Preparatory studies for the use of plant protoplasts in space research

An experiment using plant protoplasts has been accepted for the IML-1 mission to be flown on a space shuttle in 1991. Preparatory experiments include studies of cell wall formation, cell division, the effect of simulated weightlessness using fast and slow rotating clinostats, and the development and testing of hardware for the IML-1 mission. After 24 h at 25 degrees C, protoplasts isolated from hypocotyls or leaves of rapeseed seedlings, or from carrot suspension cells, show 60, 20 and 15% cell wall formation, respectively. The time course of formation of the cell wall and cell division could be delayed by treatment at low temperatures or immobilization in alginate or agarose. This aspect is of importance in connection with problems of late access to the space shuttle before launch. At 4 degrees C only 18% of the rapeseed hypocotyl protoplasts had formed cell walls after 24 h. Protoplasts immobilised in agarose or alginate gradually regain their cell division capacity and after 72 h the frequencies are 51 and 26%, respectively, compared to non-immobilised control protoplasts. A significant decrease in cell division activity is observed after rotation for 6 h on the slow clinostat. A similar effect is not observed on the fast clinostat. Protoplasts, cultured in the specially designed plant chamber for up to 14 days established cell aggregates which have further developed into plants.