Relatively safe-to-handle mutagens to expose seeds to?

[Warren Notes]

Does anyone know of any relatively safe-to-handle mutagens that can be legally obtained - and that are relatively safe to use - that seeds could be exposed to to encourage mutagenesis?

At one time, I thought about buying old radioactive watches off of ebay that had luminous dials. However, a university professor I asked told me the radiation from them would be far too weak to induce mutagenesis.

Any ideas?

-John W. Wilson

[Cory Tobin]

The compound typically used for inducing mutagenesis in seeds is EMS
http://en.wikipedia.org/wiki/Ethyl_methanesulfonate

I'm not going to claim it's safe. But if you have the proper safety
equipment you can use it without giving yourself cancer. You'll need
a fume hood, gloves, lab coat, and face shield because you really
don't want to get this stuff on or in you. It's not something safe to
work with in most DIY labs that I've seen.

If you have experience working with hazardous materials and have the
proper safety equipment and the ability to purchase hazardous
compounds, then go for it. Otherwise there are companies that will do
it for you.

Lehle Seeds can do it for you. They can also do radiation induced mutagenesis.

-cory

[Cathal (Phone)]

You can get cheap mutagen-sticks in packs of 20 in most corner shops. You'll need a burner and a box to contain the smoke within. Don't breathe it in, it'll give you emhysemia. :)

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[Nathan McCorkle]

x-ray tubes are probably obtainable/constructable most places (I've
definitely heard of tubes being made DIY, but I also remember it was
quite a skill to do so!)
http://www-naweb.iaea.org/nafa/pbg/public/Protocol-X-ray-mutagenesis-plants.pdf

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-Nathan

[Jeff Backstrom]

Never used them, but CSU has an irradiation service.

http://csu-cvmbs.colostate.edu/academics/erhs/pages/irradiation-services.aspx

If you want something like Co-60 gamma irradiation for seeds, you'd probably have to start emailing gamma irradiation services, and see which ones will dial down their machine from "obliterate" to the "small parcel" setting for small runs.

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[Nathan McCorkle]

On Sun, Aug 24, 2014 at 4:55 PM, Jeff Backstrom <jback...@gmail.com> wrote:
>
> Never used them, but CSU has an irradiation service.

Nice, that reminds me, Reed college also does this:

http://reactor.reed.edu/experimenter.html#Gamma

[Warren Notes]

Anyone have any thoughts on UV light as a mutagen? I've thought about that, too, but also had that idea pooh-poohed - even though I've seen research studies where they talked about exposing seeds to UV, I suppose that the commercially available bulbs are just nothing compared to the ones used in labs - is that so?

-John

[Warren Notes]

Actually, I've thought about soaking tobacco to make a solution, I see no danger in that, as it would have to be orally ingested to cause harm. Then the solution could be used as a spray to germinate the seeds and for watering. But - how toxic would be too toxic? I read a claim once that prisoners sometimes commit suicide by soaking cigarette stubs in water and then ingesting it. The article claimed that they would go out with a heart attack while experiencing a soaring high. That sounds pretty toxic. If it was a solution, could you expose the seed without killing it?

-John

[Nathan McCorkle]

On Mon, Aug 25, 2014 at 1:14 PM, Warren Notes <warre...@gmail.com> wrote:
> Anyone have any thoughts on UV light as a mutagen?

It works for e.coli, though I feel like with a seed you'd not have
such luck as the penetration depth is probably pretty low. Google
Scholar it up!

[Nathan McCorkle]

On Mon, Aug 25, 2014 at 1:24 PM, Warren Notes <warre...@gmail.com> wrote:
>I read a claim once that
> prisoners sometimes commit suicide by soaking cigarette stubs in water and
> then ingesting it. The article claimed that they would go out with a heart
> attack while experiencing a soaring high. That sounds pretty toxic. If it
> was a solution, could you expose the seed without killing it?

There are lots of other toxins synthesized in combustion during
smoking, but chewing tobacco is def carcinogenic.

[Sebastian Cocioba]

According to Ames, all mutagens are carcinogens. The UV sterilizers use UVC short wave but may not penetrate thick seed coats. Arabidopsis or orchid seeds are thin and lack opaque starches so the light may penetrate deeper. If you don't want to commit I could run some and show results.

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[Cathal Garvey]

Sebastian; would it work if you were to remove the outer coat? How often
does that just kill seeds?

If you removed the coat, possibly even removed one of the seed halves to
expose more of the embryo, you might be able to mutate the embro and
support it through germination with TLC..

Alternatively, don't work with seeds; culture a callus of
undifferentiated cells and expose those to abundant UVC, or grow on a
tobacco-infused agar medium. Callus is already pretty happy to mutate
somatically as it doesn't have the same structural mechanisms used by
plant meristems to exclude mutant cells.

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[Nathan McCorkle]

On Mon, Aug 25, 2014 at 1:53 PM, Sebastian Cocioba <scoc...@gmail.com> wrote:
> According to Ames, all mutagens are carcinogens.

Don't we know that cannot be true though? If a mutagen induces a
lethal mutation, it cannot be carcinogenic as cancer
growth-out-of-control, not death-out-of-control... I thought.

[Cathal Garvey]

The Ames test is a cudgel-like test for mutagenesis. Bacteria and
Eukarya are so different in their treatment of their own DNA, their
approaches to handling mutation and heritability, and in anatomy and
relative organ/elle partitioning from mutagens that all the Ames test
can really say is "if this stuff is in contact with DNA, it might modify
it".

Some example scenarios where the Ames test is useless:
* A substance may be teratogenic if in contact with gametes or embryos,
but will never, for metabolic, digestive or simple anatomical reasons,
come into contact with these cells or organisms.
* A substance may be mutagenic, but the types of mutation it causes are
either trivially repairable or too lethal to individual cells to cause
long-term harm (e.g. cancer)
* A substance may be mutagenic under certain conditions, such as a petri
dish, but protective under others. See also; loads of antioxidants,
which can be mutagenic in some redox states and protective in others.
* A substance may be mutagenic to bacteria because they are bacteria.
See: lots of antibiotics.

..and so on. I don't take the Ames test very seriously on its own; it's
only useful as an indicator that further study may be merited, but if
that further study turns up nothing, move along.

[Simon Quellen Field]

In the spirit of DIY, Spectech   sells radiation sources that are exempt from NRC restrictions.

Carolina Biological sells a kit of three of them .

Pasco sells a kit of 8 gamma sources  , so you have a wide range of energies to try.

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[Yuriy Fazylov]

what is the scale of mutagenesis are we talking here?

[Nathan McCorkle]

Nanoscale

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[Yuriy Fazylov]

a real question deserves a real answer

This is how you get mutation on sarcasm level ... 

[Nathan McCorkle]

On Tue, Aug 26, 2014 at 3:12 PM, Yuriy Fazylov <yuriy...@gmail.com> wrote:
> a real question deserves a real answer
>
> This is how you get mutation on sarcasm level ...
>

I was quick to respond, but we are talking about DNA here... unless
you meant something else by scale. What were you asking? number of
organisms exposed per dose, number of organisms in general, if the
organism was micro/meso/macroscale itself...

[Yuriy Fazylov]

What is your target for mutagenesis? SNP? Rearrangement in chromosomal architecture?

Chemical mutagen? Radioactive one?

You might want to familiarize yourself with the types of radiations you want to use, if you deem it safe enough.

Although this is rated as not a scientifically reputable site on WOT, some info is better than none. That is not to say that the research may not be viable at all. Some researchers get lured into something like this and have a hard time clearing their name from such sites. This article close to lists what type of radiation gives you the type of results you may want/need for your study.

http://www.hindawi.com/journals/ijpg/2011/314829/

“Compared to radiological methods, chemical mutagens tend to cause single base-pair (bp) changes, or single-nucleotide polymorphisms (SNPs) as they are more commonly referred to, rather than deletions and translocations.”

“Among the radiation-based methods, γ-ray and fast neutron bombardment now supersedes X-ray in most applications. Of these, γ-ray bombardment is less destructive causing point mutations and small deletions whereas fast neutron bombardment causes translocations, chromosome losses, and large deletions. ”

[bant...@gmail.com]

Hi John, I sugest you get the book, "Mutation Breeding" by A.M. van Harten: ISBN 978-0-521-03682.It provides most of the answers you seek. Good breeding, banta

On Saturday, August 23, 2014 6:56:48 PM UTC-4, Warren Notes wrote: