Author Topic: Doubling the Chromosomes of Phalaris Arundinacea.  (Read 115 times)

Vesp

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Doubling the Chromosomes of Phalaris Arundinacea.
« on: January 10, 2009, 09:18:03 PM »
See pictures with instructions in PDF at the bottom!

How to Induce Polyploidy in Plants & Seeds via Colchicine.
Introduction: Polyploid plants often have desirable traits such as increased growth rate, larger
blooms, higher alkaloid content and other qualities compared to the non-polyploid counterpart.
Inducing polyploidy is also one method to overcome the sterility of a hybrid plant species.
What you'll need:
• Solution of Colchicine
• Plants, or Seeds
• Jar (to soak the plants in)
• Water
By Way of Established Plants.
Step 1.
Soak the plant(s) upside down in a solution of colchicine. It is important to make sure you do
not soak them to long, or let the colchicine solution touch the plants roots as doing both the former and
later may kill the plant. If you did not soak the plant long enough only part of it will become
polyploidy.
Since each type of plant is different, and the assumed homemade solution of colchicine varies in
concentration as well, advice on how long to soak the plants cannot be given. Other variables will
apply such as temperature and tonicity.
Step 2.
Once the plant has soaked in the solution for the proper amount of time, take it out and rinse it
once briefly with some clean water. This will take off the impurities found in the colchicine extraction
which can cause problems later.
Step 3.
Now that the plant has had the the colchicine solutions rinsed off, place it back into its normal
growing conditions. Its growth will be stumped for several days because of the colchicine.
Step 4.
Since colchicine did not get on the roots, your plant isn’t completely polyploidy. In order to get
a completely polyploidy plant wait until it seeds or take a cutting. If you take a cutting, root and
transplant it in to suitable conditions to assure its survival.
By Way of Seed.
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Step 1.
Soak your seeds in a solution of colchicine. It is important to note that the faster the seed
absorbs the solution, the less time it will need to be soaked. Seeds that are resistant to absorbing the
solution may be nicked to encourage absorption. Soaking the seeds longer then needed will result in
death. It is good to make sure the germination rate has been significantly lowered. A lowered
germination rate will mean some of the seeds were killed by colchicine. This suggest that the other
seeds that germinated have been exposed to a high level of colchicine and are more likely polyploids.
Step 2.
Plant the seeds in the appropriate conditions. Since germination will take longer then the
average seed, and the seeds will be weaker sprouts, it is important to prevent fungal problems. I have
used pepper, cinnamon, and copper sulfate to prevent damp-off, which is caused by certain types of
fungi.
By Way of Rooted Cutting or Sprout
Note: I have personally had very little luck with this method. The only time one didn’t die with
this method was with a small rooted phalaris arundinacea, which is a very hardy plant.
Step 1.
Soak the rooted cutting or sprout in the colchicine solution. Smaller plants work better then
large plants.
Step 2. 
Rinse off the plant, put it in water, or in soil.
« Last Edit: January 22, 2009, 04:52:39 AM by Phyto »
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lqdtrance

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #1 on: January 11, 2009, 12:50:27 AM »
I am very excited to see what happens. So, how exactly do you see the chromosomes? Microscope? Can this be achieved only through cuttings or can it be trained this way and eventually seed this way as well?

Vesp

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #2 on: January 11, 2009, 07:11:39 PM »
Yes a light microscope works, It needs to be a decent one though. This isn't the best picture, but you see something like it when you are able to count the chromosomes http://www.lima.ohio-state.edu/biology/images/metaphase.jpg

I'm not to sure what you mean by trained, but once this plant has double the chromosomes, it will remain that way and all of the seeds it produces will also have double the chromosomes. You can also soak seeds in colchicine to double their chromosomes. I am going to attempt this in a while.

Phalaris Arundinacea can have a chromosome number of 14, 27, 28, 20, 30, 31, 35, or 42.
So by doubling, I could be getting some that has as much as 84 chromosomes if I were lucky. I may be mistaken, but I believe the most common is 35, so I'm likely to get 70.
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Vesp

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #3 on: January 15, 2009, 05:35:36 AM »
Update:
The grass is still alive, but it looks like about half of it died. I think this is good because that means the grass that is living got a really high dose, and so probably has its chromosomes doubled for sure.

I also let several seeds soak in a fresh solution of the extract, and also repeated it with some more grass in a moss pellet. I let a single rooted piece of grass soak in a test tube for an hour in it also, I doubt it will live but maybe.  :)


Something I want to mention is that although it is difficult to even come close to counting the chromosomes with a light microscope, you can compare and contrast how much is in the cells, and THAT allows you to determine if they doubled or not.
« Last Edit: January 16, 2009, 03:45:56 AM by Phyto »
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LYC

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #4 on: January 17, 2009, 08:12:07 AM »
Have you looked into performing somatic cell fusion? I am not an expert on it but it seems like it could be advantageous to getting a few interesting strains! I believe it is also called protoplast fusion.

Here is the Wikipedia: http://en.wikipedia.org/wiki/Protoplast_fusion

Vesp

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #5 on: January 17, 2009, 08:44:07 AM »
I haven't looked into that, thank you for bringing it to my attention.

 It seems like it would be fairly difficult to do however. I don't know how I would get the cells to grow into a plant very easily. Everything else seems like a very real possibility. I will look into it more later but I seem to be having troubles in finding very much information on it.  You are right though that it does seem to have a lot of potential for getting some interesting plants!
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cft

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #6 on: August 27, 2009, 09:45:30 AM »
So what happened to your chromosome doubled phalaris? Did it all die? Did it grow bigger than normal?

Vesp

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #7 on: August 27, 2009, 09:47:34 AM »
I got lazy and let it die.. but they did grow slow, then snapped out of it and grew at least like normal plants for a while..
But then I stopped watering it and it died haha.
I just decided I didn't care about phalaris since it contains so little and is toxic. Also kind of ran out of room, motivation, etc.
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cft

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #8 on: August 27, 2009, 09:51:59 AM »
Hmm did they show any abnormal growth or anything? Never known anyone to actually try the colchicine trick except aged hippes telling stories of when they did it back in the 60s haha.

Vesp

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Re: Doubling the Chromosomes of Phalaris Arundinacea.
« Reply #9 on: August 27, 2009, 10:04:51 AM »
I honestly can't say, I kind of wish I had stayed more with the project but IIRC it was in the winter.. so it was pretty difficult to do anyways.
I will certainly let you know if I ever go to attempt it again.  You could try it as well.
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