T,-H, 1994. The effect of H days oi niicroEravily on receneralion of inuicl plants (rom protoplasts, -Phvsiol. Plant, 92; 404^1 I,"The growih and development of protoplasts of rapeseed {Brii.\.\iia napu\ L. cv Line) and carrol (Daucus carora L, cv. Na\ona> were studied onboard the Space ShuUle 'Discovery' during an 8-da\ Internationa] Microgravity Laboraloiy (IML-1) mission in January !992. The flight e\pei'imenls were carried out in 'Biorack'. a fully controlled cell biological experimental facilit\. under microgravity conditions and in a l-;i,' centrifuge. Parallel experiments were performed in a 'Biorack' m(tdule on the ground. After retrieval, some samples \^ele subcultured on appropriate media and analysed for callus growih and regeneration to inlact plants. The remainder v\ere used for biochemical analysis. Samples fixed on board the Space Shuttle were kepi in 1 ^. f glutaraldehyde fixative al 4°C for ^-^ days for microscopy analysis after retrieval. Prcttctplasls expctsed to micrograviiy conditiitns showed u delay in cell wall synthesis. Cells were swctllen in appearance and fttrmed cell aggregates wilh (tnlv tew cells. Callus were obtained from protoplasts cultured under niicrograviu (FOg), (tn ihe !-.£,' cenlrifuge on board the shultle (Fig), under normal l-,i.' c(tndilions
The growth and development of protoplasts of rapeseed (Brassica napus L. cv Line) and carrot (Daucus carota L. cv. Navona) were studied onboard the Space Shuttle‘Discovery’during an 8‐day International Microgravity Laboratory [IML‐l) mission in January 1992. The Flight experiments were carried out in‘Biorack'. a fully controlled cell biological experimental facility. under microgravity conditions and in a l‐g centrifuge. Parallel experiments were performed in a‘Biorack’module on the ground. After retrieval, some samples were subcultured on appropriate media and analysed for callus growth and regeneration to intact plants. The remainder were used for biochemical analysis. Samples fixed on board the Space Shuttle were kept in l% glutaraldehyde fixative at 4°C for 3–7 days for microscopy analysis after retrieval. Protoplasts exposed to microgravity conditions showed a delay in cell wall synthesis. Cells were swollen in appearance and formed cell aggregates with only few cells. Callus were obtained from protoplasts cultured under microgravity (Fogl). on the l‐g centrifuge on board the shuttle (Flg), under normal l‐g conditions on the ground (G1g) and on a centrifuge on the ground giving 1.4 g (Gl.4g). Regeneration of intact rapeseed plants was obtained from Flg. Glg and G1.4g. However, no plants were regenerated from protoplasts exposed to microgravity (Fog). Biochemical analysis indicated that the microgravity samples (Fog displayed a reduced packed cell volume, an increased concentration of soluble proteins per cell, and a reduced specific activity of peroxidase in the cytoplasm. Morphometric analysis of fixed samples demonstrated that 3‐day old protoplasts under microgravity conditions were significantly larger than protoplasts kept on the l‐g centrifuge in space. UItrastructural analysis by transmission electron microscopy showed that protoplasts exposed to microgravity conditions for 3 days had larger vacuoles and a slightly reduced starch content compared to Flg cells. Cell aggregates formed under microgravity conditions (Fog) had an average of 2–I cells per aggregate while aggregates formed under Flg had 8–12 cells.
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