ISHS Acta Horticulturae 560: IV International Symposium on In Vitro Culture and Horticultural Breeding
ESTABLISHMENT OF A SEMICONTINUOUS BIOREACTOR CULTURE OF PHYSCOMITRELLA PATENS FOR MASS PRODUCTION OF PROTOPLASTS |
| Authors: | A. Hohe, G. Schween, R. Reski |
| Keywords: | ammonium tartrate, flowcytometric analysis, moss, suspension culture |
| Abstract:
For large scale protoplast transformation of the moss Physcomitrella patens (Hedw.) B.S.G. semicontinuous protonema suspension cultures in bioreactors were established and optimised with regard to the yield of protoplasts. Supplementation of Knop medium with 2.5 mM ammonium tartrate markedly improved the protoplast yield (7.4 x 104 protoplasts/mg dry weight). Bioreactor culture was performed in 5 l vessels. By harvesting on average 1100 ml/day which corresponds to a dilution rate of 0.22/d a semicontinuous culture was maintained for 49 days yielding 51 l of suspension culture. Beginning eleven days after the start of the bioreactor run around 6 % of the cells were polyploid (4C) which might indicate aging of the culture caused by phytohormone accumulation. | |
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