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Thread: Maybe crazy tissue culture experiment with Nepenthes seeds

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    BigBella's Avatar
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    Quote Originally Posted by Vidyut View Post
    Edit: Since you have done extensive TC, do you know if I can consider my cultures to be reasonably sterile and safe from contamination if nothing has shown up in 13 days? Also, should I worry about germination also resulting in contamination from inside the seed? Does that sort of a thing happen? (I am asking in the sense of whether I should be ready to sub culture at a moment's notice even now or I can breathe relatively easy...)
    You can be reasonably certain that the cultures are "clean" after almost two weeks. Contamination is usually obvious well before then.

    If the seed was intact, there should be no issue; though, you should have some fresh media in hand, in case you need to subculture any of the plants. Contamination, if any, can rapidly spread.

    Be patient as well. The fastest germination that I had ever experienced was just short of two weeks; and the longest, several months . . .
    Last edited by BigBella; 05-08-2018 at 10:28 AM. Reason: Typing with my feet . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

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    I have germination in the drosera seeds planted on sphagnum. Nothing in the TC. Neither contamination nor germination. No germination of nepenthes in sphagnum either.

    Anyone know what kind of a difference in germination time is normal between regular media and TC?
    Balcony farmer, carnivorous plants, DIY, sustainability, socio-political commentary India.

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    Quote Originally Posted by Vidyut View Post
    I have germination in the drosera seeds planted on sphagnum. Nothing in the TC. Neither contamination nor germination. No germination of nepenthes in sphagnum either.

    Anyone know what kind of a difference in germination time is normal between regular media and TC?
    In my experience, the in vitro seeds were the first to germinate, more than 75% of the time -- by an average of twelve days. Nepenthes seed generally have a short shelf-life, especially lowland species; but it also takes quite a bit longer than most Drosera to germinate . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Vidyut's Avatar
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    4 seeds of nepenthes maxima "giant wavy" germinating on sphagnum. Nothing in TC yet. No fungus, no germination. But some seeds seem.... swollen.

    Several other nepenthes seeds are also starting to germinate on sphagnum - very early hints of green.

    Nothing in TC
    Last edited by Vidyut; 05-13-2018 at 09:47 AM.
    Balcony farmer, carnivorous plants, DIY, sustainability, socio-political commentary India.

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    Quote Originally Posted by Vidyut View Post
    4 seeds of nepenthes maxima "giant wavy" germinating on sphagnum. Nothing in TC yet. No fungus, no germination. But some seeds seem.... swollen.

    Several other nepenthes seeds are also starting to germinate on sphagnum - very early hints of green.

    Nothing in TC
    Be patient with the seed in TC. Sometimes, it is helpful to keep the cultures in the dark for a period of 7-10 days after sowing. That has proved successful for me in the past; and has generally "sped" germination by about ten days, over those under light.

    Also, wasn't 50% MS used as germination media? That can be an issue; it's a bit strong. In the future, try 1:4 or 1:3 and soak the seed in distilled water overnight before sowing . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Vidyut's Avatar
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    Yes. Being patient as long as it takes to see germination unless there is contamination - in which case I am prepared to replate the seeds if I can. The delayed germination may also be because my media is not how they want it given that I've used too much MS.
    Balcony farmer, carnivorous plants, DIY, sustainability, socio-political commentary India.

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    Most of the seeds on sphagnum had at least one seed germinating slightly over 2 weeks. It is still one day short of 3 weeks.

    No action in vitro, but not worried yet. From all accounts it seems to be the sphagnum seeds that are on the early side.
    Balcony farmer, carnivorous plants, DIY, sustainability, socio-political commentary India.

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    I had a minor accident that led to one of the TC containers falling to the floor and cracking open. I took the seeds out and rinsed them and have them soaking in water to sterilize and put in TC again.

    Gave me the opportunity to look at the TC seeds and medium with a magnifying glass. Still no signs of contamination. I checked the pH, but my pH meter appears to be dead. According to strips, the pH is between 5 and 6 - which seems fine.

    Other containers as usual I think - no contamination, no germination, though I haven't looked at them too closely yet, while I was busy rescuing these seeds. This is now getting boring.
    Balcony farmer, carnivorous plants, DIY, sustainability, socio-political commentary India.

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