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New study about polyploid

dabking

Member
This is very interesting, so what is the final goal? Bigger yields? THC/CBD? Stability in auto flower? Creating a sterile cannabis line with good THC levels to keep a genetic from being bread with? Or for just general improvement in genetics?
 

BuddhaSeeds

Member
Goal is knowledge and myth busting, but if we achive bigger yields, cannabinoid content, or something interesting that would be wonderful :)
 

Mustafunk

Brand new oldschool
Veteran
Interesting research... I'm curious to see the results. Did you used colchicine to induce the mutations?

This reminded me to one of those internet legends about strains, and how UBC Chemo was supposed to be a tetraploid created by doctore David Suzuki for the Canadian government lol.

the initial commercial tetraploid was developed in B.C. by the Canadian Government and UBC for Cancer patients. I would estimate that the THC content of polyploids to be well over 30% and maybe up to 40% if grown correctly. Fully grown mother tetraploid plants have been known to sell for $10,000. Clones have been known to sell for $1,000 in the past compared to the standard $5-7 range for diploid clones. Tetraploids are extremely difficult to clone (10-20%) unless special care is taken. The end product is far superior to anything commercially available and even compared to excellent connosseur diploid product due to the doubling of THC.

The resulting F1 hybrid seeds were then grown out and used for cancer patients at UBC. Somebody stole one of the clones that the Canadian Government and David Suzuki had created. It became famous and was then re-created and sold out east as Diesel and as OG Kush on the west coast and as ChemDawg in Colorado. These were all colchicine treated plants based upon their work and the popularity of UBC Chemo.

If your plant does not have four sets of leaves at the nodes then it is NOT a true polyploid plant and it not the real deal. UBC Chemo is a polypolid and had four leaf internodes. These tetraploid cuttings have different names but are believed to all be the famous polyploids named OG Kush, Pink Kush, Chemdawg, Diesel, Master Kush etc. These are really heavy feeders due to the double size buds and double the number of leaves and buds and THC glands.
 

JointOperation

Active member
so .. if these are sterile.. does that mean. you guys are workign on lines.. that will not take pollen.. meaning you could make fem seeds.. and set it and forget it. and get no seed?

perfect for guerilla grows ..?

and i read something about the potency increase up to 40% thc? is this really possible . has it been seen or tested? or ?

and when doing tests that read that high.. how many times can you reproduce those results is the question. because ive had seen buds test hit 30%.. and then they retest at 25%.. so it can vary alot from piece to piece.
 

MJPassion

Observer
ICMag Donor
Veteran
Interesting research... I'm curious to see the results. Did you used colchicine to induce the mutations?

This reminded me to one of those internet legends about strains, and how UBC Chemo was supposed to be a tetraploid created by doctore David Suzuki for the Canadian government lol.

Mustafunk,
Do you happen to have a link for that quote?

I find it very interesting and would like to add it to another discussion, or maybe you could.

I'm curious about what the OP thinks of the quote as well.
Seems to me that it's filled w a bunch of misleading garbage (myths).
 

MJPassion

Observer
ICMag Donor
Veteran
This is very interesting, so what is the final goal? Bigger yields? THC/CBD? Stability in auto flower? Creating a sterile cannabis line with good THC levels to keep a genetic from being bread with? Or for just general improvement in genetics?

#1 Goal in bold. ;)
It all about the Benjamins* yo!
*or your favorite fiat currency

What other goal can there be in this capitalist world?

The science is interesting.
I'm going to watch & see how this plays out.
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
This reminded me to one of those internet legends about strains, and how UBC Chemo was supposed to be a tetraploid created by doctore David Suzuki for the Canadian government lol.

Not true, just a legend. I asked David myself.
-SamS
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
First of all, triploids are not sterile, that is for sure. I have made many, all set seed.

Second did you confirm with a plant cytology lab that all the plant cells, roots, shoots and flowers were 100% tetraploid or triploid or whatever? You need to check all plant parts.
We found mixes of ploidy, Aneuploids, in many Tetraploids made from Diploids or with Tetraploids bred to Diploids to make Triploids. You need to have ploidy confirmed by a lab that tests all parts of the plant, roots stems, leaves, flowers.
I did find Triploids that seemed a little better as well as Triploids that seemed a little or a lot worse. To me it is not a magical path to quality, but it was fun to try.
The easy way to make Triploids is to make a Tetraploid of a great Diploid male and use the Tetraploid version to hit great female Diploid clones, all the resulting seed will be Triploid.
I have never tried to take female Tetraploid clones and STS'ed them to make pollen that could be used to make all female Triploids. Try it and let me know what happens, I don't want to do this but am curious what would happen.

-SamS
 

Mustafunk

Brand new oldschool
Veteran
Thanks for the info and contributions Sam, I imagined it after reading all the bunch of misleading claims and inaccurate bullshit. Let's call it "cannabis folk and myths". :)

Looking forward to see more info from Buddha Seeds guys. Researching and have a better understanding of the plant it's always interesting.


PS: http://billybudd.zappersoftware.com/polyploidy.html

Peace all.
 

BuddhaSeeds

Member
yes, we used colchicine; it is quite effective. is a good way but not the only one.

As far as we have seen the triploids are making seeds, but we are not sure yet of the viability of those seeds.

Of course we checked with a lab; cytometry is the only way to be sure of the polyploidy.
we find lot of mixoploids, few true polyploid but some are stable enough, they are still polyploid after several months; and their offspring are polyploid too
 

BuddhaSeeds

Member
To me it is not a magical path to quality, but it was fun to try.


I completely agree; after seeing some results, they are not magic, but we expect polyploidy can be another breeding tool, as some plant characteristics are for sure affected.

Female clones with STS can make pollen, same as diploid plants.
 

BuddhaSeeds

Member
Little graphic about how polyploids are made


eKu7b3T.jpg
 
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BuddhaSeeds

Member
Here you are a little explanation of the procedures we followed to make tetraploids:

Method 1: One of the most used in the bibliography forms is direct immersion of the seeds. Concentrations and exposure times must be very controlled, at low concentrations is not achieved the desired effect and with high concentrations plants do not germinate or die during the first weeks. The effects of the treatment cause a stop on the growth of plants, thick stems and roots very damaged (root to support treatment is a crucial factor). Past few weeks the plants recover and grow back.

r2AAlZ5.jpg


Appearance of the treated one week after treatment plants.

Method 2: Another form is to germinate the seeds and remove its cover when they begin the substratum. When the cotyledons are opened and before the first pair of leaves can be seen they are treated with a solution. Agar-agar is added to prevent evaporation of the drop; in this way, we allow that the action of the product last longer.

The effects are in this case similar in the shoot but root lesions are avoided. The plants stop growing for two weeks and spent this time the apical meristem of the stem back to form leaves (photos method 2).

YvXQhtJ.jpg

Applications : Massive treatments in the desired population.

vU3GkPz.jpg

Product Application: Detail of the treated plants.

1WXxGDP.jpg

Phenotype of treated plants , you can see how growth stops relative to the control plants.

hdvE3GD.jpg
 

BuddhaSeeds

Member
As for the methods of analysis, it was necessary to be sure that the individuals were completely tetraploid. We have put a photo with a microscopic preparation of root cells, it can be seen that the two sets of chromosome have doubled correctly but there is to look at all parts of the plant. To accomplish this we have sent samples to a laboratory for analysis by flow cytometry.

jK1o643.jpg


Flow cytometry is an analysis that quantifies the total genetic content of many cells in a particular tissue and this is the most precise method to ensure whether or not an individual is tetraploid.
 

Mate Dave

Propagator
ICMag Donor
Veteran
Feulgen stain crushed root tip and take a photo so we can see the chromosomes rejoining on the individual plants. We need to see the parents too.

I have read about people freezing plants with liquid nitrogen and then grinding them down, then throwing the powdered plant onto a flowering female and the resulting progeny had the ground plants DNA within it. Amazing! You don't need pollen..
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
We used a chemical to double the chromosomes and did confirm with flow cytometry our Tetraploids and Triploids. We did not use seeds or small seedlings, we transformed the top meristem of a clone and had better success then with seeds or small seedlings, the other advantage is we knew the potential of the known female or male Diploid clone transformed to Tetraploid better then a Tetraploid made from seeds or a seedling that has not yet been flowered.
We used elite selected male and female clones to make into Tetraploids & Triploids.


-SamS
 

Mate Dave

Propagator
ICMag Donor
Veteran
We have already done, tetraploids can be crossed with diploids, making triploids, triploids should be sterile, due to the impossibility of dividing 3 pairs of chromosomes to develop gametes.

Have you tried selfing the plants to get the stage 1 mitosis, this should work. Things that are linked will split together and discard the incompatible alleles. You will revert back possibly who know's try it and see.
 

BuddhaSeeds

Member
We also worked with clones, but as far as i understand the problem is the top meristem has many more dividing cells than the seedlings, or the soaked seeds, so the possibility of having mixoploids is bigger; also the polyploidy was not so stable, and reverted many times.

The good thing working with clones has to be the possibility of comparing the 4n plant with its 2n twin.

We decided to use the more stable genetics we could get, so we are working with s1 and cubings of the best mothers we have.

The final goal is to do it with in vitro tissue; but after achieving some results and see if there are interesting results in polyploids.

Anyway working with as stable seeds as possible and making lots of them allows to make good selection too and saves us some space, as we can easily grow 1000 seeds, but making 1000 clones needs much more space.

But for sure, now we know Sam already did it with clones and had success we will give it another try :)
 

Mate Dave

Propagator
ICMag Donor
Veteran
How did you apply the Colchicine? As far as I am aware once the microfibril is mutated it don't revert or at least it shouldn't if the mutanagen has been applied correctly. My tutor showed me his technique which he invented over 40 years ago, he has some propagules that he's patented from the process, they have never reverted.


Is the seed soaking you describe priming seeds with the mutagen?
 
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