Efficient regeneration of Sphagnum fallax from isolated protoplasts.

Authors:
Amla Batra, Horst Binding, Susanne Rasmussen, Hansjörg Rudolph, Georg H Waetzig
Year of publication:
2003
Volume:
39
Issue:
2
Issn:
1054-5476
Journal title abbreviated:
IN VITRO CELL DEV-PL
Journal title long:
In vitro cellular & developmental biology : journal of the Tissue Culture Association
Impact factor:
1.814
Abstract:
While the in vitro clonal propagation of peat mosses (Sphagnaceae) in bioreactors has been established since the late 1980s, it has never been possible to regenerate Sphagnum species from isolated protoplasts, which is a key step towards the production of closely defined genetically modified clones. The present study describes an efficient protocol for protoplast isolation and regeneration of Sphagnum fallax. Protoplast survival rates of over 50% and regeneration rates of up to 20% were achieved by using excised capitulum buds as starting material and by co-cultivating Sphagnum protoplasts with protoplasts from a chlorophyll-deficient Solanum hybrid clone. Besides the effects of nutrient components and differential osmotic readjustment of the regenerant cell clusters, the interference of unique Sphagnum phenolics, sphagnum acid and hydroxybutenolide, with protoplast isolation efficiency is demonstrated.