Re: [DIYbio] Plant Engineering

I remember in hybridoma techniques we fused mouse cells with
polyethylene glycol (PEG) of a few thousand molecular weight (MW), it
looks like it's the same for protoplasts:

a few paragraphs on protoplasts here:
http://www.rocw.raifoundation.org/biotechnology/MScBioinformatics/animalbiotechnology/lecture-notes/lecture-08.pdf

looks like a whole tutorial:
http://www.eplantscience.com/index_files/biotechnology/Plant%20biotechnology/In%20Vitro%20Culture%20Techniques/biotech_protoplast_fusion_hybridization.php

and finally a whole book with protocols references (published 1985):
http://pdf.usaid.gov/pdf_docs/PNABD686.pdf

got all these by googling:
fusion peg "plant biotechnology" protoplast filetype:pdf

also these:
http://www.btxonline.com/pages/FAQ.html#s

and another book, "Plant Cell Electroporation And Electrofusion Protocols":
http://depositfiles.com/files/37hsb7x03

On Thu, Jun 28, 2012 at 12:10 AM, Ethan <argentumxy@gmail.com> wrote:
> Greetings everyone!
>
> I have not been on here is a while as I have been excessively busy with
> school and academic research. I am curious if anyone has any experience with
> engineering plants. My family has grown our own vegetables for as long as I
> can remember, and I would like to see if I can bring anything new to the
> garden. I realize that genetic engineering through Agrobacterium
> tumefaciens or gene transfection if a bit out of range for a DIYer, but
> there are some other techniques I am curious about. I know that there are
> several compounds that can be used to induce polyploidy. Some are hard to
> come by, but it seems that Oryzalin, a microtubule disruptor is sometime
> used as an herbicide. In fact, according to one study I came across, it may
> be more effective than colchicine, which appears to be the standard in the
> industry. Paper
> here: http://hortsci.ashspublications.org/content/43/7/2248.full
> I know polyploid plants tend to have exaggerated fruits, but do you know of
> any other features I may turn up with if I attempt this (besides sterility
> with odd-number ploidies).
>
> Another technique I am curious about is protoplast fusion. I was able to
> find some nifty instructions for home micropropagation by digging through
> some old
> posts: http://www.kitchenculturekit.com/StiffAffordablePTCforhobbyists.htm
> That covers the culture part, but I am a bit lost on the whole fusion part.
> I did some protoplast experiments a while ago in a school lab, but I no
> longer have the protocol anywhere to be found. The basics seem pretty
> simple. Isolate the protoplast from cell walls by mechanical or enzymatic
> means, then you can use electricity or some chemical reagents to induce
> fusion. Would a crude lysate from some E. coli function as a decent
> substitute for cellulase? And what chemical means are there to fuse the
> protoplasts? Thanks for any help or direction you can provide!
>
> -Ethan
>
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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