Re: [DIYbio] Re: Want to work on creating cold resistant wheat

One issue with cereals is that they don't prove to be a good agrobacterium host even with additional stimulant (phenolic like molecules like vanillin or acetosyringone). You don't see crown gall disease on cereals for a reason. A gene gun may prove easier and there are designs online that are cheap. I made one similar to the one from the VIGGS paper here:

http://www.plantmethods.com/content/pdf/1746-4811-8-27.pdf

And it worked fairly well. I use tungsten so the buffer needs to be at a higher pH and you'll need to work cold. I get my particles from Usnano.com its the 800um tungsten. Last i checked it was $42 for 5g and 16mg gives you 7ish shots. The process is super low efficiency so the first thing I always recommend to newbies (and is seldom considered) is to start with getting tissue culture down packed. Its the hardest of the processes and will make or break your experiments. Plants From Test Tubes is a great book for beginners and a follow up is Introduction to Plant Biotechnology by Chawala...and hopefully I'll finish my book by winter (shameless plug).

You can take this article as a good basis for ICE pathway genes from Arabi:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC219064/

I work with plants and have been for a long time. If you need more info just ask. I'm an open book. Good luck!



Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC


> On Aug 9, 2015, at 8:22 AM, Mega [Andreas Stuermer] <masterstorm123@gmail.com> wrote:
>
> If it is "just" about expressing antifreeze protein in wheat, that seems fairly doable. Probably you also find a cold induced promoter out of the literature.
>
> I was told that monocots are different to regenerate from a gew transformed cells, though.
> Agrobacterium may be easier, but then your organism is regulated by FDA and USDA.
>
>
> Here is a small intro to plant trafos (transient). Might be worth a look.
> http://diyspartanbiotech.files.wordpress.com/2014/03/genetic-engineering-and-synbio-for-beginners-v1.pdf
>
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