tissue culturing Sarracenias

Mellard, David (dam7@ATSDHA1.EM.CDC.GOV)
Fri, 16 Aug 96 03:57:00 EST

On August 15, Peter Cole wrote:

< A question for any of the list's resident TC gurus - I've tried and
< tried, but I can't seem to get Sarracenia leaf tissue or rhizome borings
< sterile using bleach alone, even with concentrations up to 20% and times
< up to 30 minutes. Anyone know a good protocol? Should I pretreat the
< plants in some way perhaps? I've tried dosing them with systemic fungicide
< (Benlate,) 2 or 3 days before meristemming, but this doesn't seem to help.
< I'm using 1/6 MS and some plus 1mg/L IBA, but that's no help if I can't get
< the inocula sterile.

Hi Peter,

I'm no expert by any means but you might try adding a surfactant to the
chlorox/water solution. The surfactant will reduce the water tension, which
will allow the chlorox/water solution to penetrate the cracks and crevices on
the leaf's surface where microorganisms can hide. Pretty much any liquid
soap (a few drops) will do or you can use Tween 80 from a scientific supply
company. You also might try following the chlorox/water treatment with a
couple of rinses in 70% ethanol. A 70% solution (7 parts 100% ethanol, 3
parts water) is best since having water present increases the efficiency of
killing the microorganisms. If ethanol is available, you can use isopropyl
alcohol (rubbing alcohol) from the drug store, although it may not be as
effective as ethanol.

Thanks for the tip on your web site and please let me know whether or not
this fixes your problem.

David