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Thread: Nepenthes in vitro . . .

  1. #61
    pokie22's Avatar
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    So, you like to snuggle under the blankets too.......

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    Quote Originally Posted by pokie22 View Post
    So, you like to snuggle under the blankets too.......
    That old photo was meant to suggest a New Year's Day hangover . . .
    “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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    zesty. BioZest's Avatar
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    I've seen this with Venus Fly Traps and sarrs, but not with neps, although I guess it's pretty much the same idea.... very interesting. Best of luck with the seedlings!

  4. #64
    Jimmy's Avatar
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    BigBella,

    An off-topic TC question... what would cause Nepenthes to turn brown and die part-way through germination in vitro? Assuming a standard media of 1/3 MS with vitamins and 25g/L sucrose with no PGRs.

    And while I'm asking, have you used activated charcoal in any of your cultures? I think I remember seeing in some literature that it helps in Nepenthes TC and it certainly has its place in orchid TC... BE also has a video that shows some of the Nepenthes TC stock and some cultures have the charcoal while others do not.

    Thanks!

    -Jimmy
    "I'm just a scientist without the proper documentation."

    My Growlist: http://www.terraforums.com/forums/sh...d.php?t=113724

  5. #65
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    Quote Originally Posted by Jimmy View Post
    BigBella,

    An off-topic TC question... what would cause Nepenthes to turn brown and die part-way through germination in vitro? Assuming a standard media of 1/3 MS with vitamins and 25g/L sucrose with no PGRs.

    And while I'm asking, have you used activated charcoal in any of your cultures? I think I remember seeing in some literature that it helps in Nepenthes TC and it certainly has its place in orchid TC... BE also has a video that shows some of the Nepenthes TC stock and some cultures have the charcoal while others do not.

    Thanks!

    -Jimmy
    Jimmy, the death of seedlings in vitro can occur for a few reasons -- chiefly among them, sustained higher temperatures (I rarely keep anything in culture above 20˚ C); though some seedlings simply spontaneously "abort" upon germination. That unfortunately does happen.

    In terms of the use of activated charcoal (AC), I have used it for a number of purposes. It removes plant exudates (such as phenols and excess PGRs) from media; to promote root formation (partially due to the darkness provided); and it diminishes unwanted callus growth. I used AC on a handful of occasions with Nepenthes seed (having read some of that same literature); though I was unimpressed by its supposed benefit to germination media. I had far better results, by nearly a third, by eliminating it. Its chief drawback is that it all-too efficiently removes organic nutrients and PGRs from media, especially auxins for some odd reason (one paper I had read suggested adding anywhere from 10-100x the normal amount if "high" levels of activated charcoal were present to offset that effect). AC also affects the given pH of media, by increasing it by about .75.

    If you insist upon using it, get some laboratory grade AC from a laboratory supply (Caisson Labs, for example); and don't bother to purchase some pecker-wood charcoal supplements from health food stores. You never know what you're getting; and the AC for tissue culture use has to be precisely prepared . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

  6. #66
    Jimmy's Avatar
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    Thanks for the detailed reply, BigBella! Temps may definitely have been an issue, as they were kept in about 80-degree-Fahrenheit temps after I set up the cultures... There were other factors too, but I wanted to ask if there are any key things to watch out for and you certainly helped!

    That's an interesting remark on AC... I'll keep it off my list and out of my cultures for a while until I have more room for experimentation. I'm still relatively new to TC and I'm more interested in the science of it and the degree of control it allows for plant growth, but I'd rather be TC'ing Nepenthes than anything just because they're my favorite. I may try orchids one of these days...
    "I'm just a scientist without the proper documentation."

    My Growlist: http://www.terraforums.com/forums/sh...d.php?t=113724

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    Quote Originally Posted by Jimmy View Post
    That's an interesting remark on AC... I'll keep it off my list and out of my cultures for a while until I have more room for experimentation. I'm still relatively new to TC and I'm more interested in the science of it and the degree of control it allows for plant growth, but I'd rather be TC'ing Nepenthes than anything just because they're my favorite. I may try orchids one of these days...
    I have no intention of denigrating activated charcoal's use in tissue culture; it's just that there is a mistaken, though popular notion among hobbyists, that it is simply an inert additive. Some AC even contains some trace amounts of PGRs, even after processing; and I have used it to encourage rooting in genera that are occasionally difficult -- some highland Nepenthes and Heliamphora, for example.

    Some Nepenthes species -- N. rajah, in particular, though for no apparent reason -- are long-lastingly affected by some PGRs (especially BAP); and activated charcoal plays a beneficial role in diminishing it from the media -- and even withdrawing some of it from the explants themselves . . .
    “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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    Here is a recent update on a second batch of 2012 seed, plated in December. All are on 1:3 MS with 3% sucrose and 1-2 mg/L 6-BAP . . .

    Nepenthes attenboroughii 5 February



    Nepenthes villosa (Tambuyukon)
    Last edited by BigBella; 02-06-2013 at 12:06 AM.
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

  9. #69
    Aspidistra Pilot mato's Avatar
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    In light of the forum's NASCAR fantasy league, "Git her done."
    Truth as Circe. Error has transformed animals into men; is truth perhaps capable of changing man back into an animal?

  10. #70
    Hear the Call of Nepenthes carnivoure12's Avatar
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    Lots of beauties coming along David! Can't wait to see how the progress.
    -Carnivoure12
    →Growlist

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