Re: Tissue Culture

From: Peter Cole (carnivor@flytrap.demon.co.uk)
Date: Tue Feb 17 1998 - 17:19:24 PST


Date: Wed, 18 Feb 1998 01:19:24 +0000
From: Peter Cole <carnivor@flytrap.demon.co.uk>
To: cp@opus.hpl.hp.com
Message-Id: <aabcdefg633$foo@default>
Subject: Re:  Tissue Culture


> Hello TC - experts !
> I have managed to establish some cp (D.regia, D.petiolaris, D.falconeri,
> Da.californica ...) from seeds in vitro. They are doing fine but so far I
> have failed
> to multiply them. I have tried cuttings but they always died after a while.
> If you have any advice or know some tricks please let me know.
> Thanks a lot for your help !
> Kind regards
> Jens

        I don't have any trouble multiplying D.regia if using whole leaves
        plucked from the in vitro plantlets and recultured - these will bud
        up at the base and form new plantlets which can be plucked apart,
        etc. Plucking whole leaves off seems more succesful than cutting
        them, though sometimes leaf tips or sections will work as well, so
        you could try cutting leaves in half. I think cutting them damages
        the tissue more and causes it to produce harmful phenols, as the
        medium around the cut surface often discolours and the cut edge
        turns black - the main techniques that can help reduce the damage
        are any of these:

                - keep flasks in dark for first 72 hours (to limit phenol
                        production)
                - add 2-5g/L activated charcoal to the medium (to absorb phenols -
                        you can get this from tropical fish shops)
                - reculture onto fresh medium after 72 hours (to replace
                        contaminated medium)

        It is certainly not a sp. that seems as amenable as many of the
        weedier S.African/Australian spp. to fine shredding onto the medium,
        but not difficult.

        Also new plantlets will often emerge spontaneously from the roots if
        you reculture whole plants in fresh medium after letting them
        deplete the original for 12 weeks+ (prompts rooting.) This seems
        irrespective of which medium they're in (I've tried MS, Knudsons,
        Vacin & Went.)

        I haven't had much luck with petiolaris - I don't like working with
        seeds (fiddly and I always seem to overdo the bleach), and on the
        few occasions I've managed to sterilise leaf tissue, it has browned
        and died in a few days. I guess it doesn't like the bleach :(
        Ooh - D.falconeri - I'm envious :)...

        Darlingtonia - I have to confess I've never had to sterilise (found
        a vesutor flasked plant in the local garden centre a couple of years
        ago, and worked from that :) It seems quite prolific once it gets
        going - the clumps can be pulled apart with tweezers, but I don't
        know how long it takes to get going from seed. It never seems to
        make bigger (adult) pitchers, but you can make many
        one-year-old-seedling-look-a-likes quite quckly. It needs to be
        starved in depleted medium for couple of months to get it to root I
        find, but you don't need many roots to transplant to soil safely.

        Hope this helps - happy cloning,

                        Peter.
+++ Peter Cole, 17 Wimmerfield Cr.,Killay,SWANSEA SA27BU,WALES,UK +++
mailto:carnivor@flytrap.demon.co.uk - http://www.flytrap.demon.co.uk/
++++ Carnivorous Plants, seeds and tissue culture kits for sale ++++



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