First off thanks for featuring my question on the last episode about the 12/12 from seed. To touch base on the tissue culture questions. I have a clean room area where cultures will be made and maintained, and yes I can store and create thousands of plants with a few callus masses from the tissue cultures. I’m looking to bank the tissue cultures to have hundreds of strains available without any seed(or make artificial seed with tissue cultures in the fake seed), at a moments notice. The good part is you test the plant once and it should always be the same CBD/THC levels, each time just as a clone, but without the cells being mature and the plants life cycle starts over every time you culture it.
I will be making some polyploid plants, by subjecting seeds to Colchicine derived from Autumn Crocus (a common flower which is poisonous) by crushing their seeds or extracting from the plants. This will be a big project but I think the time is worth the reward.
This has been used in the industry to create large flowers and used in breeding forever. Colchicine is used for gout treatment and over ingesting it can actually kill you, so you don’t want to be smoking the 1st generation of plants, just the ones you grow from the seed you create. It mutates the plants to double up the chromosomes and then it becomes a polyploid, not a diploid. I have read that tetraploids 4n grow slowly. So my idea is create a tetraploid, and then pollinate with a standard diploid 2n, which will make it a 3n (pretty much the equivalent of a mule, cannot create offspring sort of ) (for reference: A horse has 64 chromosomes, and a donkey has 62. The mule ends up with 63. Mules can be either male or female, but, because of the odd number of chromosomes, they can’t reproduce)
Polyploid is a natural mutation in nature in which a cells get extra sets of chromosomes(think of giant redwood trees 66 chromosomes total, I wondering if any of those extra chromosomes make them huge?). Cannabis plants are usually diploid 2 chromosome, which means that they have two complete sets of chromosomes. Those being Polyploids have a higher number of chromosomes sets, so there can be triploid (3n), tetraploid (4n ) on and on. You can keep going higher. But here comes the interesting part.
If you have a triploid (three sets; 3x), for example seedless watermelons you can have an outdoor grow that cannot be pollinated ever by regular 2n diploid pollen. or pollen from hemp naturally growing. So you could breed in a grow room and not accidentally pollinate the plants, sitting next to them. not my photos, but the general idea, this i believe is the slower growing polyploid.
A common flower that now has doubled up chromosomes. They look like aliens I know.
I believe if you were to look at some strains that are higher THC they possibly could be triploid 3n plants now. which would explain if you are having issues breeding other strains to it, and the ever increasing THC levels.
https://en.wikipedia.org/wiki/Polyploid
I will publish videos on this also. I have some research on this subject along with the tissue cultures for a number of years just just started growing this year. So I will be posting lots of info online.
Love to hear you take on this. I know I have some ambitious projects, but there are hundreds of haters that will say 12/12 from seed is crap, and I bet with some strains sure. But when I find that strain I can produce a lot and have constant flower coming out I’ll be happy.
Also I will share my creations with giveaways, with the community.
thanks, MaineGrower
This article on hightimes re-enforces my ideas, just think bigger flowers/double triple size?.. possibly higher THC/CBD.
http://hightimes.com/grow/is-mutant-weed-on-the-horizon/
also it says it doesn’t consider this GMO, because no foreign DNA is introduced in.
I think I’ve made a monumental breakthrough in polyploid cannabis breeding. I need help to keep my discovery from being stolen/patented by a corporation. It could be huge…literally. I’m talking oak sized plants that could even potentially be sterile…true sensi, non-gmo. This would have applications in phytorestoration projects, hemp lumber…truly monumental. I don’t know how to get in touch with you.
Dhganja,
I am working on this as well, and have guidance from some pros, but also feel that we should be sensitive what direction we punier this tech… Get in touch with me, would be good to connect, even just to nerd out. laredjeach@gmail.com.
pioneer this tech**. It absolutely needs to be protected from corporations.. and patents! As soon as something is patented it allows them to lock it up and prevent it from being a world saving existence… to keep the money in their pockets and us scratching our heads thinking that ‘solutions’ don;’t yet exist.. smh.
In my opinion, intentionally inducing mutations is something that we shouldn’t be messing with. I understand that this has been done with fruit ect. in the past but that still doesnt make it a good idea. Maybe no “foreign DNA” is being introduced, but you are introducing a foreign chemical with the intention of causing mutations. To me, this is arguably still genetic modification and creating GMOs.
There is nothing natural about intentionally inducing genetic mutations with chemicals, even if those chemicals are plant derived.
I also just gotta say I think playing with chemical mutagens in the pursuit of bigger or more potent buds is unnecessary. There is still so much to be gained through traditional breeding methods! Just think about how much popular cannabis varieties have changed in just the last few decades. Now imagine what the next few decades will hold now that the barriers of prohibition are falling and breeders are able to work more openly…
I dont want to discourage you or anyone else from experimenting or learning, but I think its important to recognize that we are talking about inducing unnatural and unpredictable genetic alterations to a plant that many of us consider sacred and use for medicine. While I get that some people are excited by this prospect, many of us find it completely horrifying.
I’m working with CBD varieties and if I can mutate and enhance the usage of the CBD I’m going for it. My first though lot was it sounded rather Monsanto. Because it’s the direction they would take
http://m.amjbot.org/content/101/10/1711.full
This was a decent read. As well as this https://blogs.scientificamerican.com/artful-amoeba/for-plants-polyploidy-is-not-a-four-letter-word/
Tons of other articles showing polyploidy naturally occurring in nature also with evolution of plants.
Thanks for the input. I’m still experimenting either way, but I love be to hear comments about ideas and thoughts of it.
I wonder if you could breed two of the polyploid plants together and get a seed with the genetics. It would be interesting to test and long as you are mutating anyways you might as well “male out” one of the ladies with coloidal silver and get some feminized poly seeds. I like experimenting too, even if just to see the outcome, but I’m with Soup in wanting the most pure natural stuff as medicine/rec.
Interesting articles and I fully admit it is an interesting topic.
Just to be clear, its not naturally occurring polyploids that concern me, its the use of chemicals to unnaturally induce polyploids that I worry about.
I’m not convinced that polyploidy is the only effect or mutation that can be caused when using these sorts of chemicals and I don’t think their effects have been well studied. These chemicals interfere with the way cells normally divide and multiply, which could have all kinds of crazy implications. There may be other mutations going on or consequences that aren’t immediately clear, perhaps even for several generations. Breeding with naturally occurring polyploids doesnt worry me nearly as much, but from what I understand, it doesnt seem like a very effective approach.
I’ve seen zero actual evidence that polyploid cannabis actually results in a more potent plant or has any other desirable traits besides goofy looking flowers. I’ve read about the Buddha Seeds experiments with polyploids, but larger seeds, leaves, or cells, are not necessarily indicative of improved plant health or bud quality. Its also not clear to me if this was an isolated result or if they if these results are consistent and repeatable. Since they are using colchicine, it could even be the chemical causing these effects, not the polyploidy. My understanding is that you are way more likely to introduce instability, which would make it even harder to stabliize any beneficial traits you did happen to come across.
If your goal is increasing CBD, I would think working with established high CBD strains, doing lots of test crosses, backcrosses and lots of testing would be a way better way to go about it. or maybe even working with some old school hemp varieties would be an effective place to start. Trying to find favorable CBD traits through chemically inducing mutations seems unlikely, and I suspect they would be very hard or impossible to stabilize into useful seeds. I will admit I’ve mostly only read about plant breeding and havent done too much of it myself so maybe I’m totally wrong. Just seems like there are easier or more effective paths to the same goal.
I highly recommend reading “The Cannabis Breeders Bible” by Greg Green if you haven’t already. I dont remember if it talked much about polyploidy, but there is tons of amazing info in there. Lots of best practices for breeding and how to select for and isolate traits like high CBD.
Anyway, have fun experimenting! As a fellow amateur mad scientist I wish you luck on your adventures. Just dont forget to think through what you are doing and always have good reasons for doing it. And always be careful and STAY SAFE!
I do have The Cannabis Breeders Bible. I will have to dig it out. I also have another of his books. Mostly its for the overall experimentation. I hope to just document it and see where it goes. Heck if it ends up being a waste of time I will have at least ruled it out as a viable option. There was talk of some of the bigger strains already being bred from mutated plants, so you may unknowingly be smoking/ingesting it already.
Some different seed banks have done a lot of tests with triploid and tetraploid plants. You have to have access to flow cytometry to even know if your tissue is polyploid. It the just about the only 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. You also have to test the roots, shoots, leaves etc to make sure all parts of the plant are polyploid. I know a couple years back buddha seeds in spain did a bunch of tests inducing mutations. I believe they got better results inducing the mutations on clones from known genetics that had been flowered so they could see how the extra sets of chromosomes influenced characteristics of the mature flowering plant. Look forward to hearing the results you find and wish you the best of luck.
I found the abstract from the study: Here you go:
Appl Biochem Biotechnol. 2015 Mar;175(5):2366-75. doi: 10.1007/s12010-014-1435-8. Epub 2014 Dec 10.
Effect of induced polyploidy on some biochemical parameters in Cannabis sativa L.
Bagheri M1, Mansouri H.
Author information
Abstract
This study is aimed at testing the efficiency of colchicine on inducing polyploidy in Cannabis sativa L. and investigation of effects of polyploidy induction on some primary and secondary metabolites. Shoot tips were treated with three different concentrations of colchicine (0, 0.1, 0.2 % w/v) for 24 or 48 h. The biggest proportion of the almost coplanar tetraploids (43.33 %) and mixoploids (13.33 %) was obtained from the 24-h treatment in 0.2 and 0.1 % w/v, respectively. Colchicine with 0.2 % concentration and 48 h duration was more destructive than 24 h. The ploidy levels were screened with flow cytometry. The biochemical analyses showed that reducing sugars, soluble sugars, total protein, and total flavonoids increased significantly in mixoploid plants compared with tetraploid and diploid plants. Tetraploid plants had a higher amount of total proteins, total flavonoids, and starch in comparison with control plants. The results showed that polyploidization could increase the contents of tetrahydrocannabinol in mixoploid plants only, but tetraploid plants had lower amounts of this substance in comparison with diploids. Also, we found such changes in protein concentration in electrophoresis analysis. In overall, our study suggests that tetraploidization could not be useful to produce tetrahydrocannabinol for commercial use, and in this case, mixoploids are more suitable.
From the research I did, it had said inducing as seed is the best option because the plant can revert or change back if induced as a branch/tip/section.
for this. I will start by making my own seed stock, using known seeds of a regular variety, non-feminized, (Grand Daddy Purp) and saving pollen from a male to back cross to it, of the same variety. This will let me document the leaves/growth patterns of a regular plant and take photos of them. (I have these seeds germinating now, hoping for a good male and female out of the 5 in the dirt. to make new seeds)
Once I get saved seeds from the above 1st grow, I will subject sets of seeds to different % of Colchicine, following some directions from some plant breeding sites, as done in other industries to induce.
For knowing the plant is polyploid, everything I have seen, shows the stem fat/swelled I will keep any with a swell, and change concentrations and keep trying, until enough germinate to experiment with, that live. Everything I have found said this will kill just about every seed you try it with. (That’s why I’m starting with my own seed to have hundreds to mess with, that don’t cost me an arm/leg to obtain)
once I get them tall enough and sex shows I will flower them short, and use pollen from the diploid variety to get seeds from that, back crossing the original male pollen used or pollen from a clone of that diploid male to give me a triploid, which should be faster growing then the tetraploid and not able to be pollinated from a diploid anymore, which I can test by subjecting it to diploid pollen.
after that point I will purely just plant those seeds and look for the signs of triploid, sets of leaves/general look.
At that point I will use tissue dying and get a count on the physical leaf as the Buddah guys did or some samples out for testing.
I’m shooting for triploid for many reasons. I think its a superior crop with just enough ahead of diploid, but safe to plant near breeding projects or outside and not fear the accidental pollination from outdoor crops.
We have a 420 acre cannabis complex in southern Colorado. I would be very interested in your consultation regarding your work on polyploid cultivation. Please contact me michael@coloradoarea420.com
Cheers
Generally with a polyploid plants we may be able to get a desirable trait to show itself but the negative effects on the plant generally outweigh the postives. Think Rainman.
Growers Love Grow Guru
Exactly what I was thinking guru
Great thread guys. This was the first time I had really looked into this. I am grateful for all of the knowledge here.
these triploid plants are far superior to any diploid I have ever seen. Superior in every way. I’m incredible high off one right now and having no abnormalities. I don’t care if it’s gmo. in fact I prefer gmo fruit and bud. technology is a good thing. I will add that they dont look mutated like these pictures from 2016 anymore.. they just much bigger overall and sterile with better flavors and easier to grow.