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Thread: Tissue Culture

  1. #9
    Drew's Avatar
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    i know nothing about this stuff so pardon my intrusion but the setup u have looks like a drug lab

    Ps.. i wach a lot of cops, csi, etc

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    Yes, Cristal, there is so much stuff you will not find in the literature and sometimes is vital for the success in TC. Michal does have most of his plants in vitro from seeds, but not all of them. I think about 30% comes from direct organogenesis. As far as I am concerned, I still try to find propagation protocols using other material than seeds. The reason is simple- some seeds are very hard to get. If you know how to propagate leaf/flower stalk/root explants you have unlimited amount of plant material at hand:-)
    You asked about the perfect time to deflask seedlings onto subcultre hormonal medium. Well, I put most of my TC seedlings or plants directly in regular substrate. The optimal time varies. For example, I got seeds from this mother plant of ascendens

    When she produced decent amount of fertile seeds in the summer, I thought of using TC instead of regular sowing to propagate this plant quickly (I love this species:-) ). They sprouted for me in 10 days,but some time later the seedling started dying due to overheating in my absence. When I returned I had to act promptly to save last 5 last seedlings (out of 40). So I had to deflask them prematurely. Out of 5 I think 4 will survive:


    With d. burmannii I could keep the seedlings maxinum of 2,5months in the jar, then it was too crowded and I had to deflask them, they look happy now:


    Drosera anglica I propagate from leaf explants now I know how to do it, so it is easy. Even WITHOUT hormones I got multiplication ratio of 14 new plants per leave. After 2 months in TC i deflasked the rootless plants on substrate where they rooted spontanneously:


    But of course, I always keep a back-up sterile material in subculture. You can do this at anytime, as long as the size of the plant allows it.
    For example, part of burmanni seedlings I put on hormonal medium into subculture. So far (20 days) I see no propagation via morphogenesis, the plants keep growing and flowering as in ordinary substrate:


    On the other hand, plantlets from leaf explants of d. anglica, put onto hormonal medium, started to form adventitious buds on leaves again. I subcultured 5 plants. With the thought in mind that hormones increase propagation ratio per leave, I should have this jar soon filled with plants:


    So the best way to make TC is to start from seeds. It is easier. When you have grown seedlings, you can use their sterile tissue for subcultures via other parts of the plant, with no neccessity of annoying sterilization! Since some seeds are hard for me to get (or sprout), I keep on trying other parts. I recently managed to initiate flower stalk of d.venusta and leaves of d.rotundifolia and d. collinsiae 'Faryland'. I keep on trying it is fun:-)

  3. #11
    Tropical Fish Enthusiast jimscott's Avatar
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    Carnivorous Blake has had some experience with TC. So has Jason Koch.

  4. #12
    What is and what should never be Crissytal's Avatar
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    Drew: The government probably thinks I do too ha. I had to order some lye recently to break down some hormones. I'm sure I've been flagged.

    klasac: Wow! Thank you for all of the detailed pictures and information! That's why I am so interested in TCing explants, I have almost an unlimited amount of tissue while seed is more difficult to come by. I just did some P. 'Titan' flower stalks. So far so good. It's only been a few days though. If I can only ever figure tissue out. Thanks for the deflasking information. I'd probably be better off letting the seedlings grow and get the tissue from them instead of moving the entire seedling. At least that's what I'm thinking at this point.

    Beautiful ascendens! I was lucky enough to get ahold of some seeds. I have a few popping up now (not invitro yet). Good luck with your remaining 4/5 seedlings. I hope they do well for you! The D. burmannii and D. anglica have done amazing. Good luck with your experiments! I will continue to experiment as well.

    Jim: Thanks!
    Where do we go when we just don't know,
    And how do we relight the flame when it's cold?
    Why do we dream when our thoughts mean nothing,
    And when will we learn to control?
    --Godsmack

  5. #13
    klasac's Avatar
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    Thanx, Crystal!
    Of course, you dont need to use the whole sterile seedling/grown plant. I only like to do it this way, because you can almost immediately see, how the actual whole plant reacts to the medium with hormones (flawless growth vs. slow growth or etiolated leaves, etc)...and, at the same time, the propagation is in progress :-) If the plant complains about the hormones you see it and next time use other mixture/ratio/hormone type.
    Thanx for the compliments on ascendens, it is an old big plant and produces seeds for me:-)
    I recently got my hands on ascendens seeds, the red form, which is the biggest of all. Cant wait to see the plants sprout.
    Good luck on your seedlings and plants! :-)

  6. #14
    klasac's Avatar
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    Hi Crystal! (and others, of course)! I got some new CP material sprouting:
    localised D. spatulata seedlings (no hormones):


    d.nidiformis seedlings (no hormones):


    d.collinsiae 'Faryland' (2 leaf explants on media with NAA and Kinetin) , swollen and producing minute adventitious buds:


    I am particularly happy for the former one, since leafs are hard to make work sometimes. Will report back with more such as d.tokaiensis, d. affinis and others:-)

  7. #15
    What is and what should never be Crissytal's Avatar
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    Hey klasac!

    Very nicely done!! So many germinations! Looks like everything is doing very well for you. This might be a silly question, what happens if seed are started on media with hormones? Every reference I have found simply says not to with no reason. Does it inhibit germination?

    Congrats on the explant success! I haven't figured it out yet. Lately I've been getting the material sterile, but also killing the explants. Back to the drawing board lol. What is your procedure on the D. collinsiae leaves if you don't mine me asking? Did you do anything ahead of time, like fungus treatment or anything before beginning the sterilizing procedure?

    Here are some of my updates (sorry for the picture quailty, taking pictures of seedlings inside jars isn't easy LOL):

    D. collinsae


    D. felix


    VFT seedling


    Drosophyllum


    Gemmae to see if it could be done:
    D. roseana


    D. pulchella x occidentalis


    I'm looking forward to your updates on D. tokaiensis, D. affinis and your others!
    Where do we go when we just don't know,
    And how do we relight the flame when it's cold?
    Why do we dream when our thoughts mean nothing,
    And when will we learn to control?
    --Godsmack

  8. #16
    CPlantaholic's Avatar
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    very cool to see both of your tc projects. I'm starting to get jealous...
    Visit The Sundew Grow Guides: http://www.growsundews.com
    New- Drosera video tours & other sundew info, now on YouTube!

    Happy Growing!

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