Sciencemadness Discussion Board - Kidney beans, Phytohaemagglutinin, and plant growth?

Kidney beans, Phytohaemagglutinin, and plant growth?

kclo4 - 19-11-2007 at 15:15

Alright well red kidney beans contain a Mitogen called Phytohaemagglutinin.

http://en.wikipedia.org/wiki/Phytohaemagglutinin

Mitogens cause cells to go into mitosis. is this the proteins function in beans, for germinating?

Also would grinding the red Kidney beans to a powder, soaking in water, filtering, and then soaking the leafs of plants in the water cause the plant cells to go into mitosis? doing so would cause the plant to grow faster correct?

What effect would a colchicine/Phytohaemagglutinin solution have on plants?

Colchicine works as a spindle poison, and makes plant cells, and other cells double their number of chromosomes because it inhibits the division of the microtubulers or something like that (not real sure as how to word that... )

So, from what i have been able to gather, the solution of Phytohaemagglutinin and Colchicine would perhaps make the Colchicine more effective and get more of the plant cells to become polyploidy, at least faster?

Anyways, if doing a simple water extraction wouldn't work to take out some of the Phytohaemagglutinin, regardless of what other molecules it brings doesn't work, how could one go about extracting it?

I read some of the DNA extraction thread, but that didn't seem to help much here, so thanks for your replies in advance cya :]

ssdd - 20-11-2007 at 18:27

Now, many hormones that cause plant cells to divide faster have limitations as to concentrations. Most of the time if you get them to high the plants pour too much energy into cell division and will die off. (I have killed a few plants this way.

) But if it is done at a decent concentration it should speed up the growth of the plant. It would be neat to try, I may have to.

As for the mix of the two, part of me wonders if they would cancel out. I wonder by what action Phytohaemagglutinin causes cell division, because the Colchicine may have just the opposite effect. It may also be interesting to try a varied mix of the two. (Not 50-50 but also 25-75 and 75-25 etc...) But again I fear this may kill the plant first.

But I think you may be on the right line in using a mix of the two as long as they don't cancel.

BTW whats your source of Colchicine? Sounds like a neat material if handled correctly.

-ssdd

BTW what are you trying in doubling the chromosome counts? Sounds neat regardless.

kclo4 - 21-11-2007 at 15:04

Well, i have been getting my colchicine from Autumn crocus

so of course, everything i do is just kinda blind, I'm hoping it will work, however my concentration is unknown. so who knows.

What i have been doing is taking the bulb like root, that can be bought at walmart, grinding it into a very fine powder with a coffee grinder. Once i have ground into a powder i take its volume worth of water and let it soak in that for a day and then i filter it.
Then i let the solution sit in a dark place for a while, so all the other white tannin stuff can precipitate out. I then sucked it off.

So, if i have 1/2cup of powder from the bulbs, i add 1/2 cup of water -- i read to do this off of some marijuana growing forum... they seem to be the only place that is interested in colchicine.

I heated it a little bit and it turned into a tea looking brown solution... kinda wondering what happened to the colchicine in that?

I heated it with an electric blanket on my bed, while the solution was poured out on a plate. This was to concentrate it. but now that i think about it i probably shouldn't, good thing i only did that to a little bit of the solution.

I poured some of the liquid on some seeds and they haven't sprouted yet. probably to powerful, or i let them sit to long... maybe it just increases the germination time? that would make sense to me.

I also poured it on some lettuce sprouts, and they seemed to morph a little bit.
And last but not least i poured it on the tip of my small flowering tobacco plant, and it seems to have killed a little bit of it, while morphing the other part.

But i'll update if they do anything els

Right now i have 10 beans in a colchicine solution, 10 beans in a Phytohaemagglutinin solution (ground beans, soaked in a slightly saline solution overnight and then filtered), and then also 10 beans in just a water solution.

if they sprout, hopefully this will give me some insight as to whats happening.

i think the 10 in the bean solution will sprout earliest. the 10 beans in the water solution next, and then only some of them will germinate from the colchicine solution.

I am going to let them sit in the solution for an hour or so, until they have absorbed a decent amount of liquid and then just set all 30 beans in a wet towel to see what happens.

I will also update you guys on this.

kclo4 - 22-11-2007 at 19:29

Today i took my bean solution, put it in a test tube, and added water to it to delute the salt solution, and after an hour the solution was completely clear! instead of the merky off white it was before.

At the bottom of the test tube is some white powder.
What could this be? do you think it is the PHA?

And if so, that means some plant hormones must be dissolved in the clear solution, right?
perhaps this solution could be used as a way to speed up the growth, or slow down the growth of plants?

Anyways, interesting the white stuff precipitated that fast... any feedback would be neat.

ssdd - 24-11-2007 at 18:37

I have no clue what it would be, but I would think that most of the hormones would remain in solution. What may have happened is you somehow removed the plant parts from a suspension state into a precipitate.

But its just a guess...

-ssdd

kclo4 - 25-11-2007 at 17:50

after a bit of research, it is protein(s) that precipitated out.

Somehow salt, or other ions help dissolve proteins or at least some proteins at a certain concentration... i have heard that ammonium sulfate is mostly, or the best to use.

Sounds useful IMO

Edit:
Does anybody have the ability to find the LD50 for mg/Kg of PHA? i have looked all over the internet, and i can't find it... I've came close it seems, but i don't know if my searching skills are the best... it seems like that sort of information on this protein would be readily available....

[Edited on 26-11-2007 by kclo4]

What i have extracted from the beans!

kclo4 - 26-11-2007 at 17:47

Edit: Sorry its so ridiculously huge, almost makes it seem not worth posting?

Well... erm this is surely not pure PHA at all, its not even pure proteins. I plan to wash it with some acetone, to get it a little prettier, when i get the chance to of course.

Please note that it is actually a lot whiter in real life, however the camera and the lighting i used made it rather yellow.
But it looks really interesting in my opinion.. i almost so a honeycomb effect, i wonder how on earth that formed?

[Edited on 26-11-2007 by kclo4]

[Edited on 27-11-2007 by kclo4]

[Edited on 27-11-2007 by kclo4]

12AX7 - 27-11-2007 at 00:04

Why on God's green Earth would you post an image that is not only over one megabyte in size, but is larger than any screen it will be viewed on, and is even completely out of focus? That's just stupid, learn to edit your images!

Tim

ssdd - 8-12-2007 at 08:05

Just wondering if you had obtained any results with this yet orr if your still working on it....

Seemed interesting I am curious if you got results.

-ssdd

kclo4 - 11-12-2007 at 21:51

I have a protein powder extracted from kidney beans, but thats all so far. I tried it again but forgot about it, causing it to rot.. so it failed.
I think i need to know some great ways of causing proteins to precipitate without denaturing them, so do you guys have any ideas? Can they be "reactivated" (in lack of better words) after an acid, heat, etc has been used to ppt the protiens? i don't think they can.. but i have heard of reversible denaturation.. just not to sure how to do it

Also, just being curious about this although I highly doubt this question is appropriate, but i am just wondering if it is possible to make a "ricin substitute" from beans and barley.
This is why i think it may be possible:

Phytohaemagglutinin causes cells to go into mitosis and messes with the permeability of cells.

"Ricin consists of two distinct protein chains (almost 30 kDa each) that are linked to each other by a disulfide bond:

* Ricin A is an N-glycoside hydrolase that targets and depurinates an adenine base at the 28s rRNA gene, resulting in an inhibition of protein biosynthesis.
* Ricin B is a lectin that binds galactosyl residues and is important in assisting ricin A's entry into a cell by binding with a cell surface component.

Many plants such as barley have the A chain but not the B chain. Since people do not get sick from eating large amounts of such products, ricin A is of extremely low toxicity as long as the B chain is not present." - wikipedia

So, this is what i was wondering, if you exposed cells (or a mouse) to a solution of Ricin A, and Phytohaemagglutinin (PHA) would it do the same thing as Ricin A and B? I think it may work like Ricin, because the PHA will screw up the cell permeability allowing the ricin to enter. Perhaps for some odd reason they may need to be connected with, say a disulfide bond?

I'm betting Ricin A is easy to extract, and I'm pretty sure PHA is also easy to extract.
If it is possible to make this "ricin" type stuff, i'll be kinda worried :S

not_important - 11-12-2007 at 21:55

Quote:

Originally posted by kclo4
...
I think i need to know some great ways of causing proteins to precipitate without denaturing them, so do you guys have any ideas? Can they be "reactivated" (in lack of better words) after an acid, heat, etc has been used to ppt the protiens? i don't think they can.. but i have heard of reversible denaturation.. just not to sure how to do it
...

http://www.eng.umd.edu/~nsw/ench485/lab6a.htm

http://www.bio.mtu.edu/campbell/bl4820/lectures/lec5/482lec5...

[Edited on 12-12-2007 by not_important]