[zif]

In contrast, if someone finds (and shares) an inducer male, you could pollinate a female and then simply sprout haploid seeds! In many species these actually show spontaneous chromosome doubling, so you have doubled haploids (I like to think of them as S∞) in one step. Who knows with cannabis? It may require treatment (e.g. with colchicine) to double....

I posted over in the recent findings thread that these genetics are getting worked out in maize. It’s quite possible that researchers with access to CRISPR or the like could intentionally make inducer males right now. It may well be that the mechanisms of pollen are sufficiently conserved across plant species.

Oddly, inducer males will probably seem horrible from a traditional breeding standpoint. They’re likely to produce seeds with low or super low germination. These ‘dud’ males may be the most valuable males anyone ever runs across! Crazy....

[Tonygreen]

Anther culture to make the haploid then colchisine to restore viability and make the double hap. I've attempted to make haploids but my aseptic protocol is lacking... if I had money for a good flow hood I would buy one...

Chimera has the skills and tools to make double haps... saw one other guy with a setup he was using to make tetraploids and I tried mentioning he had the perfect set up to make double haps but he ignored me ha.

Could always try irradiated pollen as an inducer if you could get a hold of some. In the us you can find places that will irradiate seed for you. Probably have to imply you were not doing cannabis work tho ha.

[Tonygreen]

Originally Posted by HaploidDoubler

Hi all, I am an undergraduate assistant researcher in a plant genetics lab. (Full disclaimer: this does not mean I am a geneticist!) Our lab's chief research is phylogeny of brassica genus. My job just so happens to be development of doubled haploid lines of brassica, focusing on brassica in the trianglle of U (i.e nigra, carinata, oleracea, napus, rappa, juncea) these guys are important because they are at model for studying polyploidy in plants. Anyway it's done by selecting buds that are likely to contain microspores (think of them as gamete stem cells!), then they are treated with mercury chloride to clean the buds without damaging the outer layer of the bud. This is done in very aseptic conditions, over a 5 hour protocol, one mold spore or bacterium will ruin everything! after 10 minutes, the HgCl2 is flushed out, I mash up the buds, put them through a refrigerated centrfuge several times to get the microspores to pellet. then I plate them in a liquid media. Then they are "heat-shocked" at 30 C for 24 hrs to arrest meiosis (inhibiting spindle formation I think, don't quote me!) and facilitate haploid doubling, than incubated at 21 C for ~21 days. If I'm lucky, embryos will form, i grow those on a shaker in the dark for a week maybe. I grow them on solid agar media with a cocktail of hormones and MS media until they form a callus (a lumpy plant-tumor thing), and put it on MS with GA3 hormone and they will hopefully grow shoots, that I grow essentially as if it were a clone. What I'm doing is neat, I'm doing it on other species of brassica as well, stuff there aren't even methods papers on yet! Its essentially like "super-selfing," not sure what caveats would exist in a monoecious plant like Cannabis for something like this, but I would like to figure that out! It would dramatically reduce time needed to make true-breeding lines I would think, but I don't want to make baseless claims. Also, you do sometimes get cool mutants! for example, a mutation that caused the leaves on a botrytis (cauliflower) plant to wrap around the petiole like an elm leaf, and possibly a Siamese twin broccoli. here's a paper on this too! https://www.researchgate.net/publica...a_var_botrytis

Anther culture using pollen is a quick way to make a cannabis haploid. You have the aseptic protocol skills to do it... then use the haploids to make the double hap. 2 steps to pure homozygosity instead of 1 but still.
The beauty of cannabis is we can get a female to create pollen...

Tried multiple times with a shitty home made flow hood... not good enough.

You should totally fucking do this.

Two double haps crossed would make supreme f1s.

[Yard dog]

Originally Posted by zif

In contrast, if someone finds (and shares) an inducer male, you could pollinate a female and then simply sprout haploid seeds! In many species these actually show spontaneous chromosome doubling, so you have doubled haploids (I like to think of them as S∞) in one step. Who knows with cannabis? It may require treatment (e.g. with colchicine) to double....

I posted over in the recent findings thread that these genetics are getting worked out in maize. It’s quite possible that researchers with access to CRISPR or the like could intentionally make inducer males right now. It may well be that the mechanisms of pollen are sufficiently conserved across plant species.

Oddly, inducer males will probably seem horrible from a traditional breeding standpoint. They’re likely to produce seeds with low or super low germination. These ‘dud’ males may be the most valuable males anyone ever runs across! Crazy....

As far as I'm aware with regard to them using this with corn, they hit the selected mum with the super inducer to get her genes. They then double this up with treatment, the specific gene they termed "not like dad" is what they require for an inducer.

https://emboj.embopress.org/content/36/6/707