>
> Does anyone have any hints about tissue culturing Nepenthes or other CP's?
>I have some growing tips from Nepenthes burbidgea that I got off of at our
>local CP Society meeting and I'd like to try tissue culturing them in the
>next day or two. I have all the equipment and materials and some experience
>(I'm a high school Biology teacher) but I wonder if there's anything
>special I should know before I attempt this. >
The biggest problem I found with tissue culturing most ornamentals including
CP was phenolic oxidation. You should try to use a citric/ascorbic acid mix
(100mg/liter each) as a wash after you have sterilized the tissue to reduce
the browning and add 1 gram per liter of charcoal to prevent phenolic build
up in the media. Sterilization can be difficult, I would try 5% Chlorox (get
the good stuff, do not buy generic bleach) with 1 drop Ivory or similar soap
as a wetting agent.
Cut the pieces with 1 inch of excess tissue on the stems and clean off
leaves, bracts, etc. Place the pieces in a petri dish with the sterilizing
solution for 5 minutes. Using a forcepts transfer the pieces to a petri dish
with the ascorbate/citrate mix for five minutes then a dish with sterile
water before placing on the media.
The next biggest problem is sterile technique. If you are not familiar with
it practive handling pieces of other tissue to get the feel of the tools. Be
sure you dip the tools forceps, scalpes, etc. in 95% ethanol and flame them
between each use. I also autoclave the tools before I use them.
Good luck with the project. You might also want to try node segments with
dormant buds, they are less tender than shoot tips and with cytokinins in
the media can be stimulated to grow if properly handled.
Tom
/---------------------------------------------------\
| Tom Hillson | Iowa State University |
| Computer Coordinator | Experiment Station |
| hillson@iastate.edu | Room 17 Curtiss Hall |
| (515) 294-1543 | Ames, IA 50011 |
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