bloody good to me. Getting publication quality TC pics is super hard, carn, we all know that. Yours are fine!

Edited by Darklight

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One question I have which I did not put very well earlier is this:

Say I have induced shoot proliferation at the axial buds at an increased level with a hormone.

Now, will I get a finished plant quicker, if I let these shoots grow longer attached to the main stem, with the rest of the shoots and the top

and then separate them later, allow them to recover, grow and root.....

Or, would I get them further along/faster by separating these newly proliferated shoots from the main plant when they have a few nodes,

so that they grow into a separate plant on their own, starting as soon as possible, like when they have three nodes perhaps??

I understand there is a recovery time after they are divided, but they would be divided eventually anyway...

do I produce plants faster, by leaving the new branching out plants on the main stem,

or, dividing them sooner, allowing them to recover and grow separately?

Took me a while to work out what you meant, thanks for clarification

This is a *suck it and see* question, individual to each species, media, growing conditions etc. These are the questions rarely specifically answered in scientific publications and need to be tailored by each lab for their own requirements.

You could do a side-by-side experiment to see what works for you, but it would only be an important consideration if you had, say, a commercial contract for replication with a definitive and punitive time line that was, say, six- eight months away ( to give you time to run the experiment and change replication protocols )

One other thing I have been wondering about...

Is there some way to contaminate plants send out to market, in agar

so they will be fine if 'deflasked' but cannot be used to produce functionally sterile cultures from/ would be contaminated 'in vitro'?

Hee hee :) There was a lot of speculation on this in the late 90s- ways to maintain IP, mask TC media so plants were still saleable but the media wasn't suitable for analysis ( if it ever was )- addition of food colouring etc. Not sure where it got to, didn't keep up with it

Not sure about your functional contam to maintain IP and commercial advantage. Short answer- I wouldn't try it. If there is a fart in an elevator's chance that whatever contam was in there could harm a worker ( spores, people with lower immune systems ) or an ecosystem ( including agriculture ) or a business I wouldn't touch it with a pole with a condom on the end. Nice theory tho

If you have developed a distinct variety you want to maintain you could patent it and protect that patent by having the plant fully sequenced ( sequencing costs about $1k last I checked but prices dropping all the time ). Patenting is a few $k too, and last I checked a patent needed to be applied for in every country the plant is sold.Then you have to enforce the patent by maintaining intel on whatever it is coming out on the market, a full time job for someone

I am in favour of patents on plant varieties which are very distinct and have been developed by the patent owner- particularly ornamental species. Some scummy companies were trying it on for older strains of food plants as a kind of astroturfing, which was a bastard act of the lowest kind. Patenting is an ethical issue so YMMV

Thanks mate for keeping us up to date on your progress, you have raised many many good scientific and ethical questions which are relevant for both beginners and experienced people alike, and you're also helping my professional development as well :D

I did check the TC email list too, on your advice, and ended up signing on again, the volume there has definitely shrunk since I was a member last time, it's down from about 40 messages a day to 4-5 a week. Most manageable, and the search function on the archives is still pretty OK

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Any information on autoclaving coconut water vs filtering it/cheap ways to filter sterilise?

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Any information on autoclaving coconut water vs filtering it/cheap ways to filter sterilise?

Coconut water which is sold for TC is heated to denature the proteins then filtered, there are teks for it. And for standard TC use people will do this with coconuts they source raw ( there was a coconut water shortage back in the late 90s sometime and labs were preparing their own )

But people can add whatever they want to TC media and prepare it however they want

I can't imagine either raw, unprocessed coconut water or commercial TC coconut water making it through any filter which would sterilise it sufficiently to add to a TC media post autoclave ( removing the need to autoclave the CW and preserving some of it's raw qualities ) but perhaps if you added it to media and filtered the whole lot it might make it through the standard filters we use these days. Is the difference between filtering the 100-200ml of coconut water and adding it post-autoclave and filter sterilising the whole litre or more of media

A solution of sterile gelling agent would need to be added post-filter to solidify as it too would not make it through a filter. You wouldn't need it if you were doing liquid culture, but you'd want really good tek because liquid culture is so prone to contamination and bugs love coconut water

Interesting thought, but too impractical for use at a home level. I couldn't even do it here cheaply. I can sterilise 10ml of a reasonable solution for about $4 if I can source a syringe filter right for the job

0.22uM is the standard filter for sterile technique at this point in time, gets out everything but viruses

All of this is subject to change the minute someone invents something new and cool ( like a super cheap microfilter system which doesn't cost thousands and clog up a lot and take expensive filters ) which subverts the current standard of autoclaving for sterilisation

Short answer: you wouldn't filter sterilise coconut water now at the concentrations generally used in TC. Coconut water sold for TC has already been filtered, but only to bump out precipitates- it's not sterile and is AFAIK always autoclaved with media

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