Antw: Re: tissue culture!

Klaus Peper peper at med-ph.uni-sb.de
Fri Sep 13 08:28:16 EST 1996


Wiel Mattheij wrote:
> 
> > I have always used Tween 20, sorry if I didn't mention it originally.  I
> > have tried all the plant parts, and get the stems remaining green on
> > occasion and the leaves very occasionally, all other plants parts (
> > tendrils, pitchers, flower parts are killed in even the weekest
> > sterilants).  The 3.5% NaOCl may indeed be to strong for the plant, but
> > lowering the concentration won't help because even with an weekly spraying
> > with Benlate, a benlate dip, an ethanol dip and sterilization (with Tween
> > 20) the plants are still not coming up sterile.  The plant is insectivorous
> > and grows in damp conditions...  and nothing seems to kill the fungus (and
> > by the way it's not my bench technique before any one suggests it) - so what
> > I am really looking for is unusal and effective ways of sterilizing them,
> > for example has anyone had sucess with ultrsound?
> >
> > Ta Ta
> > Lindsey
> 
> I think it will be difficult to find an alternative sterilisation method for your plant material. The best way is to
> find a method where your plant material has some advantage above the (probably) internal infection of the
> fungus.
> You could try adding Benlate to your medium, if you didn't try it before. I don't know if Benlate can be
> heat-sterilised, if not, try adding it filter-sterilised.
> An other possibility is dropping the sugar content (even to 0%) of your medium in combination with high
> light illumination.
> You probably will need the fastest growing parts (shoot tips) of your plant and regular transplanting to
> fresh medium to accomplish this advantage. Maybe the application of gibberellins can be of use, to
> promote cell expansion.
> 
> Wiel Mattheij
> The Netherlands
> e-mail: wielmm at worldaccess.nl

First try to get a clean plant. If you grow them in pure shagnum or a
sterilized artificial medium in a terrarium, sealed with a plastic sheet, 
the new growth of the plant is pretty clean. Don't try to sterilize the 
complete plant. Take off new shoots or buds, sterilize them and try to 
get them in culture. If there is still contamination, try to peal off 
same scales from the bud. The proper technique depends on the plant.
Try dormant rizomes as in venus fly trap, or something else which has a 
meristem in it. Don't try to get down to the meristem, it is very 
difficult or almost impossible to find the right conditions for meristem 
culture. If you establish a method, please tell the internet people.

Good luck -- Klaus Peper
-- 
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