Antw: Re: tissue culture!

Wiel Mattheij wielmm at worldaccess.nl
Mon Aug 26 16:49:35 EST 1996


> 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







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