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LONG TERM SEED STORAGE 20+ YEARS
- Thread starter Sam_Skunkman
- Start date
XSir, we have got you covered.
Thanks for all of the information you have shared over the last decade. Others are picking up the torch where you have left it. It has been an incredible inspiration knowing you, and seeing what you have accomplished even under such a strict prohibition. The dedication you have shown the species is incredible and largely unknown, and I thank you for the efforts you have put forth in the last 50 years.
Re: single terps vars. - that is highly unlikely in my opinion. Single terpene synthases are able to generate multiple possible terpenic outcomes. We're starting to see the different possible combinations and ratios, depending on which terps you qualify/quantify; however it seems to fall mostly into classes rather than single terpene profiles as it does with cannabs.
That said, I still think people should try. I understand the problems with single terpene varieties, both about the synthases and/or possible linkages, but even if we can only create some single terpene varieties like the ones I have seen with only myrcene, or others, that is a start in the right direction.
One thing I can say is that terpene profiles are inheritable and that implys you can tailor the profiles and maybe even create single terpene varieties for each terpene, it just needs to be tried with more of the terpenes.
Seen this Chimera?
Expression and characterization of terpene synthases
from Cannabis sativa L. and Salvia sclarea L.
www.researchgate.net/...terpene_synthases.../72e7e5269a0dfd0663.pdf
I can't get the pdf file to be complete just add http://www. and .pdf before and at the end, of the below it works....
researchgate.net/...terpene_synthases.../72e7e5269a0dfd0663
Or paste this into google and the link will pop up.
And this?
The nonmevalonate pathway supports both monoterpene and sesquiterpene formation in snapdragon flowers
Natalia Dudareva, Susanna Andersson, Irina Orlova, Nathalie Gatto, Michael Reichelt, David Rhodes, Wilhelm Boland, Jonathan Gershenzon
Proc Natl Acad Sci U S A. Jan 18, 2005; 102(3): 933–938.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC545543/
I hope you do lots of terpene research!
-SamS
I look forward to the moment when we can publicly share the research we've been looking at over the past few years.
Regardless, your life's work has been educational to me personally, and I thank you for your contributions. Times spent with yourself and discussions with you and the Mrs have been incredibly educational and eye opening. I look forward to you having a chance for you to visit some day and hope we can offer the same in kind.
Respectfully,
-Chimera
PS- I would be interested if you could do a TTC assay on the non-germinating seeds to determine if they are actually dead, or rather just dormant...
I don't think I will have time to test the dormant/dead seeds, I do not have what I need at hand. It would be easy to repeat the germination testing with the worst germinating seeds after I get my tetrazolium chloride, I will try.
-SamS
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Hi Sam and Chimera
Hey that is an interesting topic. I am interested in it too. I thought about making a post centered on the concept using this paper as a starting point:
Glandular trichomes: what comes after expressed sequence tags?
This is another gem of a paper, it's not specifically about Cannabis, but a lot of the core concepts would carry over.
I find it fascinating to watch as our understanding of the forces that determine form and behavior in organisms are elucidated, and our simplistic, old paradigms develop into a truer understanding, a more powerful understanding. While acknowledging the incredible success of the gene centered Mendelian mindset, the modern thinker recognizes that the regulation of the expression of these genes is just as important. New techniques like EST allow us to probe the machinary of gene expression. As our understanding of the transcriptome and epigenome etc grows, our command of the form of an organism will become truly breath taking. I also believe that the mechanisms of cytoplasmic inheritance will take a more prominent spot in our attention. Doesn't it seem that some progress could be made in being able to breed for organelle quality? Super mitochondria and chloroplasts? I guess we'll see.
Anyway, the above paper is a real gold mine, just the references alone could keep you busy for a long time.
As a side note, the method they mention for isolating the trichomes mirrors an approach I came up with some years ago that I have been slowly developing. Check this out:
If you use very dry material, over a sieve, cold, in a matrix of silica gel beads, with ultrasonic deblinding, you get a surprisingly good quality at an excellent rate of return.
As I am sure you guys are painfully aware, lab equipment prices are beyond insane, so I DIY my own stuff when possible, as in this case. I have learned a lot about sifting and ultrasonics, and my prototype test beds have given me more data to work with than I can handle at the moment. I can tell you that if you load the top deck of a gyratory sifter (w/ultrasonic deblinding and the right mesh sizes) with really dry, frosty material, activated silica gel beads, and small dry ice pieces, you get some very interesting results indeed.
EDIT: I did not build the unit in the picture, lol! That is a commercial unit for lab use that I just used for illustration as it has a good view of the ultrasonic deblinding unit.
Attachments
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Nice thread, subbed.
Understated, so, Holy shitsnacks!
I feel something special may come from this,
who's with me?
Understated, so, Holy shitsnacks!
I feel something special may come from this,
who's with me?
S
sirius haze
seed batch 153 seems not to be pure NLD, i suspect some WLD in it 
Hi Sam,
Going back to one of your first questions about the why do or don't they germinate:
One factor is the abscisic acid (ABA) content in the seeds which keeps them dormant (in a 'sleeping beauty state'). It can 'wear off' over time and hence the seeds try to come alive even at 4°C thereby using up energy and other resources which ultimately lack the day you try to germinate them years later. On the other hand, it can also be too concentrated leading to seeds that need better washing or just longer in the wet towel or soil. That's those seeds where the ABA antagonist gibberellin (e.g. GA3) increases germination rates. You surely know that GA3 is widely used to boost germination rates in older seeds
.
A second factor might be seed shell thickness. Fine shells might protect less...
And to the single terpene plants: Myrcene is somewhat an exception because it is a precursor for many other terpenes. If you have a variety which lacks most terpene synthases, myrcene accumulates. It's a bit like cannabigerol acid which increases when you lack the THCA/CBDA synthases .
Going back to one of your first questions about the why do or don't they germinate:
One factor is the abscisic acid (ABA) content in the seeds which keeps them dormant (in a 'sleeping beauty state'). It can 'wear off' over time and hence the seeds try to come alive even at 4°C thereby using up energy and other resources which ultimately lack the day you try to germinate them years later. On the other hand, it can also be too concentrated leading to seeds that need better washing or just longer in the wet towel or soil. That's those seeds where the ABA antagonist gibberellin (e.g. GA3) increases germination rates. You surely know that GA3 is widely used to boost germination rates in older seeds
.A second factor might be seed shell thickness. Fine shells might protect less...
And to the single terpene plants: Myrcene is somewhat an exception because it is a precursor for many other terpenes. If you have a variety which lacks most terpene synthases, myrcene accumulates. It's a bit like cannabigerol acid which increases when you lack the THCA/CBDA synthases
.
seed batch 153 seems not to be pure NLD, i suspect some WLD in it
Why do you say that, from the WLD cotyledons?
-SamS
Hi OO, I thought you would comment in this thread.
Here are a couple more papers that illustrate the genome+transcriptome+epigenome etc multivariate "wider canvas" new paradigm that are germane to the OP.
From Kew, not a research paper, but a note on one of their projects:
Plant Dormancy Transcriptomes and Epigenetic Control
Research paper:
Arabidopsis Paired Amphipathic Helix Proteins SNL1 and SNL2 Redundantly Regulate Primary Seed Dormancy via Abscisic Acid–Ethylene Antagonism Mediated by Histone Deacetylation
Research paper with a broader view of the same mechanisms in plants in general:
Transcriptional Repression by Histone Deacetylases in Plants
If you peruse the current state of the art on these subjects, you will notice that following the general trend, Chinese authors are well represented. They lack some of the scientific (really unscientific) biases so entrenched here in the west. They are interested in results.
Here are a couple more papers that illustrate the genome+transcriptome+epigenome etc multivariate "wider canvas" new paradigm that are germane to the OP.
From Kew, not a research paper, but a note on one of their projects:
Plant Dormancy Transcriptomes and Epigenetic Control
Research paper:
Arabidopsis Paired Amphipathic Helix Proteins SNL1 and SNL2 Redundantly Regulate Primary Seed Dormancy via Abscisic Acid–Ethylene Antagonism Mediated by Histone Deacetylation
Research paper with a broader view of the same mechanisms in plants in general:
Transcriptional Repression by Histone Deacetylases in Plants
If you peruse the current state of the art on these subjects, you will notice that following the general trend, Chinese authors are well represented. They lack some of the scientific (really unscientific) biases so entrenched here in the west. They are interested in results.
S
sirius haze
hello, i was joking but yes the cotyledons seems strong, dark green and there is some violet/purple on the hypocotyl. you must have some incredible seedstock to play with, are you still collecting some landraces in India ? im planning to visit eastern states of India/ Burma border.
SKUNK # 1 seedlings
This is great thread, thanks Sam for the information. As Chimera already stated, it will be incredible when we are all finally able to share all the knowledge of this plant with each other, while not worrying about the limits of laws! Thanks to both you guys(and everyone else!) for keeping ICMag amongst the best sources of Cannabis information that are available today.
Sam I hope you don't mind me getting slightly of topic. It is a question pertaining to seed germination rates, so it is relevant. I asked this question here before but there was little interest.
I want to ask the question, do you think keeping all the weak seeds that barely germinate, and babying them with all sorts of unnatural germination methods, will cause more weak seeds in future generations?
I believe selection starts as soon as the seeds hit dirt. When I start seeds I wait no longer than 10-14 days after sowing the seeds and move on. I know there may be more seeds that pop up late, but why keep weak or slow (unless maybe they are rare seed stock of limited quantity?). I germinate directly into a soil or soiless mix, and give my seeds no special treatment. Straight from the seed fridge to the dirt.
I always figured healthy, quick germinating seeds would pass on genes for healthy, quick germinating seeds, and weak, slow seeds likely pass on genes for weak, slow seeds. If we want healthy seeds why do so many people go to such great(and unnatural) lengths to nurse seeds to life that otherwise would likely have been eliminated from the gene pool by natural selection?
One thing I think we all can agree upon, is that we would always want the most viable seeds possible. I would imagine after many generations of this type of selection we could have a very strong improvement in germination rates of Cannabis seed lines. I would love to have some input on the topic if anyone is interested.
Sam I hope you don't mind me getting slightly of topic. It is a question pertaining to seed germination rates, so it is relevant. I asked this question here before but there was little interest.
I want to ask the question, do you think keeping all the weak seeds that barely germinate, and babying them with all sorts of unnatural germination methods, will cause more weak seeds in future generations?
I believe selection starts as soon as the seeds hit dirt. When I start seeds I wait no longer than 10-14 days after sowing the seeds and move on. I know there may be more seeds that pop up late, but why keep weak or slow (unless maybe they are rare seed stock of limited quantity?). I germinate directly into a soil or soiless mix, and give my seeds no special treatment. Straight from the seed fridge to the dirt.
I always figured healthy, quick germinating seeds would pass on genes for healthy, quick germinating seeds, and weak, slow seeds likely pass on genes for weak, slow seeds. If we want healthy seeds why do so many people go to such great(and unnatural) lengths to nurse seeds to life that otherwise would likely have been eliminated from the gene pool by natural selection?
One thing I think we all can agree upon, is that we would always want the most viable seeds possible. I would imagine after many generations of this type of selection we could have a very strong improvement in germination rates of Cannabis seed lines. I would love to have some input on the topic if anyone is interested.
It seems (according to The Real Seed Company) that for example certain Himalayan varieties have seeds with different germinating times (meaning one variety has 'early' and 'late' seeds). This is also known from other plants and simply a survival mechanism; imagine there's late snow freezing all seedlings to death... the ones germinating weeks later prevent extinction. Other species even have seeds which will germinate a year or two later for about the same reason (not just late snow though).
Also, a good part of a seeds viability, sprouting behaviour, and vigour in the first days after germination initiation is in part due to the mothers tissue; but the germling is only ~50% mother, the rest is from a father. Don't forget, sometimes two weak parents can still give rise to a very vigorous offspring (p.ex. hybrid vigour as seen in extremis with hybrids from doubled haploid parents).
Also, a good part of a seeds viability, sprouting behaviour, and vigour in the first days after germination initiation is in part due to the mothers tissue; but the germling is only ~50% mother, the rest is from a father. Don't forget, sometimes two weak parents can still give rise to a very vigorous offspring (p.ex. hybrid vigour as seen in extremis with hybrids from doubled haploid parents).
That is touching on what I was saying about cytological inheritance earlier. I think most people, even well educated folks, don't realize the we humans, cannabis plants, most organisms, are little pieces of their mother 'infected' with their father's nuclear DNA. Your cells are literally your mother's cells, all the membranes, organelles, all the machinery of gene expression, this is all your mom's. Some of these things can then be modified by the new nuclear DNA, but in reality, you are a piece of specialized tissue from your mom's body, that has been modified by your dad's nuclear DNA.
Hi SeedsOfFreedom
Hey that is a great question that helps illustrate the points about the forces that determine form and behavior/function of an organism.
It has been beaten into our heads that "everything is genetic". If you observe a trait, you think "I wonder what gene or genes control that trait?". In the 20th Century, the practical success of this mindset was phenomenal, truly amazing in all fields of life sciences, from agriculture and plant breeding to human medicine.
In recent years though, it has become apparent that there is more to it than that. It turns out that the regulation of these genes is just as important as their presence, and that there are non-DNA, non-gene elements that encode information as important as that in the DNA.
Even the arrangement of the DNA can encode information. A crude analogy would be an old-fashioned dead-tree library. The information in the books would be the info encoded by the genes/DNA. You could though, especially in a big library, encode information by taking the individual volumes and arranging them into the shapes of letters, used then to make words, words to sentences, to paragraphs, ending up with a book made up of books.
Like previous paradigm shifts (like classical physics to modern relativistic/quantum), it starts with a very successful structure, that with closer and closer scrutiny, has some cracks. It is the exploration of these cracks (for relativity it was the failure to detect motion through the "ether" and the observed motion of the planet Mercury, for quantum it was blackbody radiation, discrete atomic spectra and radioactive decay) that leads to the new mindset that "fills the cracks".
The phenomenally successful gene centered mindset really started showing its cracks with the realization of the various genomic studies that have been on the forefront of biologic research in the past 30 years or so. If you remember, the Human Genome Project was trumpeted as a huge turning point in medicine, and would bring about cures for many, even most, human diseases. Although successful in a number of ways, compared to the expectations it is a dismal failure in regards to curing disease. Don't get me wrong, it was an important, essential step, but not the whole answer. Investigating this failure has led to a new understanding of the dance between genes, transcriptional machinery, epigenetic mechanisms etc. that determine form and behavior, and the manifestation of pathology.
Your "breeding for seed quality" illustrates this. Seed traits are controlled, in part, by information encoded in DNA, in genes. I think though, that a lot of the trait observed in a germinating seeds, (germ time, seedling vigor, maybe even sexual expression to some degree) are determined by non-genetic information in the epigenome and transcriptome of the seed. This information would be informed by the environmental conditions under which the seeds were formed, and the environmental conditions of the seeds after dehiscence until germination. I believe there are many factors involved in this, and would love to do research on this topic.
That wasn't what I meant but you're quite right too I guess.
What I meant were integuments, pericarp etc.
.
Thanks for the response guys, this is the type of scenario I speak of. This plant may have been great but took that long to germinate, if the breeders picked for quick germination, they may well have eliminated the seeds problems while retaining the vigorous plants and great smoke aspects.
The main problem I see with this proposed situation is the seeds germination controlling genes could be linked to others, or the exact same genes that actually control some of the the vigor and quality.
As for the epigenetic factors involved, this is part of the reason I usually use the soil or soiless mediums to start seeds, then hopefully any epigenetic mechanisms would recognize the soil/soiless environment as close to the earth there ancestors where in, thus triggering the same epigenetic response.
I know epigenetics are at play all over the place, but I think if breeders always keep germination environments close to the same, then eventually the seeds will at least breed true for strong fast germination in that environment. Whether epigentics will change germination rates is a new environment would be another great experiment as well.
Hi SeedsOfFreedom
There are genes that have major influence on seed characteristics, and some of them would be the same ones for "general vigor" of the mature plant.
From personal experience though, selecting from the quickest germinating, most vigorous seedlings does not achieve the goals you are looking for. Have you ever tried this?
Most long-time growers will tell you that if you plant 100 seeds, and cull all but the first 25 to germinate, you will end up with a bunch of males. Or, if you germinate 100 seeds, and select the 25 largest, most robust individuals, you will have 25 males.
As an experienced grower learns to use this knowledge and it becomes pretty easy to tell which seedlings are likely male and which are female even before they show any reproductive parts. With my lines, that I am very familiar with, I can tell with close to 100% accuracy which are which by the third week. So, if you take this into account, and select the quickest germinating females, their offspring do not (in my experience) have a different distribution of germinating times. This strategy also fails to improve general vigor (again, in my exp).
This pertains to variation within the variety. Different varieties, on the other hand, do have different germ rates, mostly due to pericarp characteristics like OO mentioned. I think you for sure could breed for this easily and effectively. I have no need to do this though, I germ in soil, and the tannins, microorganisms, enzymes etc in living soil provide an environment that makes even the thickest-hulled, stubborn seeds pop right open. I feel, but cannot say for sure, that thick-hulled seeds store better anyway.
Don't let me discourage you from experimentation, though. I hope you experiment around on your own and discover some useful information!
I have a bunch of ideas to explore along this line of thought, and intend to share them in a new thread sometime. I hope you will join the discussion when I do.
Thanks
There are genes that have major influence on seed characteristics, and some of them would be the same ones for "general vigor" of the mature plant.
From personal experience though, selecting from the quickest germinating, most vigorous seedlings does not achieve the goals you are looking for. Have you ever tried this?
Most long-time growers will tell you that if you plant 100 seeds, and cull all but the first 25 to germinate, you will end up with a bunch of males. Or, if you germinate 100 seeds, and select the 25 largest, most robust individuals, you will have 25 males.
As an experienced grower learns to use this knowledge and it becomes pretty easy to tell which seedlings are likely male and which are female even before they show any reproductive parts. With my lines, that I am very familiar with, I can tell with close to 100% accuracy which are which by the third week. So, if you take this into account, and select the quickest germinating females, their offspring do not (in my experience) have a different distribution of germinating times. This strategy also fails to improve general vigor (again, in my exp).
This pertains to variation within the variety. Different varieties, on the other hand, do have different germ rates, mostly due to pericarp characteristics like OO mentioned. I think you for sure could breed for this easily and effectively. I have no need to do this though, I germ in soil, and the tannins, microorganisms, enzymes etc in living soil provide an environment that makes even the thickest-hulled, stubborn seeds pop right open. I feel, but cannot say for sure, that thick-hulled seeds store better anyway.
Don't let me discourage you from experimentation, though. I hope you experiment around on your own and discover some useful information!
I have a bunch of ideas to explore along this line of thought, and intend to share them in a new thread sometime. I hope you will join the discussion when I do.
Thanks
This is one of the selection criteria for my breeding program, but all of my crosses are only 1 generation in so far, so it is hard to see any difference in overall germination rates. I would imagine by 3-4 generations in I may see a difference, or I will realize there is no connection between germination rates in parent plants, and germination rates amongst their offspring.
S
sourpuss
sourpuss and the pot factory
sourpuss and the pot factory
Heres what im thinking, 6 golden tickets! Put into random packs of your seeds. Better yet, you gm 6 seeds to grow into a golden ticket of course 1 is mine for coming up with the idea
We tour the factory and you chose the winner to takeover, along with your minions, midgets, whatever you use at your factory.
Really we just come sit in a room and smoke your dry sift and fly around
sourpuss and the pot factory
Heres what im thinking, 6 golden tickets! Put into random packs of your seeds. Better yet, you gm 6 seeds to grow into a golden ticket
of course 1 is mine for coming up with the ideaWe tour the factory and you chose the winner to takeover, along with your minions, midgets, whatever you use at your factory.
Really we just come sit in a room and smoke your dry sift and fly around
