Re: [DIYbio] Callus with Plant Tissue Culture

 I would grow the callus on plain media.  MS basal salts, phytoagar, 3% sucrose, no hormones. 

So a callus will be able to self-differentiate into normal shoots without hormonal induction?

Not very efficiently.  On plain, hormone-free media you just grow the callus to a much larger size.  That way you can split it up, expand it to cover more plates, etc.  If you want shoots and roots you'll need hormones.  You might get some shoots without hormones but it will work much better with the right hormone mix.

 
That *might* work but it will also mess with the seedling.  So I would grow the seedling and callus separately if you want the seedling to continue growling normally.

Out of curiosity, how will it mess with the seedling? I have some Drosera aliciae for example, seedlings that are much too small to separate from what appears to be developing callus cells.  

Depends on which hormones you're using and how Drosera responds to it.  Cytokinins, auxins, GAs, etc are all critical for proper developmental patterning and environmental responses in plants.  By introducing hormones to the media you short circuit signaling pathways that are critical to the plant.

In my experience with creating callus from Arabidopsis seedlings, the seedling never lives.  But if the health of the seedling is not an issue and you can't separate the callus, just put it on a callus inducing media, get some callus, (optional) transfer to plain media to bulk it up, then go to shoot and then root inducing media.

To answer your original question which is "will the callus growing on a seedling create more shoots with the addition of hormones?"  I would imagine if the callus is big enough you could get some shoots to initiate with the right combo of hormones.  But different species of plants have all sorts of weird reproduction and growth habits. So, maybe, maybe not?  I have some plants on my balcony, Kalanchoe daigremontiana, that grow somatic embryos on their leaf margins.  Imagine a human growing clone babies all over his or her arms and legs :)  Some plants behave weirdly. I don't have any practical experience with carnivorous plants to have much intuition about this, though.

-Cory

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