Tissue Culture / Micropropagation

Green SupremeGreen Supreme Plant ManagerHeaven BCPosts: 17,347
edited May 2012 in The Cutting Edge
Some info to insite interest. Peace GS

http://planttc.com/
Post edited by Green Supreme on
Nobody wants to plant the corn,everybody wants to raid the barn.
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Comments

  • Monseigneur StroganoffMonseigneur Stroganoff Posts: 4,378
    edited January 2008
    thats a cool link GS thank you




    ....wonder if it works on cannabis tho:chin:, heard its not that easy...

    gotto find that out
  • c-rayc-ray germinating Posts: 14,732
    edited January 2009
    we can do it!

    Micropropagation Part I Intro


    Micropropagation Part 1 - Intro cont.


    Micropropagation Part II - Getting Started


    Micropropagation Part III - Getting Started


    Micropropagation Part IV - Getting Started


    Micropropagation Part V - Making Media


    Micropropagation Part VI - Making Media


    Micropropagation Part VII - Making Media


    Micropropagation Part VIII - Disinfecting


    Micropropagation Part X - Disinfecting


    Preparing a Saucer Magnolia Explant


    there is a couple vids missing but he is apparently going to add them and some new ones soon
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • DOZEEDOZEE Herb-Smith Posts: 1,609
    edited January 2009
    fracken cool
    **DISCLAIMER: I am not currently, nor have I ever grown, smoked, or even seen real marijuana. All of the pictures posted here by me are not my own and I would never think of breaking any law of the United States, no matter how antiquated or stupid.**
  • c-rayc-ray germinating Posts: 14,732
    edited February 2009
    bananas



    aquatic plants



    a lecture in 3 parts







    supplies:
    http://www.phytotechlab.com
    http://www.caissonlabs.com
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • DOZEEDOZEE Herb-Smith Posts: 1,609
    edited February 2009
    Thanks for the Insperauto C-ray. Building slowly. hope to have a nice hobbie area.
    **DISCLAIMER: I am not currently, nor have I ever grown, smoked, or even seen real marijuana. All of the pictures posted here by me are not my own and I would never think of breaking any law of the United States, no matter how antiquated or stupid.**
  • Martian Junior Member Posts: 7
    edited February 2009
    Hi All.
    Splendid thread, been meaning to check this out, if one can make a suitable "agar" or sterile nutrient medium, we already have most of the other things needed.
    What a shit hot way of keeping a "strainbase" of plants your not growing at the moment, but may want to in the future.
    And what a challenge, I've not yet found anyone who has tissue cultured weed.
    Push that envelope!!!!!!!!!:nod:
    Ta Ta.
    Martian
  • c-rayc-ray germinating Posts: 14,732
    edited February 2009
    I am finding this all rather fascinating
    for instance yesterday I learned that sometimes it is benefiial to tissue culture old seeds rather than directly germinating them
    the process would go something like this:
    put some seeds in a jar with some 5% bleach water and shake for 5 minutes then carefully remove the shells from the seeds and put the embryos into a new jar of bleach water and shake again another 5 minutes then take out the now clean embryos with forceps in front of a laminar flow hood or in a glove box and drop into a jar with some sterilized nutrified agar with a dilute mix of nutrients, vitamins and hormones in appropriate proportions
    in the lab they use M&S salts + viatins + hormones but I see no reason why we can use what we are already using to grow cannabis but at a lower ppm like Lucas Formula + Growth Plus + Alg-A-Mic
    Growth Plus might be good for shoot production and BioBizz Alg-A-Mic for root production
    other things like urine and coconut milk might be useful too
    I've read the suggested pH is usually around 5.5
    and the agar concentration from 5-8%
    some cultures are started in the dark for a few weeks while some are started under lower light
    for the jars I think we can get away with tyvek + polypropylene in the lids for air exchange
    some humidity is probably good like maybe 70%
    I have read of successful cannabis tissue culture using leaf petioles, meristems (shoot tips) and nodes
    a node is basically a piece of stem with a meristem
    I have read that it is good to have a decent amount of tissue for good vigourous growth

    some reasons we need to figure this out
    one can hold 100's of cuttings in callus form in a small box and keep it in a non-growing buddies fridge for long term storage
    little bits of callus can be transported to wherever they need to go inconspicuously
    plants can be cleaned of pests and disease
    and many plants can be produced more rapidly

    post your results and experiences below
    vvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvvv
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • c-rayc-ray germinating Posts: 14,732
    edited February 2009
    [table]KH2PO4 ([URL="http://en.wikipedia.org/wiki/Potassium_dihydrogen_phosphate"]Monopotassium phosphate)|170 mg/l
    MgSO4 * 7 H2O ([URL="http://en.wikipedia.org/wiki/MgSO4"]Magnesium sulfate)|370 mg/l
    NH4NO3 ([URL="http://en.wikipedia.org/wiki/NH4NO3"]Ammonium Nitrate)|1650 mg/l
    KNO3 ([URL="http://en.wikipedia.org/wiki/Potassium_Nitrate"]Potassium Nitrate)|1900 mg/l
    CaCl2 * 6 H2O (Calcium Chloride Hexahydrate)|440 mg/l
    H3BO3 ([URL="http://en.wikipedia.org/wiki/Boric_Acid"]Boric Acid)|6.2 mg/l
    MnSO4 * 4 H2O (Manganese Sulfate Tetrahydrate|22.3 mg/l
    ZnSO4 * 7 H2O ([URL="http://en.wikipedia.org/wiki/Zinc_Sulfate"]Zinc Sulfate Septahydrate)|8.6 mg/l
    KI ([URL="http://en.wikipedia.org/wiki/Potassium_iodide"]Potassium Iodide)|0.83 mg/l
    Na2MoO4 * 6 H2O (Sodium Molybdate Hexahydrate)|0.25 mg/l
    CuSO4 * 5 H2O ([URL="http://en.wikipedia.org/wiki/Copper(II)_sulfate_-_pentahydrate"]Copper Sulfate Pentahydrate)|0.025 mg/l
    CoCl2 * 6 H2O ([URL="http://en.wikipedia.org/wiki/Cobalt_chloride_hexahydrate"]Cobalt Chloride Hexahydrate)|0.025 mg/l
    FeSO4 * 7 H2O ([URL="http://en.wikipedia.org/wiki/Iron(II)_sulfate"]Iron Sulfate Septahydrate)|27.8 mg/l
    Disodium EDTA|37.3 mg/l
    Agar|10 g/l
    Sucrose|30 g/l
    [URL="http://en.wikipedia.org/wiki/Glycine"]Glycine|2.0 mg/l
    [URL="http://en.wikipedia.org/wiki/Indoleacetic_acid"]Indoleacetic acid|1 to 30 mg/l
    [URL="http://en.wikipedia.org/wiki/Kinetin"]Kinetin|0.04 to 10 mg/l
    [URL="http://en.wikipedia.org/wiki/Myo-inositol"]Myo-inositol|0.5 mg/l
    [URL="http://en.wikipedia.org/wiki/Pyridoxine_HCl"]Pyridoxine HCl|0.5 mg/l
    [URL="http://en.wikipedia.org/wiki/Thiamin"]Thiamine HCl|0.1 mg/l[/table]
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • guest Posts: 24,389
    edited February 2009
    Nice thread!

    I've been wanting to spend time looking into this for a while. Yup C-ray, I've read about germing old or contam prone seeds (not just cannabis) In vitro (tissue culture).
  • dextr0 Junior Member Posts: 12
    edited November 2009
    ...stole this...thought it might b of some help to someone....
    Plant Tissue Culture for the Home

    Although technical procedures for aseptic culture of plant cells, tissues, and organs are as diverse as the plant material on which they are practiced, a simplified general procedure can be followed in the home. All that is needed are a few basic supplies which can easily be obtained. The procedures outlined in this section can be used in the home to propagate various species of plants, both easy (African violets, coleus, chrysanthemums) and difficult (orchids, ferns, weeping figs) to propagate.
    Medium Preparation
    For 2 pints of tissue culture medium, mix the following ingredients in a 1-quart home canning jar:
    1/8 cup sugar
    1 teaspoon all-purpose, soluble fertilizer mixture. Check the label to make sure it has all of the major and minor elements, especially ammonium nitrate. If the latter is lacking, add 1/3 tsp. of a 35-0-0 soluble fertilizer

    1 tablet (100 mg) of inositol (myo-inositol) which can be obtained at most health food stores 1/4 of a pulverized vitamin tablet which has 1 to 2 mg of thiamine

    4 Tablespoons coconut milk (cytokinin source) drained from a fresh coconut. The remainder can be frozen and used later.

    3 to 4 grains (1/400 teaspoon) of a commercial rooting compound which has 0.1 active ingredient IBA


    Fill the jar with distilled or deionized water. If purified water is not available, water that has been boiled for several minutes can be substituted. Shake the mixture and make sure all materials have dissolved.
    Baby food jars with lids, or other heat-resistant glass receptacles with lids can be used as individual culture jars. They should be half filled with cotton or paper to support the plant material. The medium should be poured into each culture bottle to the point where the support material is just above the solution.
    When all bottles contain the medium and have the lids loosely screwed on, they are ready to be sterilized. This can be done by placing them in a pressure cooker and sterilizing them under pressure for 30 minutes or placing them in an oven at 320oF for 4 hours. After removing them from the sterilizer, place them in a clean area and allow the medium to cool. If the bottles will not be used for several days, wrap groups of culture bottles in foil before sterilizing and then sterilize the whole package. Then the bottles can be removed and cooled without removing the foil cover. Sterilized water, tweezers, and razor blades, which will be needed later, can be prepared in the same manner.
    Plant Disinfection and Culture
    Once the growing medium is sterilized and cooled, plant material can be prepared for culture. Because plants usually harbor bacterial and fungal spores, they must be cleaned (disinfected) before placement on the sterile medium. Otherwise, bacteria and fungi may grow faster than the plants and dominate the culture.



    Various plant parts can be cultured, but small, actively growing portions usually result in the most vigorous plantlets. For example, ferns are most readily propagated by using only 1/2 inch of the tip of a rhizome. For other species, 1/2 to 1 inch of the shoot tip is sufficient. Remove leaves attached to the tip and discard. Place the plant part into a solution of 1 part commercial bleach to 9 parts water for 8 to 10 minutes. Submerge all plant tissue in the bleach solution. After this time period, rinse off excess bleach by dropping the plant part into sterile water. Remember, once the plant material has been in the bleach, it has been disinfected and should only be touched with sterile tweezers.
    After the plant material has been rinsed, remove any bleach-damaged tissue with a sterile razor blade. Then remove the cap of a culture bottle containing sterile medium, place the plant part onto the support material in the bottle making sure that it is not completely submerged in the medium, and recap quickly.
    Transferring should be done as quickly as possible in a clean environment. Therefore, scrub hands and counter tops with soap and water just before beginning to disinfest plant material. Rubbing alcohol or a dilute bleach solution can be used to wipe down the working surface.
    After all plants have been cultured, place them in a warm, well-lit (no direct sunlight) environment to encourage growth. If contamination of the medium has occurred, it should be obvious within 3 to 4 days. Remove and wash contaminated culture bottles as quickly as possible to prevent the spread to uncontaminated cultures.
    When plantlets have grown to sufficient size, transplant them into soil. Handle as gently as possible because the plants are leaving a warm, humid environment for a cool, dry one. After transplanting, water the plants thoroughly and place them in a clear plastic bag for several days. Gradually remove the bag to acclimate the plants to their new environment; start with one hour per day and gradually increase time out of the bag over a two-week period until the plants are strong enough to dispense with the bag altogether.
  • ljb Member Posts: 56
    edited December 2009
    [URL="http://www.scribd.com/doc/14571756/The-Biotechnology-of-Cannabis-Sativa"]The Biotechnology of Cannabis Sativa

    by Sam Zwenger, April 2009

    He's got a section on tissue culture. This is not something I've worked with, but you all might find the information useful.
    When 5-15 pieces of stem have fallen onto the plate, the sterile tweezers can be used to manipulate and move the pieces of Cannabis stem. They should be placed equidistant from one another and gently pushed down to ensure complete contact with the media. The lid can then be placed back onto the Petri dish. Parafilm, a stretchy plastic film, is wrapped along the edge of the plate and its lid. This helps in retaining moisture and keeping the contents sterile.
    also this:

    Full text at the link, from the [URL="http://www.pakbs.org/pjbot/abstracts/41(2)/13.html"]Pakistan Journal of Biology, April 2009: A micropropagation system for cloning of hemp (Cannabis sativa L.) by shoot tip culture ~ Ren Wang, Li-Si He, Bing Xia, Jin-Feng Tong, Ning Li and Feng Peng
  • TheForSakenTheForSaken Senior Member Posts: 108
    edited December 2009
    I remember BS, back at the old CW talking about this, I wonder why he never continued with this project?
    He may be the man to talk to for his knowledge on this issue, I believe last I read he had a factory doing experimental production. Then again, this was a few years ago my memory could be fadding, but I doubt it...
    TFS
    I hit him to get his attention.
    I shot him to calm him down..
    I killed him to reason with him...
  • guest Posts: 24,389
    edited December 2009
    I ask about that through a friend of his recently. And yep he did succeed. And at one time talked about selling elite gene-stock so one doesn't need to grow out a few hundred plants to find and elite. But last I heard, after he came back to Canada he didn't peruse it further. Though I could be wrong. Someone might chime in who knows better than me. But I think that's accurate enough. I don't think steve developed his own method, though I could be wrong, but I bet he has a great custom agar mix :) !
  • guest Posts: 24,389
    edited December 2009
    ps. lbj,

    great post as usual, thanks for that study. I have been researching this more over the past month,. I was thinking one could grow out hundreds of plants in a small space, over time, to find great breeding parents. Just keep all potential strains in petri until ready to grow and test. Thus one can keep say 200 strains of one variety alive until all 200 are tested. Then cull all inferior ones. That's not possible right now unless one has huge resources...
  • c-rayc-ray germinating Posts: 14,732
    edited December 2009
    I grow out hundreds of seedlings at a time every grow, in a single room, without tissue culture
    it is not that hard just got to keep an eye out for hermies and sneaky males

    imho tissue culture might be a good idea for saving clones long-term in a fridge or germinating old seeds without having to sprout them, but only for those who have access to a flow hood, a clean room and lots of patience...it is not a fast process
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • guest Posts: 24,389
    edited December 2009
    Yea but do you get to see their full potential and test them for cannabinoid content and ratio? By growing out say 20-30, or less, at once in say a 3x3 or 4x4 table one can see the full phenotype expression, and not cull unnecessarily. Plus, that means nearly anyone with limited resources could make amazing new varieties with very large genepools. Few people can grow so many in a room at once and still choose the best expressions.

    Keeping careful notes of cannabinoid content and %, along with whatever other traits one seeks to work is good. After the first 20-30 are tested one keeps them as tissue culture and grows out the next 20-30. This way, with time, anyone can properly breed with large genepools. Even with your method it seems your mostly relying on phenotype, but that's only 50% genetic and 50% environment. So you if change the environ, ie. someone else grows it, they will get a different phenotype profile than what was breed for. I know it might not be a big morphology difference, but races are won by centimeters. By using genotype morphology like chemotype, and other base morphologies, one can make uniform varieties for certain traits, and for medicine knowing very closely what the % of cannabinoid to expect one can track the total cannabinoid per grow and equate that as phenotype to the chemotype to build a nice graph with OpenOfficeOrg for example. Record keeping is really important IMO. Thus by increasing the THC content (for example) one knows they are getting better at growing (just and example). But in the end, choosing on % of cannabinoid is the best bet, if that is the only trait one seeks to positivity select or, using negative section at the same time to remove certain traits as dominate.

    Trying to positively select for more than a couple of traits is very hard and IMO one is best to stick to 2 or 3, max.
  • guest Posts: 24,389
    edited December 2009
    Laminar flow hood can be build by nearly anyone for max, $200-300.00 in parts. There are tons of How-to's online.

    Clean room should be not needed, AFAIK (am I incorrect?), one could use laminar flow hood with big homemade glove box for under $100.00. Besides I'll post a great how-to for building one DIY clean room. I build one and I put pics in the fungi fun section of this site. I think I even gave directions and info about positive pressure being important, that and a antechamber, also with new tyvek painter suit, surgical gloves and mask directly after a shower. Surgical shoe covers are great, but don't wear 'dirty' shoes inside, a sticky mat can help a lot here. Most all contamination (microbes) resides withing the first two feet from floor, unless air movement raises them up, etc. Once one is in the tyvek suit, with gloves, mask and hood on, with foot covers, apply at least 71% (or 90% is even better) isopropenal (sp?) alcohol to paper towel and wipe down arms and gloves.

    I used to do this exact process at least twice a week up until a copule of weeks ago, reason is for mycological reasons. I work with grain to grain, plate work on agar in petri dishes, etc. I pour my own agar mix in front of the laminar flow hood while its warm in the kitchen, and do plate transfers for choosing fungal strains in clean room most of the time in front of the laminar flow hood (which also helps create the positive presser), along with another pseudo-flowhood setup without care to the wind velocity, I use it for positive pressure, same filter as a laminar flow hood...

    Ive seen pics of people doing tissue culture and growing out the plant (Salvia) and cacti (Peyote) in large mason jars. They poured the agar into jar, than do tissue transfer and close jar. There is tyvek as a filter, or a Whatman syringe filter (that is what I use) , the former covers a small drilled hole (<1/8-1/4") in the plastic bell lid, and the latter can fill a hole, both solution are for gas exchange while stopping all microbes. Worked quite well and was cheap.
  • guest Posts: 24,389
    edited December 2009
    Yup,

    See a thread in fungi fun called "Gojo Playhouse", thats my cleanroom, my "playhouse" :)
  • guest Posts: 24,389
    edited December 2009
    The thread is kinda hard to find, so I thought I'd just post a link for those interested. I am not claiming this clean room is % microbe free, but I have done G2G and agar work on a bench, not in front of the laminar flow hood. So I know this works, but I also use a laminar flow hood or glove box in the clean room in case it ain't so clean. To truly have clean room it costs thousands and will provide no greater clean cultures, but more peace of mind I guess because it doubtful to loose it's seal from the outside than my clean room.

    In that thread I was using a HEPA filter/fan setup from Walmart and it worked well if I worked directly in front. But it only worked for week or two. Then I had to switch to real laminar flow hood filters, etc.
    https://www.cannabis-world.org/cw/showthread.php?t=4865
  • c-rayc-ray germinating Posts: 14,732
    edited December 2009
    who said anything about unnecessary culling... it is important to grow plants at least twice or 3x to see their full expression anyways, clones are easy enough to keep around
    not sure I see your point on how tissue culture will make growing plants take any less space
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • guest Posts: 24,389
    edited December 2009
    By breeding with positive selection for chemotype (ex. high TCH % to other cannabinoids) and cannabinoid content, at the same time as negative selection for unwanted traits, one should only need to grow each strain once, thus speeding up the process vastly (years it would seem). If a breeder also wants to positively select for maturation time, or yield, that's fine AFAIK, but going over about 2-3 positive selection traits (esp. if they are all phenotype traits) in hope to stabilize said traits as dominate is hard to impossible due to becuase lots of positive section traits will creat too many variables, etc.

    I think good positive selection traits to breed for could be: % of THC to other cannabinoids (chemotype), percentage of terpenoids to cannabinoids, yield, aroma, etc. Currently I plan to positively select breed for % cannabinoids and total cannabinoids, ratio of cannabinoids to terpenoids and yield. Generally speaking, it seems with increased cannabinoids comes increased terps, flavinoids, etc. Thus I hope to be knocking down 3 birds with two stones by simply choosing a high ratio of THC, and a low ratio of cannabinoid(s) to terps where terps will be the much smaller % of course, etc.
  • guest Posts: 24,389
    edited December 2009
    Tissue culture uses less space becuase your only growing like 20-30 strains at any one given time. All other strains are in petri dishes which are very small, or small mason jars, etc. Once a batch of 20-30 strains are grown one makes a note of each of the strains' chemotype and exact cannabinoid profile %. The grower would keep the previously grown batch of strains as tissue culture so the next batch of 20-30 can be grown out. At the end of growing out all 200 strains one can check notes and simply trash all inferior strains in petri dishes, mason jar, etc, keeping the others to grow out as moms or dads.

    I don't have it all figured out yet but suggestion is sound.
  • c-rayc-ray germinating Posts: 14,732
    edited December 2009
    breeding is a subjective thing, it would be easy if thc was the only thing we are interested in, but it is not...so that is why it is important to grow the plants more than once, and ideally all side by side, and even in different gardens by different gardeners... and then tested (bioassayed) by as many folks as possible
    the biochemical ratios will different in every scenario, I guarantee it, they will even change in time as the plant is grown a few times from clone
    seeds are highly variable, there is really no need for everything to be homogenous only that it is good
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • guest Posts: 24,389
    edited December 2009
    c-ray;92626 said:
    breeding is a subjective thing, it would be easy if thc was the only thing we are interested in, but it is not...so that is why it is important to grow the plants more than once, and ideally all side by side, and even in different gardens by different gardeners... and then tested (bioassayed) by as many folks as possible
    the biochemical ratios will different in every scenario, I guarantee it, they will even change in time as the plant is grown a few times from clone
    seeds are highly variable, there is really no need for everything to be homogenous only that it is good

    As I mentioned, a breeder shouldn't really have a goal of stabilizing and fixing as dominate more than 2-3 different traits, the more traits one is trying to fix as dominate the harder, and more impossible it becomes. And if one is using only phenotype traits then the situation is much more difficult becuase phenotype depends upon 50% genes and 50% environ.

    A bioassay is an objective test, often quantitative, which for us means no less than semi-quantitative TLC is sufficient, not by two stoners smoking it. And as I mentioned, if one chooses to breed for genotype only traits like the proportion of THC to CBD, etc, one only has to grow them once. Cannabinoid ratios (to each other) are controlled by genes, so no matter what environ the plants are grown in (within rational reason) the ratios will be very similar, close to the same. However, one can breed to change the % of cannabinoids to each other. In general, its the concentrations that change with environs, not the ratios. That is why I will breed for ratios, and ratio of cannabinoids to terp, it's fixed genetically, not bound by phenotype. I will also breed for a phenotype trait, yield, but it's not the main goal, only genotypes traits are. Thus one (me) only needs to grow them once, becuase the traits one is fixing are not phenotypes traits, they are genotype traits. This is the whole basis of chemotype I, II, III, etc, classifications, they don't change by environ so it's a good method to discriminate between genetic chemotypes of the same or different variety or species of Cannabis spp..

    I prefer homogeneous, aka, stabilized hybrid, IBL or landrace than F2's, etc. If I'm breeding then sure variation is good, but if Im trying to fix certain traits, then no, wide variation is not good.
  • guest Posts: 24,389
    edited December 2009
    I thought I would cite the info about ratios of cannabinoids being genetic and not phenotype. I didn't want to get my list of references because it hard to find, and I'm being lazy on Xmas eve day, so I just thought I put this up in case anyone was curious:
    Why does GW cultivate its own cannabis plants?
    A.


    The absolute requirement for a plant-based medicine from a regulatory point of view is "control of starting materials". A drug in its manufacture goes through many processes, each of which need to be monitored and strict quality controls applied. This process control and QC would be invalidated if the starting materials were of poor or inconsistent quality. Laboratory analysis of GW's selected chemotype lines demonstrates that the cannabinoid ratios are very consistent. Such high levels of consistency are unusual in plants and are very important in applications made to medical regulatory authorities.

    GW's foremost consideration therefore is the cultivation of highly consistent plants with defined cannabinoid ratios. We have a number of chemotypes (varieties characterised by their chemical content) chosen for their composition and morphological traits.

    GW cultivates cannabis in order to be able to control the starting materials for the in-house production of GW's cannabinoid pharmaceutical products. GW does not distribute herbal cannabis to outside researchers or institutions.
  • guest Posts: 24,389
    edited December 2009
    This is just my un-scientifically based opinion, but nothing in the world will deter me from beleiving that two different siblings can have different effects. This has been tried tested and proven by experience.
    Peace and Merry x mass
  • guest Posts: 24,389
    edited December 2009
    Hey,

    I think I didn't explain it well, sorry. When you write "different effects" it seems to me you mean cannabinoid content, ie. more THC in sample A, and not a noticeable difference in cannabinoid ratios (to one another) if it's a IBL. The concentration of THC is phenotype (ie. 50% genetic and 50% environmental), the ratio of THC to CBN is genotype (all genetic).

    The ratios are not stable between sibling unless the variety itself is stabilized like IBL for example. But using cannabinoid ratio to each other and terpenoids of the same strain is genotype so it fluxes little between different environments. Sam Skunkman accomplishes that by backcrossing female tetraploid plant to the original mother diploid plant, and maybe one more inline cross, I have to check my notes. Regardless, the result being a very stable line (IBL) by cananbinoid ratios, which is important and good to know for the reasons I already mentioned (ie. charting total cannabinoid progess in same environ, etc, etc).





    HTH
  • c-rayc-ray germinating Posts: 14,732
    edited December 2009
    pretty sure gw pharma grows from clone, but you probably already know that being one of the 'top experts in Cannabis spp.'
    "One cannot develop taste from what is of average quality but only from the very best."
    Johann Wolfgang Von Goethe
  • guest Posts: 24,389
    edited December 2009
    Cray,

    They are breeding all the time (GW and HortaPharmBV). The seeds he will sell will be polyploidal (triploids :( ) As were the seeds he just this year imported to US for research at Cal U. with blessing of DEA! Check your PM in 5 minutes, I just wrote a long, very well referenced, and totally correct presentation about this very topic. Tho I don't want to share it in public yet.
  • guest Posts: 24,389
    edited December 2009
    Ill share this much publicly at this time:

    It's because he can only patent a plant if it's genetically unique, visa-vie, polyploid mutation and breeding to sterile and inferior genetically, ala. triploids! This is done so people like us can't make F2's or crosses with his feminized triploid seeds. Supposedly many Dutch breeders are planing on having Sam help them all create triploid seeds! So a grower would pay more for inferior genetics, and be forced to buy new seeds for each grow, unless one purchases clonal material, tho there are major rule on it's usage...or I guess people could re-veg the seed strain, but I've never liked the idea of re-vegging and flowering again.

    {EDIT} I made typos and wrote "diploid", when I meant to write "triploid", that is now fixed, sorry.
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