[socioecologist]

Originally Posted by Mate Dave

Can you elaborate on how we can use this Sam? A little guidance perhaps as to how we would be able to predict the punnetts. Would we be punnetting the Chemotypes? How?

First, know what your end goals are and how to get there. Read the papers posted by Sam. That will take a bit. A helpful tool for squares (remove the "s" in https):

scienceprimer.com/punnett-square-calculator

Then, buy PCR and DNA extraction gear. Learn how to use it. Run hundreds of tests. Alter the protocol to dial in accurate prediction of your particular alleles with secondary chemical analysis (temperature gradient testing with known material is key here). Voila! You have yourself a basic chemotyping tool via PCR.

View image in gallery

Oh, and grow some plants. Hundreds of thousands per year at minimum.

[led05]

Originally Posted by socioecologist

First, know what your end goals are and how to get there. Read the papers posted by Sam. That will take a bit. A helpful tool for squares (remove the "s" in https):

scienceprimer.com/punnett-square-calculator

Then, buy PCR and DNA extraction gear. Learn how to use it. Run hundreds of tests. Alter the protocol to dial in accurate prediction of your particular alleles with secondary chemical analysis (temperature gradient testing with known material is key here). Voila! You have yourself a basic chemotyping tool via PCR.

View Image
View image in gallery

Oh, and grow some plants. Hundreds of thousands per year at minimum.

You brothers are amazing, what you're doing great but this post in a way strikes me as arrogant, unlike any other you've ever done...

don't get too big for them britches and remember your roots

Thanks for your work

if I had to guess, offers or $$ flow is a coming, stay true

[socioecologist]

Originally Posted by led05

You brothers are amazing, what you're doing great but this post in a way strikes me as arrogant, unlike any other you've ever done...

don't get too big for them britches and remember your roots

Thanks for your work

if I had to guess, offers or $$ flow is a coming, stay true

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

View image in gallery

[led05]

Originally Posted by socioecologist

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

View Image
View image in gallery

Like I said...- amazing, you guys..... it seems this industry corrupts the best; probably worst too but whatever; I've watched you/team posts for a long time and have always been amazed by the humility and truthiness that resonates, on other forums too, I believe you've always had same name?

Anyhow, not a dick-rub; excuse me but hope others will follow.... It's so inevitable that big $ will swamp this space and run it; run it to the extent small or mid money that is involved and believes is washed over....

I wonder how Peter C feels in Co lately,this is why CO is so special, the folks bucked the big $ directly, it's truly special and in history will be way more significant than realized today.

[socioecologist]

Legendary plant biologist Dr. Ernest Small released the most comprehensive agronomy and breeding paper on cannabis to date (“Dwarf germplasm: the key to giant Cannabis hempseed and cannabinoid crops”). He calls for the development of cannabinoid-rich, large seeded, day neutral varieties and hails them as the future of cannabis production, while lamenting research restrictions in place preventing the use of appropriate germplasm (i.e. high resin content type I) to develop these varieties.

Full article here: sci-hub.cc/10.1007/s10722-017-0597-y

*** Abstract ***

After a century of banishment, both euphoric (‘‘marijuana’’) and non-euphoric (‘‘industrial hemp’’) classes of Cannabis sativa are attracting billions of dollars of investment as new legitimate crops. Most domesticated C. sativa is very tall, a phenotype that is desirable only for hemp fibre obtained from the stems. However, because the principal demands today are for chemicals from the inflorescence and oilseeds from the infructescence, an architecture maximizing reproductive tissues while minimizing stems is appropriate. Such a design was the basis of the greatest short-term increases in crop productivity in the history of agriculture: the creation of short-stature (‘‘semi-dwarf’’), high-harvest-index grain cultivars, especially by ideotype breeding, as demonstrated during the ‘‘Green Revolution.’’ This paradigm has considerable promise for C. sativa. The most critical dwarfing character for breeding such productivity into C. sativa is contraction of internodes. This reduces stem tissues (essentially a waste product except for fibre hemp) and results in compact inflo- rescences (which, on an area basis, maximize cannabi- noid chemicals) and infructescences (which maximize oilseed production), as well as contributing to ease of harvesting and efficiency of production on an area basis. Four sources of germplasm useful for breeding semi-dwarf biotypes deserve special attention: (1) Naturally short northern Eurasian wild plants (often photoperiodically day-neutral, unlike like most bio- types) adapted to the stress of very short seasons by maximizing relative development of reproductive tissues. (2) Short, high-harvest-index, oilseed plants selected in northern regions of Eurasia. (3) ‘‘Indica type’’ marijuana, an ancient semi-dwarf cultigen tracing to the Afghanistan-Pakistan area. (4) Semi- dwarf strains of marijuana bred illegally in recent decades to avoid detection when grown clandestinely indoors for the black market. Although the high THC content in marijuana strains limits their usage as germplasm for low-THC cultivars, modern breeding techniques can control this variable. The current elimination of all marijuana germplasm from breeding of hemp cultivars is short-sighted because marijuana biotypes possess a particularly wide range of genes. There is an urgent need to develop public gene bank collections of Cannabis.

[Sam_Skunkman]

If the link does not work try sci-hub.la/

Dwarf germplasm: the key to giant Cannabis hempseed and cannabinoid crops

Interesting paper I was sent last month. That said I found some I disagreed with, I sent him a response, explaining why. Nice reply from Ernest.

Auto day neutral varieties do not normally produce as much resin, that is a problem for the other Cannabinoids if they are the targets.

I have known Ernie for years and like his work.

BTW I have small seeds 1.25 grams to 1000 seeds, 800+ to the gram as well as big seeds that are 83 grams for 1000 seeds, 12 to the gram, that is a wider range then he reported.
-SamS

Originally Posted by socioecologist

Legendary plant biologist Dr. Ernest Small released the most comprehensive agronomy and breeding paper on cannabis to date (“Dwarf germplasm: the key to giant Cannabis hempseed and cannabinoid crops”). He calls for the development of cannabinoid-rich, large seeded, day neutral varieties and hails them as the future of cannabis production, while lamenting research restrictions in place preventing the use of appropriate germplasm (i.e. high resin content type I) to develop these varieties.

Full article here: sci-hub.cc/10.1007/s10722-017-0597-y

*** Abstract ***

After a century of banishment, both euphoric (‘‘marijuana’’) and non-euphoric (‘‘industrial hemp’’) classes of Cannabis sativa are attracting billions of dollars of investment as new legitimate crops. Most domesticated C. sativa is very tall, a phenotype that is desirable only for hemp fibre obtained from the stems. However, because the principal demands today are for chemicals from the inflorescence and oilseeds from the infructescence, an architecture maximizing reproductive tissues while minimizing stems is appropriate. Such a design was the basis of the greatest short-term increases in crop productivity in the history of agriculture: the creation of short-stature (‘‘semi-dwarf’’), high-harvest-index grain cultivars, especially by ideotype breeding, as demonstrated during the ‘‘Green Revolution.’’ This paradigm has considerable promise for C. sativa. The most critical dwarfing character for breeding such productivity into C. sativa is contraction of internodes. This reduces stem tissues (essentially a waste product except for fibre hemp) and results in compact inflo- rescences (which, on an area basis, maximize cannabi- noid chemicals) and infructescences (which maximize oilseed production), as well as contributing to ease of harvesting and efficiency of production on an area basis. Four sources of germplasm useful for breeding semi-dwarf biotypes deserve special attention: (1) Naturally short northern Eurasian wild plants (often photoperiodically day-neutral, unlike like most bio- types) adapted to the stress of very short seasons by maximizing relative development of reproductive tissues. (2) Short, high-harvest-index, oilseed plants selected in northern regions of Eurasia. (3) ‘‘Indica type’’ marijuana, an ancient semi-dwarf cultigen tracing to the Afghanistan-Pakistan area. (4) Semi- dwarf strains of marijuana bred illegally in recent decades to avoid detection when grown clandestinely indoors for the black market. Although the high THC content in marijuana strains limits their usage as germplasm for low-THC cultivars, modern breeding techniques can control this variable. The current elimination of all marijuana germplasm from breeding of hemp cultivars is short-sighted because marijuana biotypes possess a particularly wide range of genes. There is an urgent need to develop public gene bank collections of Cannabis.

[socioecologist]

Originally Posted by Sam_Skunkman

If the link does not work try sci-hub.la/

Interesting paper I was sent last month. That said I found some I disagreed with, I sent him a response, explaining why. No answer from Ernest yet. Auto day neutral varieties do not normally produce as much resin, that is a problem for the other Cannabinoids if they are the targets.
I have known Ernie for years and like his work.
BTW I have small seeds 1.25 grams to 1000 seeds, 800+ to the gram as well as big seeds that are 83 grams for 1000 seeds, 12 to the gram, that is a wider range then he reported.
-SamS

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.

[Sam_Skunkman]

Do you also grow hemp for fiber? IF SO HOW RETTED? We found field retted in E Europe and China, easy but not as good quality as water retted for fiber. Specially color was better, with water retted.
I found several others that have seeds 30 to the gram. But 800+ to the gram never have I seen anyone close.
-SamS

Originally Posted by socioecologist

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.

View Image

[socioecologist]

Originally Posted by Sam_Skunkman

Do you also grow hemp for fiber? IF SO HOW RETTED? We found field retted in E Europe and China, easy but not as good quality as water retted for fiber. Specially color was better, with water retted.
I found several others that have seeds 30 to the gram. But 800+ to the gram never have I seen anyone close.
-SamS

Fiber is a byproduct for us (animal bedding), so I have little experience there.

800 seeds to the gram is preposterously small. We have some varieties with "small" seeds (0.7g / 100), but nothing even remotely close. Did it pose germination issues or affect overall vigor? Our smallest seed variety produces the highest average cannabinoid content of all our lines.

[Sam_Skunkman]

This was a wild landrace variety I collected 9000 feet in Kashmir, the THC is very low and the seeds sprout very easy, seed production is very easy, seed harvest and collection is very hard as when you touch the plants the slightest the mature seeds seem to be spring loaded and are propelled from the plant. I collected the seeds very carefully with an upside down umbrella with a slit to allow the umbrella under the plant, that worked fairly good.
I suspect they are so easily shattering because they are very wild and man has not bred them or selected them for what man wants, bigger seeds and non shattering seeds in the flowers.
I did grow them at sea level in Santa Cruz to see if the would effect seeds size, it did not they were just as small.
-SamS