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Old 02-22-2018, 08:55 PM #461
Sam_Skunkman
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Originally Posted by socioecologist View Post
Well shit, no arrogance intended and apologies to everyone if it came across that way. Thanks for the kind words and redirect if I was out of line. Sam has always provided the best advice in terms of searching for something unique (grow as many plants as possible), so the end of the post was supposed to reflect that sage, but often overlooked wisdom.

I thought Mate Dave was asking about the youPCR picture that Sam posted and how that would / could translate to breeding. We've been working with the platform since last summer. Screening for male / female is robust, accurate, fast, relatively cheap. Screening for active CBDa or THCa alleles is much more difficult. Kevin McKernan @ Medicinal Genomics is always working to make the test better by adding new active allele data as it comes in, but the test is very temperature sensitive and we are still getting occasional false positives on the CBDa side. We use the youPCR screen to segregate type I / type IV and type III / type IV in our current projects.

The platform works great for type I / II / III segregation that many people are working on to breed better type IIs and IIIs. When a pure THC parent (P1) is crossed with a pure CBD parent (P2) and the resulting progeny open pollinated / selfed, the square for F2s looks like this (generated from the previously linked website) :

T = functional THCa allele
t = nonfunctional THCa allele
D = fuctional CBDa allele
d = nonfunctional CBDa allele

Which of the 9 types groups had the highest total Cannabinoids and which group had the highest THC only? CBD? CBG?
I am wondering if the Cannabinoid genes or the genes that allow high expression, lots of big resin heads for example are the most important.
When you say functional THCa allele you are referring to the THC synthase correct?

-SamS

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Old 02-22-2018, 09:04 PM #462
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Originally Posted by socioecologist View Post
Thanks Sam. 12 seeds to the gram??? That's incredible. Our largest is 40 seeds per gram and I have a hard time imagining how it could get any bigger. Very cool.

Day neutral are definitely lower in total cannabinoid content, but not prohibitively so. We have them averaging 12% CBD in our type III field varieties and 10% CBG in our type IVs. Their photoperiod counterparts have about 20% higher content on average (15% and 12%, respectively). (note: all tests performed by NELAP accredited labs).

Two other things to think about when considering the relative efficiency of day neutral vs. photoperiod: (1) stem weight and (2) multiple harvests. We are finding that the average stem weight (as a proportion of total plant weight) on full season photoperiod varieties is about 27%; day neutrals come in at 18%. Autos allow at least two harvests per season in a decent production environment or five if you plan to grow in the central valley of California...

Having plants that can reliably finish in late July / early August allows certain locations (i.e. Oregon's Willamette valley) to field dry; when costly drying infrastructure is removed from the equation and finished plants can be picked up with a combine, day neutrals start to look pretty appealing.

Here's one of our berry flavored type III autos; they have more resin than many photoperiod varietals.
Is the plant seeded? Looks like it. I do agree this auto does have resin. Does it finish flowering and frost out more like normal drug Cannabis? That was the problem with Autos 25 years ago they just kept on making flowering biomass and never really stopped making flowers and frosting out. `if you test the DNA of any CBD Autos it would be interesting to see if they contained FINOLA DNA as I suspect, even if not it would be interesting to understand the relationships in the AUTO varieties.

-SamS

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Old 02-23-2018, 06:59 PM #463
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Originally Posted by Sam_Skunkman View Post
Is the plant seeded? Looks like it. I do agree this auto does have resin. Does it finish flowering and frost out more like normal drug Cannabis? That was the problem with Autos 25 years ago they just kept on making flowering biomass and never really stopped making flowers and frosting out. `if you test the DNA of any CBD Autos it would be interesting to see if they contained FINOLA DNA as I suspect, even if not it would be interesting to understand the relationships in the AUTO varieties.

-SamS
It was indeed seeded; F1 in the picture, now in the F4. Your description of their growth is spot on, though they continue to produce copious resin even while building more and more flowers. Great oil producers with complex astringent berry flavors on this one. I don't know for sure, but I suspect all of the autos on the market today were originally derived from Lowryder. The bigger question is what that particular plant (Lowryder) was derived from. I doubt it is Finola; when I grew the original Lowryder and Lowryder II autos in the mid-2000s, it performed nothing like the Finola we experimented with the last few years.

Can the P1 and P2 of Finola remain in veg state under 24 hours of light? Or am I making an incorrect assumption about Jace's seed making strategy?

One of our ongoing research projects is to locate and better understand the genes affecting flowering in cannabis. Plants like these (autos, Finola, and hybrids) are all helping us get a better handle on what's going on under the hood.

RE: the Punnet square. That was just for reference on F2 genotype segregation for the person asking about it. You are correct that certain genotypes tend to produce higher levels of cannabinoids. Homozygous > Heterozygous for pure types (I, III), with heterozygous type IIs producing the most overall.
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Old 02-25-2018, 08:23 PM #464
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The genetics of haploid inducers in maize are becoming clearer (see below). I wonder how far away an inducer male is for the cannabis community? Or is he already out there somewhere?

If so, I wonder what his effects would look like? Would he cause females to have a normal appearing seed set, but with a very high percentage of those seeds nonviable?

Gilles et al. (2017) Loss of pollen-specific phospholipase NOT LIKE DAD triggers gynogenesis in maize. doi:10.15252/embj.201796603
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Old 02-26-2018, 10:24 AM #465
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they should teach genetics in high school with marijuana as an example. I think students would learn a lot faster.
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Old 02-26-2018, 11:19 AM #466
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they should teach genetics in high school with marijuana as an example. I think students would learn a lot faster.
I'm going way back here. In the mid 70s, a couple friends and I were smoking a joint outside a door in grade 12 and my math teacher came out and caught us. All he did was take it, took a good haul, and handed it back. "don't tell anyone guys. Go around the corner to finish it". He was the only teacher with " long hair". Lol

Great teacher BTW. His classes were fun (for a math class)..
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Old 02-26-2018, 06:28 PM #467
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https://emboj.embopress.org/content/e...embj.201796603

-SamS



Quote:
Originally Posted by zif View Post
The genetics of haploid inducers in maize are becoming clearer (see below). I wonder how far away an inducer male is for the cannabis community? Or is he already out there somewhere?

If so, I wonder what his effects would look like? Would he cause females to have a normal appearing seed set, but with a very high percentage of those seeds nonviable?

Gilles et al. (2017) Loss of pollen-specific phospholipase NOT LIKE DAD triggers gynogenesis in maize. doi:10.15252/embj.201796603
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Old 04-07-2018, 05:24 PM #468
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Subcellular localization defines modification and production of Δ9-tetrahydrocannabinolic acid synthase in transiently transformed Nicotiana benthamiana.

https://www.ncbi.nlm.nih.gov/pubmed/29619743

CONCLUSION:
Nicotiana benthamiana is a suitable host for the generation of cannabinoid producing enzymes. To attain whole pathway integration, careful analysis of subcellular localization is necessary.
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Old 04-08-2018, 03:29 PM #469
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Originally Posted by bsgospel View Post
CONCLUSION:
Nicotiana benthamiana is a suitable host for the generation of cannabinoid producing enzymes. To attain whole pathway integration, careful analysis of subcellular localization is necessary.
Thanks for sharing this. Insertion of THCa synthase into tobacco = successful. Note that this was not the insertion of the full cannabis chemical generation apparatus, just THCa. To produce THCa, they had to place the altered isolated tobacco plant cells in a CBGa rich substrate (i.e. the plant doesn't make precursors natively).

The additional resulting compounds produced are, to me, the really interesting part of this study. The last paragraph of the paper notes:

Quote:
"THCAS does convert CBGA to different products in a plant matrix, suggesting that enzyme promiscuity additionally contributes to the diversity of cannabinoids in Cannabis."
That particular THCa synthase, in vitro, produces CBCa as well (albeit 4x less affinity). Know your synthases!
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