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Thread: N. villosa/ N. macrophylla TC help

  1. #9
    pokie22's Avatar
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    fall under the aegis of proprietary knowledge...I am going to have to pick your brain then BB

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    BigBella's Avatar
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    Quote Originally Posted by pokie22 View Post
    fall under the aegis of proprietary knowledge...I am going to have to pick your brain then BB
    Pick away, Pokie . . .
    ôSý perchÚ l'autoritÓ dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    SimonTheSkink fiercedeity's Avatar
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    Quote Originally Posted by BigBella View Post
    Consider for a moment the expense of setting up a tissue culture laboratory, above that of, say, the average hobbyist with a cut-up Rubbermaid« tub; a collection of baby food bottles; a spray-bottle of bleach; and a Sacred Heart of Jesus votive candle. Consider then the expense involved in primary research and the myriad and often costly failures; and the very-western concept of intellectual property, which is seen as somehow legitimate in every other realm of human endeavor.

    As an aside, Nepenthes has been tissue cultured for decades; yet poaching is, arguably, as rampant as it always has been; and black markets still exist throughout Asia.

    Consider, also, how small the market is for carnivorous plants, which is, admittedly, far greater than when I began; but is still insignificant, when compared to orchid cultivation. Growers would naturally wish to maximize their profit in so small a venue.

    That said, I had moderate success with isolating the growing tip of Nepenthes hamata a few years back, for a Japanese client; and that involved the preparation of a carefully-cleaned 3 cm or so section of plant. At the time, I had ready access to fungicides no longer legal within the US, and to perhaps a dozen or more antibiotics and antimycotics at my disposal. It took serial re-plating to obtain a "clean culture;" but it can be done. The standard media was 1:3 MS with 8.8 ÁM 6-BAP; but meristematic tissue is a real coin toss, since you cannot be sure which growth phase the tissue is in when you make the attempt to establish it. Nepenthes also has a reputation for being "woody;" and there are some other medias, namely WPM, where growers have had some success.

    There are many papers involving in vitro culture of Nepenthes online. A twenty minute search should find a few. Still others are on costly academic databases; but abstracts are generally available. Check also the ICPS database for TC cultivation and the www.flytrapcare.com tissue culture forum for sources. There have been some threads within the last year or two . . .

    Thank you, I will look into those forums and see what I can find. I appreciate the help quite a bit.

  4. #12
    SimonTheSkink fiercedeity's Avatar
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    Quote Originally Posted by BigBella View Post
    1:3 MS with 8.8 ÁM 6-BAP
    Forgive me for my ignorance but you said to use 8.8 ÁM 6-BAP, ÁM being a measurement in micrometers which is length and not volume, how exactly does that work? Sorry for the onslaught of what must seem like foolish questions.

  5. #13
    i dont do pots. amphirion's Avatar
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    M= molarity, iirc.
    m= meters

    so 8.8 micromolar solution of 6-BAP
    Last edited by amphirion; 02-27-2013 at 12:48 PM.
    " You keep using that word. I do not think it means what you think it means." -Inigo Montoya
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    BigBella's Avatar
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    Yes; or, alternatively, 2ml/L of a standard 1 mg/mL solution of 6-BAP . . .
    ôSý perchÚ l'autoritÓ dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    BigBella, is it species specific in the amount of BAP you use? Thanks.

  8. #16
    BigBella's Avatar
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    Quote Originally Posted by DonH View Post
    BigBella, is it species specific in the amount of BAP you use? Thanks.
    Yes, in terms of all cytokinins, and not just limited to 6-BAP. Some genera, including a few Nepenthes respond to 0.1 ml/L solutions (0.44 ÁM) of 6-BAP, while others may require upwards of 2.0 ml/L or more (8.8 ÁM) for any effectiveness -- twenty times that lower rate. Aside from their role in cell division and the production of offshoots, they also prevent senescence in cultures and act as a "preservative" of sorts; and I routinely use it at low levels for the germination of seed. Others, including Heliamphora, seem to respond more effectively to TDZ (thidiazuron), once used as a chemical defoliant in the harvesting of cotton plants; and thought, for a time to be an anti-cytokinin -- but quite the opposite . . .
    ôSý perchÚ l'autoritÓ dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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