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Thread: Nepenthes seed in vitro

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    Jeremiah Harris's Avatar
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    Hello

    What is the best way to sterilize Nepenthes seeds. I tried dipping them in 70% alcohol then soaking them in 10% bleach solution for 10 minuets then rinsing them off in pure water but that seemed to have killed the seed embryo.

    What recipe of MS, PPM and BAP would work best for Nepenthes seed?

    Any help is greatly appreciated
    -Jeremiah-

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    Never done tissue culture before. Good luck man. [img]http://www.**********.com/iB_html/non-cgi/emoticons/new/smile.gif[/img]
    You have just recieved the Amish Computer Virus. Since the Amish don't have computers, it is based on the honor system. So please delete all the files from your computer. Thank you for your cooperation.

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    Try eliminating the alcohol. My guess is it is this step that killed the embryos. When I was working with orchid seed in vitro we never sterilized with anything but 10% chlorox soltion.

    Good luck.

    Phil

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    I don't think Jeremiah is tissue culturing, but raising the seed in flask, as an orchid breeder would do.
    I know one orchid flasker who claims that hydrogen peroxide is an effective replacement for the 10 percent clorox wash. How he dilutes the peroxide is unknown, but claims it is gentler on the seed.
    As for agar formulations-have you investigated some of the commercial pre-packaged formulas available for orchid growers...say something formulated for Cymbidiums, Cattleyas or Paphs.
    I have no idea what the nutrient requirements for Neps would be. I'll be watching this thread closely to find out. Tony?

    Trent

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    When people say 10% chlorox wash, it means that 1 part of chlorox has been diluted in 9 of water? or the final concentration is 10%.
    In our part of the world, we have this chorox equivalent called whiteking and in the bottle says it has 4.9 mls of pure sodium hypochlorite per 100 mls of water. In other words, the concentrated stuff is only 4.9%. If we dilute that 10 times, we'll get .49%.

    How much sodium hypochlorite is there in pure Chlorox??

    Thanks

    Gus

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    Very interesting indeed Jeremiah! I would like to try this myself. Keep us updated. Mabey some pics

    Gus,
    It's 6% here in the states.

    Robin

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    Me too, Robin. I would think there would be a higher germination rate, and more surviving seedlings in vitro as compared to sowing the seed on sphagnum and peat.
    Definitely something worth looking into.

    Trent

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    Trent, interesting hypothesis, but I think it's a case of six of one, half a dozen of the other. Both have their advantages and draw backs, and the end result after a year or two is often not that big.
    Demystifying Nepenthes: http://www.nepenthesforeveryone.com

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