We are working with NaDCC for sarracenia sterilization. It is working great and should produce good results for Darlingtonia sterilization. Please see the sarracenia tissue culture section in the forum for up to date details. Darlingtonia tissue can be very hard to sterilize, the following techniques will produce favorable results but with a high failure rate. The NaDCC should work most of the time.
Darlingtonia californica Tissue Culture
Tissue Culture Difficulty: Seeds - Moderate: Leaves/Rhizome - Very Hard Usable Parts: Leaves, Rhizome, Seeds
Sterilization: Seeds:
15 min. bleach | 2 - 5 min. rinse or 2% PPM for 4 hours
Leaves w/rhizome:
Rinse in water | Alchohol dip | 2 min. peroxide | 35 min bleach | 3 - 5 min rinse| Soak in 5% PPM* for 2 hrs.
Rhizome:
Rinse in water | 3 min. peroxide | 35 min. bleach | 3 - 5 min. rinse | Soak in 5% PPM* for 2 hrs.
Media:
4 cups water | 1/2 teaspoon MS w/vitamins (50% MS) | 2 tablespoons sugar (25g/l)| 2 ml PPM | 2 ml BAP(1mg/ml) | PH 5.6
To tissue culture seeds leave out BAP but don't forget they have a stratification period. Stratify with GA3.
*Make the PPM with the same media your culturing with, but before adding sucrose and adjusting the PH. Make the same as 2% PPM but with more PPM
