Polyploidy

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foxd
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Re: Polyploidy

Post by foxd »

I probably should start documenting this with photos, it seems to be generating a lot of questions.

I did dig a division of Arundinaria gigantea yesterday, remove all the dirt and look closely at the rhizome before putting it in the chemical bath. I looked at them this moring when I removed it from the bath and put it in water. The rhizome buds had definitely swollen over night and it looked like more buds developed on the rhizome. Whether this is due to the oryzalin or not, I don't know. Since my experiment is to induce polyploidy, my control group is the plants not treated.

Though thinking about it, washing every last bit of dirt off the roots/rhizomes of the plant, examing the buds, sticking it in liquids for about a day and then closely examining the buds again is not something I normally do to the plants. We normally see the shoots emerge in the Spring, but we normally don't watch the buds develop underground. This may be normal.
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Re: Polyploidy

Post by needmore »

Usually, I believe, the buds that are 'destined' for spring shoots will already be swollen by early fall and slowly continue to push as soil temps allow during the winter. Recall the photo I posted last summer of a Parvifolia division, the buds had swollen and already had some color on the sheath, I do not believe these were going to break ground in the fall had I not dug them. I have dug in early January before and found lots of buds with color so it seems like either the warmth, water or the drugs are jump starting the buds you see rapidly changing.

BTW - What the heck are you doing?
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foxd
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Re: Polyploidy

Post by foxd »

needmore wrote:BTW - What the heck are you doing?
http://en.wikipedia.org/wiki/Polyploidy

I am attempting to induce polyploidy in various species of bamboo. From what I've read the various Pseudosasa are thought to be sterile crosses of other species. By doubling the number of chromosomes it may be possible to make them capable of producing viable seed. This sort of thing is done with other types of plants, like hosta, so it should be possible to do it with bamboo.

So far I have treated Pseudosasa japonica and Arundinaria gigantea. Since I have plenty of the solution, but hate to waste it, I am considering treating some more species to see what, if anything, happens. I guess I am sort of trailblazing on this. :?
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Re: Polyploidy

Post by needmore »

Well you can come out and dig some Nuda, Praecox, Henon, Heteroclada perhaps more to 'spearmint on. I am planning on a dig this Saturday anyway, I've got Nuda to cull.
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Re: Polyploidy

Post by Jeff: Igor's Apprentice »

A couple abstracts connecting stoma length with polyploidy:
After colchicine treatments of lettuce cultivars, tetraploid plants have been obtained. Crosses between 4x and 2x plants resulted in 3x plants. Between these 3 ploidy levels differences occurred in many plant characteristics which can be used as selection criteria. The number of plastids per stoma is suitable for distinction between the 3 ploidy levels. Stomatal length, size of flower, pollen grain length, seed length and seed weight can be used for distinguishing 4x and 2x plants. The percentage of stained pollen grains and the seed production per plant can be used for distinguishing 3x and 2x plants.
Abstract The suitability of stomata length as a criterion in the distinction between diploid and tetraploid rye-grass plants was tested.
From the data it appears that diploid and tetraploid plants can be separated with a large degree of certainty if the selection is based on the stomata length.
Not sure why this procedure was necessary, but it seems easy enough....
http://www.biologyjunction.com/leaf_stomata_lab.htm

I'm pretty sure you can also see plastids (chloroplasts) without staining. If my memory of biology class serves me. Not sure how that would work with the above procedure though.
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Re: Polyploidy

Post by ghmerrill »

its been a "couple" of years since bio classes.... I probalby could not remember which end of the microscope to look in!
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Re: Polyploidy

Post by Jeff: Igor's Apprentice »

I wonder what the stomata of bamboo look like, exactly.

Here's what they look like in corn:
http://www.vcbio.science.ru.nl/en/image ... ow/PL0243/

http://www.vcbio.science.ru.nl/en/image ... ls/PL0242/


From the Grass Genera of the World key at http://delta-intkey.com/grass/www/p_sasa.htm, I learned that Phyllostachys and Pseudosasa have obscured stomata. Arundinaria stomata is just described as "common". So, that is probably the genus I would start with if I were going to investigate it.
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Re: Polyploidy

Post by Jeff: Igor's Apprentice »

I missed this link earlier, but it shows the underside of a Pseudosasa japonica leaf:

http://delta-intkey.com/grass/images/pssasa51.gif

Maybe you can find the stomata which are described as "Stomata common; 27–28.5–30 microns long."
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Re: Polyploidy

Post by Paul Ont »

This is really interesting. I assume there are no results yet? I've used colchicine in the past and we have it in the lab... hmmm...

If this method fails to produce a stable polyploid (it's a shot in the dark anyway, especially with such a large population of cells). I wonder if better results could be obtained with a smaller population of cells, say a TC sample of a tiny piece of rhizome (Or whatever you use for micro-prop in bamboo)? Still a very interesting and worthwhile experiment!

You've turned me on to some interesting ideas. Protoplast fusion might be an interesting way to hybridize bamboo with their long term and inconsistent blooming!

Keep us updated on your experiment!
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Re: Polyploidy

Post by Jeff: Igor's Apprentice »

Paul Ont wrote:Protoplast fusion might be an interesting way to hybridize bamboo with their long term and inconsistent blooming!
Anyone who does protoplast fusion experiments on bamboo should be canonized in my book- even before death that is. Especially if they successfully fuse some large clumpers with some cold hardy bamboo and get some large heat tolerant cold-hardy clumpers. Hey, but I'm not picky- any type of protoplast fusion would be welcome and help set the stage.

That being said,
tissue culturing boo is also worthy and it sounds logical that there is a better chance of getting polyploidy from a small population of cells if one has the knowledge/skills/tools to do it.

I hope you will sometime bend your expertise to the cause of bamboo and keep us here at the forum posted if you do.

In the meantime, Welcome!
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Re: Polyploidy

Post by Alan_L »

Jeff: Igor's Apprentice wrote:Anyone who does protoplast fusion experiments on bamboo should be canonized in my book...
I read that as "carbonized", not "canonized" and couldn't understand why you were so against it, but then later in the same message said that you wanted it to happen. :lol:
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Re: Polyploidy

Post by Jeff: Igor's Apprentice »

Alan_L wrote:
Jeff: Igor's Apprentice wrote:Anyone who does protoplast fusion experiments on bamboo should be canonized in my book...
I read that as "carbonized", not "canonized" and couldn't understand why you were so against it, but then later in the same message said that you wanted it to happen. :lol:
You must run in other circles where I am known as "The Great Carbonizer". :violent3:

or not... :lol:
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Re: Polyploidy

Post by Paul Ont »

Jeff: Igor's Apprentice wrote:
Paul Ont wrote:Protoplast fusion might be an interesting way to hybridize bamboo with their long term and inconsistent blooming!
Anyone who does protoplast fusion experiments on bamboo should be canonized in my book- even before death that is. Especially if they successfully fuse some large clumpers with some cold hardy bamboo and get some large heat tolerant cold-hardy clumpers. Hey, but I'm not picky- any type of protoplast fusion would be welcome and help set the stage.

That being said,
tissue culturing boo is also worthy and it sounds logical that there is a better chance of getting polyploidy from a small population of cells if one has the knowledge/skills/tools to do it.

I hope you will sometime bend your expertise to the cause of bamboo and keep us here at the forum posted if you do.

In the meantime, Welcome!
Well, I would love to do this! It will, however, have to wait the 3.5+ years I have remaining in my PhD project!
I've not worked in monocot TC, but have, as I said, developed (tweaked might be more appropriate) a TC protocol from scratch fo the plants that work with. I have read quite a bit on protoplast fusion and on reading decided that if people were going to be serious about hyrbidizing bamboo this is the way to go. It simply can't feasibly be done the other way (very often, if ever). I could see something like dehydrating pollen and storing at -80C possibly working for hybridization purposes, but then you'd still have to contend with pollen viability, compatibility issue, self pollination, etc. etc. Most of these factors could be eliminated if cells of 2 spp. were simply fused!

Ah well. I have time to think about this now... Could be a valuable idea, bringing the bamboo industry further north...
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Re: Polyploidy

Post by David »

Hello All-
Is this the same colchicine used for the treatment of gout?
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Re: Polyploidy

Post by foxd »

Years ago I found a website where someone had worked out how to do recombinate DNA experiments at home. It had a lot of interesting information on how to build/obtain everything involved. But it disappeared from the web. :evil:

So far I'm starting out small with my polyploidy experiments and will probably have a lot of failures to start with as I figure out the procedures that work.

I have added an aquarium heater to the bath to keep the temperature at 80°F.

Being treated at the moment are S.m. 'Albostriata', Pseudosasa viridula and Phyllostachys aureosulcata.

The P.v. 'Chrysophyllus' treated earlier is now putting up a number of shoots.
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