No breaching, I just want to tread lightly. My computer’s motherboard died after I posted last and my laptop is connected to my microscope camera. I’ll get a new thread up soon to share info.
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Done and done! So far everything is back on track.
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I’m just going to necro this thread
I’ve been reading about androgenises using pollen microspores in media for the development of double haploid plants. For anyone who may not know, this is a process of creating true breeding variety in one move by creating a haploid plant via stamen/pollen culture and then chromosome doubling to get a direct duplication and therefore a completely homozygous variety. If you have ever had to line breed to stabilise a variety then you know it’s a long time consuming process.
This removes that process and allows for the rapid development of true breeding plants that can then be used for breeding F1 hybrids etc.
Anyone got any info on if this is happening commercially yet and if there is a protocol around it? It seems like cannabis is a reasonably recalcitrant plant when it comes to tissue culture in general, is that a fair assessment?
Thanks 
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Is this something like a clone in seed form?
Sorry for the ignorance 
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No it’s not that, I know what you talking about though, that shit has Big Ag written all over it.
No, It’s where you take a single parent; different process but same concept for males or females, you use either anther or ovary tissue and culture them into plantlets, they will then only contain genetic material from either the female if it’s Gynogenesis, or the male if it’s Adrogenesis. The resulting plants only contain genetic information from the male or female gamete so they are sterile, so typically a mutagen like colchicine is used to gene double them so you end up with a Double Haploid which is completely homozygous; you have basically duplicated all the alleles across and guaranteed a 100% true breeding variety in one single generation.
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You’re really looking into virtually uncharted territory for cannabis as far as publicly available knowledge is concerned. Genetic modification is not something that has hit a lot of publications due to Cannabis remaining at Schedule I. You can find some information if you revert back to seeking out studies on “hemp.”
You can take a look at the following paper and keep us posted on any of your laboratory results.
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Yeah, there is some research on protocols but not any that I can find that report repeatable success.
This is a good read:
This is definitely a process that is going to happen as it all becomes more commercial and industrial.
I’m thinking some of the big canna corporations must have this process as trade secrets.
In other crops there is a few different ways that haploids are made, one is by using another variety that has a mutation so that 25% of the progeny are haploids,
Another approach is to use various forms of radiation on the pollen: damaging the pollen so that it can germinate on the stigma, grow though the style and reach the embryo sac, but is unable to fertilize the egg cell/polar nuclei and stimulating the development of haploid embryos etc. Though I think this method may require embryo rescue techniques that probably don’t exist for cannabis yet.
It occurs to me that a ‘light weight’ mutagen applied on the pollen may have a similar effect, maybe a caffeine I’m thinking as it’s known to to have such effect on pollen grains and is definitely a lot less brutal than the other mutagens. Though I
would probably need to work out the best way to mix and combine them and then dry the pollen…maybe via a centrifuge and ethanol dehydration and then experiment to see how much caffeine for how long etc… ethanol might be too harsh… Not sure, just spit balling really lol.
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That’s pretty interesting, but how can you select for the traits you’re after if a stable plant is created in just one generation?
I’d like to know more about this, pretty interesting stuff. It just seams like it would bottleneck the gene pool of cannabis at a faster rate then it’s all ready happening
The process basically uses the unfertilised egg or pollen and then cultures a plant that only contains chromosomes from either the mother or father side, the resulting haploid plants are usually sickly and sterile because they only contain half the chromosomes. They then are treated with chemicals like colchicine which ‘gene doubles’ I.e basically duplicates the chromosomes across to give correct number of chromosomes; a ‘double haploid’.
This process of copy/pasting half the chromosomes to get a full set means that the plant is completely homozygous across all traits and breeds true. If you say did it to a well known cut, it would mean you are basically hard coding it so that the next generation would all be virtually identical, every seed the same. So you do the selection first i.e. find the perfect pheno and then use this process to completely lock it down, and because you haven’t inbred to get there, it means no inbreeding depression. As a process it wouldn’t bottleneck, in theory it’s just a short cut to getting stable lines you can use for hybrids.
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Oh ok, I misunderstood the other post. I wish I had a lab sometimes to try some of this stuff out lol
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Yeah, you kinda definitely need a sterile workbench with a laminar hood. Also all the sterilising stuff and the microscopes etc. and all the hormones and medium, and the incubator and a shitload of time and patience lol.
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I have time and patience lol but I wouldn’t know where to start. I’ve tried to look into tissue culture but I haven’t found anyone to follow steps to do it. I have some experience with agar but as far as recipes and the process goes I get lost pretty easy
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Stage 0: Grow healthy Plants and cut from healthy growing shoot tipsp
Stage 1: Sterilize and initiate that culture to MS media
Stage 2: Multiply the established explant on a cytokinin-rich media
Stage 3: Root the multiplied explant on auxin-rich media
Stage 4: Acclimate the shooted and rooted explant
You really only need to know how to surface sterilize plant material, make media, and how to perform the aseptic techniques.
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@SCJedi The making media is what I’m most confused about. I’ve worked with agar and mycelium but for the nutrient side of it are regular rooting/veg nutrients added to agar or is it something different used? I have the basic supplies to try this out sometime, I not looking to have a warehouse full of TC but I think it could be a massive space saver for my breeding projects.
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MS is the nutrient part. Add some sugar as the carbon source and pour your MS/Sugar mix into a vessel with dry agar and sterilize it. Once it cools a bit add a sterile PGR and pour. MS + Sugar + Agar + PGR = TC Media
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