Industrial Hemp in Oregon

Sam_Skunkman said:

Good Luck,
I hope you are correct, mine all had lots of white nubs as well as set dark seeds, did you test them with only Triploid pollen or did you test them with Diploid pollen?
I only tested my 30 different unrelated Triploids (each plant over 6 feet tall 5 copies of each) with Diploid pollen from maybe a dozen unrelated Diploid Males, as that is the problem in the world, do you think that explains the difference? I know that in the world it is Diploid pollen they will run into and set seeds from. Maybe Triploid pollen is just not as viable? We had both THC and industrial hemp Triploids and Diploid Males we used to pollinate, they all set seed and all had even more white nubs than dark set seed. But the dark set seed was 10%-95% of normal yield of seeds, none had zero set seed, not including the white nub seeds found in all the Triploids.
Anyway, I am glad that 3 decades later this work is being pursued again, if it does lead to a useable breeding method, time will tell.
I did trials for 3 years before I gave up. But you know how R&D is most does not work out but you learn what does not work and occassionally do find R&D that moves Cannabis forward. Cannabis R&D is what I loved best besides smoking.
-SamS

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Hi Sam,

Happy to confirm that these triploids are definitely without seeds. Hsuan and team will be publishing later this fall, but it's very good news. Just cleaned a bucketload of material from different ploidy crossing attempts and there still isn't seed or nubs. Triploids pollinated by tetraploids, triploids, and diploids all yielded no seed. We are still waiting on the results for diploids pollinated by triploids to ensure that the pollen is truly sterile as well.

You guys paved the way on this and I know that our whole team is thankful for the advice and information. I'm grateful to share the same thrill of pursuit that R&D bring and it has been very cool to share this with a pioneer...most days are dead-ends but the good ones make waves that last! Wish you could have made it out this year, hopefully next summer.

Will post links when the formal papers are in press.

I really do not understand why all of the 30 different Triploid females set seed and all produced white nubbies maybe because we tried the pollination with many diploid males from different backrounds we had hemp males, Thai males, Afghan males, as well as hybrid males that we used. At least one of them did pollinate all the different varieties of 30 Triploid females, I had thought all the diploid males did make viable seeds in the triploids when we did the work. I am curious to see if tetraploid and triploid males pollen are viable to normal diploid females did you also test Tetraploid male pollen? Amazing work, it seems you found what I was looking for?

I agree about R&D most teaches what does not work but when you find something that does work all the R&D is worth it.

-SamS

socioecologist said:

Hi Sam,

Happy to confirm that these triploids are definitely without seeds. Hsuan and team will be publishing later this fall, but it's very good news. Just cleaned a bucketload of material from different ploidy crossing attempts and there still isn't seed or nubs. Triploids pollinated by tetraploids, triploids, and diploids all yielded no seed. We are still waiting on the results for diploids pollinated by triploids to ensure that the pollen is truly sterile as well.

You guys paved the way on this and I know that our whole team is thankful for the advice and information. I'm grateful to share the same thrill of pursuit that R&D bring and it has been very cool to share this with a pioneer...most days are dead-ends but the good ones make waves that last! Wish you could have made it out this year, hopefully next summer.

Will post links when the formal papers are in press.

Click to expand...

Man Sam I was just linked to this thread and I've got say I have the exact same concerns you do. Thank you for adding a voice of concerned interest in these projects from OCBD. Maybe everything is on the up and up but man, an entire catalog of sterile Triploid cultivars impervious to all pollen just doesn't sit well with me. My hope is I'm wrong and we'll never hear anything shady about OCBD but I'd be remiss if I didn't express some concerns and I appreciate your logical skepticism. Hopefully with time either grow reports, phylos testing or further explanations from OCBD will help illuminate the puzzle pieces I'm missing. Interesting project for sure, I'll be follow this whole development closely.

Sam_Skunkman said:

I really do not understand why all of the 30 different Triploid females set seed and all produced white nubbies maybe because we tried the pollination with many diploid males from different backrounds we had hemp males, Thai males, Afghan males, as well as hybrid males that we used. At least one of them did pollinate all the different varieties of 30 Triploid females, I had thought all the diploid males did make viable seeds in the triploids when we did the work. I am curious to see if tetraploid and triploid males pollen are viable to normal diploid females did you also test Tetraploid male pollen? Amazing work, it seems you found what I was looking for?

I agree about R&D most teaches what does not work but when you find something that does work all the R&D is worth it.

-SamS

Click to expand...

It's weird that they've managed to do what nobody else in cannabis has managed to do, ever. Kinda weird. Not sure if I'm buying it.

Sam, been thinking about this discrepancy and trying to understand it. How were the initial populations of tetraploids screened (flow cytometry, chromosome squash)? Were the triploids receiving pollen verified in the same way?

What we have seen with tetraploids is that reversion to the diploid state is common after the initial, successful doubling event. We have to take great care on the micropropagation side to prevent this from occurring, and it involves multiple "generations" of culture and/or cutting before relative stabilization. Plants that revert often end up mixoploid; some parts remain 4n while others revert to 2n; if that plant is used for seed or pollen production, some resulting seed would end up 3n, but others would be 2n and receptive to pollen.

One of the fun discoveries summer of 2019 was identifying spontaneous tetraploids in a population grown from seed. A larger question now is at what frequency those events happen (seems often!)--the answer of course will shape how we view the evolutionary history of the species.

rizraz said:

Man Sam I was just linked to this thread and I've got say I have the exact same concerns you do. Thank you for adding a voice of concerned interest in these projects from OCBD. Maybe everything is on the up and up but man, an entire catalog of sterile Triploid cultivars impervious to all pollen just doesn't sit well with me. My hope is I'm wrong and we'll never hear anything shady about OCBD but I'd be remiss if I didn't express some concerns and I appreciate your logical skepticism. Hopefully with time either grow reports, phylos testing or further explanations from OCBD will help illuminate the puzzle pieces I'm missing. Interesting project for sure, I'll be follow this whole development closely.

Click to expand...

Rest assured that diploid plants are not going away--ever--and that is not one of our goals. Pollen-proof cannabinoid production enables the peaceful coexistence fiber / grain production in areas most hospitable to cannabis production, while also benefitting from "gene stacking" via copy number increase of key genes of agronomic importance (disease resistance, secondary metabolite production, flower timing, etc.). While it's a certainly a big deal, it is not novel; genome doubling is the underlying tool used to make many of our most important crops "more fit" for agriculture.

I chuckled a bit at the mention of using Phylos to verify. A flow cytometer and seedlings is all you need for that. The ploidy of our seed will be independently verified in this way by Oregon State University's Seed Testing Laboratory as part of a standard germination trial.

I am not sure what's up with Phylos at the moment, as they have made no public statements in over a year and Mowgli Holmes has been demoted from CEO to a "strategic advisor". I do know, however, that their SNP-based "galaxy" is a really rudimentary comparison tool and has little to no application in ploidy verification. Their test accounts for ~2000 SNPs. There are many different competing genomics companies offering their own tests, ranging from 15K to 90K SNPS (the latter, Eurofins/Medicinal Genomics, was validated using one of our breeding populations as part of a collaboration with us). We are currently under contract with and testing other SNP platforms with Lighthouse Genomics (Canada) and NRGene (Israel) for novel trait detection.

SNP studies + phenotype data + chemotype data yield markers we can use to accelerate the breeding process for specific traits; however, whole genome data is also necessary to truly understand what is going on "under the hood" and to validate the usefulness of those candidate markers in other populations. We generate all that in house, utilizing PacBio's Sequel II HiFi sequencing and Bionano's optical sequencing to generate WGS for analysis. We have one of the largest--if not the largest--collections of WGS cannabis data in the world, all derived from our breeding projects. In addition to marker validation, these are being fed into a cannabis pangenome comparison tool to identify important structural variants, rates of translocation, and other areas of interest to our breeding team.

The same thing is going on in other crops right now because of the rapid evolution of sequencing technology; we got lucky that these tools became available at the same time "hemp" was legalized in the US. The rate of discovery is mind-blowing to say the least, which makes this thread such a fun piece of US cannabis history...you can see where we started and how it rapidly evolved.

Thanks for the reply. I removed some things from my post and made them a direct message. Below is just what was leftover so it may read a bit weird as a reply to you.

The white is an interesting project. I think some of the plants used to breed that plant may have been smuggled out of your facility because I was able to source a clone of the white about 4-6 months prior to public release.

Thanks for the replies, glad to hear sterile hemp isn't what you're after. Again this is a position you should clarify in my humble opinion because I think a lot of people (maybe a concerted effort) have been shared information that compares you all to Monsanto and if you're not super educated in Crawford history, the plant or the science, it makes a compelling argument.

<I am not sure what's up with Phylos at the moment, as they have made no public statements in over a year and Mowgli Holmes has been demoted from CEO to a "strategic advisor".>

He took I think $14 million dollars of investor money, and they want some sort of return. Good luck with that.

flylowgethigh said:

<I am not sure what's up with Phylos at the moment, as they have made no public statements in over a year and Mowgli Holmes has been demoted from CEO to a "strategic advisor".>

He took I think $14 million dollars of investor money, and they want some sort of return. Good luck with that.

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We used flow cytometry and found that you also need to check the roots, stems, leaves, flowers to be sure they are all Tetraploid or Triploid, we found many plants that were a mix of polyploids and diploids, the plants were aneuploids, or mixoploids. We did not use plants that were not 100% Tetraploids to make 100% Triploids for the female seed parents as I said before we used pollen parents that were only Diploids as that is what is found in nature that we were trying to prevent pollination from of our Triploid females.

We also found that tetraploids and Triploids can revert to Diploid. I am not sure if you can prevent this from occurring, by multiple "generations" of culture and/or cutting before relative stabilization. We just retested any Tetraploids or Triploids we wanted to use just before we used them, any plants with problems were thrown away.

We did not look for spontaneous Tetraploids in our normal Diploid populations so I can not talk about that or if they are stable or not.
I remember a paper that stated Cannabis landraces were found to be all Diploids. I can't remember the papers name or author.

socioecologist said:

Sam, been thinking about this discrepancy and trying to understand it. How were the initial populations of tetraploids screened (flow cytometry, chromosome squash)? Were the triploids receiving pollen verified in the same way?

What we have seen with tetraploids is that reversion to the diploid state is common after the initial, successful doubling event. We have to take great care on the micropropagation side to prevent this from occurring, and it involves multiple "generations" of culture and/or cutting before relative stabilization. Plants that revert often end up mixoploid; some parts remain 4n while others revert to 2n; if that plant is used for seed or pollen production, some resulting seed would end up 3n, but others would be 2n and receptive to pollen.

One of the fun discoveries summer of 2019 was identifying spontaneous tetraploids in a population grown from seed. A larger question now is at what frequency those events happen (seems often!)--the answer of course will shape how we view the evolutionary history of the species.

Click to expand...

rizraz said:

Thanks for the reply. I removed some things from my post and made them a direct message. Below is just what was leftover so it may read a bit weird as a reply to you.

The white is an interesting project. I think some of the plants used to breed that plant may have been smuggled out of your facility because I was able to source a clone of the white about 4-6 months prior to public release.

Thanks for the replies, glad to hear sterile hemp isn't what you're after. Again this is a position you should clarify in my humble opinion because I think a lot of people (maybe a concerted effort) have been shared information that compares you all to Monsanto and if you're not super educated in Crawford history, the plant or the science, it makes a compelling argument.

Click to expand...

The White CBG clone circulating and used by competitors to make seed in 2020 is an F1, not the production mother. The people managing our gene bank are entrusted, literally, with the collective livelihood of everyone else in our company and their trust is beyond reproach. We know at least 1 of the 32 farms that conducted 2019 field trials with this variety was selling F1 starts in June / July of that year. Could be more--doesn't really matter at this point. Our 2019 trials were in Oregon, California, Nevada, South Carolina, and Vermont. The phenotype, chemotype, and genotype segregation in other companies' CBG varieties this year show, without a doubt, that they are F2s (we've grown / tested a bunch to confirm this).

We entrusted our national field trials to a network of land grant universities this year (17 locations), run by Oregon State University through a USDA-backed hemp research study. Hopefully it produces less scurrilous results.

RE: Monsanto. This year has really brought the world into focus. Humanity seems more divided than ever, fomented by purposeful disinformation meant only to gather attention, which in itself has been monetized (i.e. see the "Social Dilemma"). I see this similarly. If someone yells "Monsanto" at us, they can consistently get 25% of the audience to chime in and yell "yea! fuck monsanto!", then two months later they are slinging seeds to the gullible farmers based on an emotional delusion. Year after year after year. It works, because for most, information < emotion.

Monsanto ceased being a company in 2018. It's just "Bayer AG" now, with Monsanto bought for $63 billion. Oregon CBD <----> Bayer comparison? Bayer had $43 BILLION in revenue in 2019, employed over 103,000 people, and is a critical member of a global oligopoly in seed / fertilizer / pesticides. They are publicly traded and have a total market capitalization of over $66 billion. They also have a long, dark history that includes using slave labor at concentration camps during WWII (i.e. genocide) and the widespread adulteration of our planet with glyphosate, among other non-trivial things.

Oregon CBD is an LLC owned by Seth and Eric Crawford (no outside investors, ever) that employs 35 people. 2020 revenue is negative, with gross receipts down 300% from 2019. By employment numbers, we were ranked by the Portland Business Journal as the 9th largest cannabis business...in Oregon. We are not even ranked in the top-10 largest businesses in our rural Polk county. We are a TINY company trying to do big things in a space we love, while holding on for dear life during the most turbulent time we've ever experienced. We don't support genocide, slavery, or glyphosate.

Sam_Skunkman said:

We used flow cytometry and found that you also need to check the roots, stems, leaves, flowers to be sure they are all Tetraploid or Triploid, we found many plants that were a mix of polyploids and diploids, the plants were aneuploids, or mixoploids. We did not use plants that were not 100% Tetraploids to make 100% Triploids for the female seed parents as I said before we used pollen parents that were only Diploids as that is what is found in nature that we were trying to prevent pollination from of our Triploid females.

We also found that tetraploids and Triploids can revert to Diploid. I am not sure if you can prevent this from occurring, by multiple "generations" of culture and/or cutting before relative stabilization. We just retested any Tetraploids or Triploids we wanted to use just before we used them, any plants with problems were thrown away.

We did not look for spontaneous Tetraploids in our normal Diploid populations so I can not talk about that or if they are stable or not.
I remember a paper that stated Cannabis landraces were found to be all Diploids. I can't remember the papers name or author.

Click to expand...

Exactly right on the re-test re-test re-test, toss if there's any doubt. Got to say that this has me perplexed. I'll continue to update as we learn more, but I understand your doubt having done everything exactly right and getting a different outcome.

A member posted this in the "triploid" thread; I had not read it before. A natural, isolated tetraploid population was found in Darcha, India. High (3,360m), dry, and cold. https://www.eresearchco.com/jbcc-pdf/6%20VKS%20JBCC%202_1_.pdf

Hey Seth, Thanks for the reply. On the CBG I received a clone of the white in Vermont way before any seeds could have been produced, unsure the origins. I'm happy to send a clone to you for examination. Just wanted to let you know that as best as I can tell I got a plant that wasn't released yet long before it could have been remade. The CBG plant I have, I only grow for the production of compliant Rosin. I haven't distributed it or reproduced it. I did see a farm working this year's release of the white and I will say mine and his are very closely related. I haven't pursed any more CBG strains personally because to be honest I don't like the smoke. Something about the fragile structure of the trichomes and how easily they separate from the flower gives me the sneezes. It does make stunning bright white rosin with zero percent THC so that's something.

Yeah I hate that as well regarding Monsanto name calling. It seems like we've entered a stage of debating/questioning where the point isn't to uncover information but rather insight crowd mentality against a point. I want to be clear here, I know you're nothing like Monsanto. I'm not sure any Cannabis seed producer could even dream of getting that big. I also am fully aware that you're privately owned and I'm lucky enough to remember you all being around back in day at a few of the Cannabis conventions I got dragged to. I think what you said here "information < emotion" sums it up perfectly and thank you for this quote as well "We don't support genocide, slavery, or glyphosate." Thanks for the replies Seth.

socioecologist said:

The White CBG clone circulating and used by competitors to make seed in 2020 is an F1, not the production mother. The people managing our gene bank are entrusted, literally, with the collective livelihood of everyone else in our company and their trust is beyond reproach. We know at least 1 of the 32 farms that conducted 2019 field trials with this variety was selling F1 starts in June / July of that year. Could be more--doesn't really matter at this point. Our 2019 trials were in Oregon, California, Nevada, South Carolina, and Vermont. The phenotype, chemotype, and genotype segregation in other companies' CBG varieties this year show, without a doubt, that they are F2s (we've grown / tested a bunch to confirm this).

We entrusted our national field trials to a network of land grant universities this year (17 locations), run by Oregon State University through a USDA-backed hemp research study. Hopefully it produces less scurrilous results.

RE: Monsanto. This year has really brought the world into focus. Humanity seems more divided than ever, fomented by purposeful disinformation meant only to gather attention, which in itself has been monetized (i.e. see the "Social Dilemma"). I see this similarly. If someone yells "Monsanto" at us, they can consistently get 25% of the audience to chime in and yell "yea! fuck monsanto!", then two months later they are slinging seeds to the gullible farmers based on an emotional delusion. Year after year after year. It works, because for most, information < emotion.

Monsanto ceased being a company in 2018. It's just "Bayer AG" now, with Monsanto bought for $63 billion. Oregon CBD <----> Bayer comparison? Bayer had $43 BILLION in revenue in 2019, employed over 103,000 people, and is a critical member of a global oligopoly in seed / fertilizer / pesticides. They are publicly traded and have a total market capitalization of over $66 billion. They also have a long, dark history that includes using slave labor at concentration camps during WWII (i.e. genocide) and the widespread adulteration of our planet with glyphosate, among other non-trivial things.

Oregon CBD is an LLC owned by Seth and Eric Crawford (no outside investors, ever) that employs 35 people. 2020 revenue is negative, with gross receipts down 300% from 2019. By employment numbers, we were ranked by the Portland Business Journal as the 9th largest cannabis business...in Oregon. We are not even ranked in the top-10 largest businesses in our rural Polk county. We are a TINY company trying to do big things in a space we love, while holding on for dear life during the most turbulent time we've ever experienced. We don't support genocide, slavery, or glyphosate.

Click to expand...

That plant they found in the Triploid thread I started was maybe one of the more interesting things I've read about in a long while. I share Sam's concerns about reversions back to Diploids. I wonder if you can stop that natural progression back to the original state? I also have read a bunch of stuff that had the exact same results that Sam posted. Seems like most of these project end in some aneuploids or mixoploids plants. I guess time will tell if you've escaped those pitfalls. I'll be following along closely and thanks for replies.

socioecologist said:

Exactly right on the re-test re-test re-test, toss if there's any doubt. Got to say that this has me perplexed. I'll continue to update as we learn more, but I understand your doubt having done everything exactly right and getting a different outcome.

A member posted this in the "triploid" thread; I had not read it before. A natural, isolated tetraploid population was found in Darcha, India. High (3,360m), dry, and cold. https://www.eresearchco.com/jbcc-pdf/6%20VKS%20JBCC%202_1_.pdf

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Not sure if anyone knows the answer to this

Has there been any success lately in breeding fiber/seed strains of hemp that will thrive commercially in a tropical climate?

I looked through this thread and it seemed to me that most hemp was grown in more temperate zones and that most varietals did best in longer day periods and would flower too soon if grown with under 13 hours of light a day.

Thanks in advance for any insight!

Hopefully I can get a "pass" for not providing many updates or details here, but it's been 2020 (a noun and adjective) all year. The year of virus, crushing regulations, social disorder, and widespread climatic extremes, to name only a few. We're over this year and looking forward to the upside of down. The year captured in a picture, taken during Oregon's worst smoke inundation event of memory, which lasted 16 days during the first two weeks of September. It's felt like farming in the apocalypse and, in many ways, has been.

With the winter solstice quickly approaching, Nature reminds us that it's time to prepare for a new season of growth. What will 2021 hold? We've given up trying to predict this upside-down world and, instead, are focusing on what we can contribute to the collective shaping of a brighter future for all. The "Up Side of Down," if you will. Sometimes a simple shift in consciousness is all it takes.

2021 will mark our 7th year in hemp operations and our 6th season of providing the industry's leading feminized seed to farmers. For 2021, all of our seed is certified both organic and non-GMO. Attaining these important benchmarks provides an even higher level of trust in our production processes and more value to farmers in the commodity chain. Factor in the industry's leading feminization rates, best total cannabinoid yields per acre, a bevy of prestigious awards for our genetics, and the revolutionary "seedless" triploid breeding advancements we are introducing, and we believe farmers' choice of seed suppliers has become more clear than ever before.

Our whole team constantly strives to be more efficient, make even better varieties, and to give back wholeheartedly, especially in the face of catastrophe and challenges. The recent installation of 1.1 megawatts of solar generation capacity (2 acres of panels) at our main campus is our first step towards adding "net-zero" to the list of positive reasons to support Oregon CBD. More importantly, it provides a tangible manifestation of positive change and shows that hope is never lost--it emerges within those who choose to cultivate it.

Very inspirational.
I remember watching you start this thread. Coming from sea cans and a garage to this in only 6 years is remarkable. You and your team deserve a pat on the back. Well done.
I hope I can achieve a fraction of this up here in Canada on a similar path.

Peace GG

socioecologist said:

With the winter solstice quickly approaching, Nature reminds us that it's time to prepare for a new season of growth. What will 2021 hold? We've given up trying to predict this upside-down world and, instead, are focusing on what we can contribute to the collective shaping of a brighter future for all. The "Up Side of Down," if you will. Sometimes a simple shift in consciousness is all it takes.

2021 will mark our 7th year in hemp operations and our 6th season of providing the industry's leading feminized seed to farmers. For 2021, all of our seed is certified both organic and non-GMO. Attaining these important benchmarks provides an even higher level of trust in our production processes and more value to farmers in the commodity chain. Factor in the industry's leading feminization rates, best total cannabinoid yields per acre, a bevy of prestigious awards for our genetics, and the revolutionary "seedless" triploid breeding advancements we are introducing, and we believe farmers' choice of seed suppliers has become more clear than ever before.

Our whole team constantly strives to be more efficient, make even better varieties, and to give back wholeheartedly, especially in the face of catastrophe and challenges. The recent installation of 1.1 megawatts of solar generation capacity (2 acres of panels) at our main campus is our first step towards adding "net-zero" to the list of positive reasons to support Oregon CBD. More importantly, it provides a tangible manifestation of positive change and shows that hope is never lost--it emerges within those who choose to cultivate it.

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Wow - awesome solar farm!