Persistent Supercooling and Silica Deposition in Cell Walls of Palm Leaves

Summary Leaves of Trachycarpus fortunei from palms growing in warm temperate regions of Europe and Japan as outdoor ornamentals were cooled down to low subfreezing temperatures, and the process of ice formation was followed using differential thermal analysis (DTA). Inactivation of photosynthetic function was determined by in vivo chlorophyll fluorescence, and the progress of injuries during freezing was assayed by viability tests after thawing. Ultrastructure of mesophyll cells was investigated by transmission electron microscopy. Silica incrustation of cell walls was documented by means of qualitative energy dispersive X-ray analysis (EDAX) on samples of Trachycarpus fortunei, Washingtonia filifera, and of the bamboo Pseudosasa japonica. DTA revealed typical freezing patterns, with a high temperature exotherm at about -6 to -8°C and a broad low temperature exotherm rising from about -13 to -14°C. As soon as the tissues froze, characteristic changes appeared in the kinetics of in vivo fluorescence. No irreversible inactivation of photosynthesis or damage was detectable at temperatures above appearance of the low exotherm. Within a short time after attainment of the exotherm peak, the leaf segments were found to be lethally damaged. Both of in vivo fluorescence and viability tests clearly indicated that Trachycarpus leaves remain in the persistent supercooled state down to quite low temperatures. The tensile state of water at threshold supercooling temperatures of Trachycarpus leaves was calculated according to the equation of Rajashekar and Burke (1982) and found to be about -14 MPa. An explanation for the permanent maintenance of such a large displacement from thermodynamic equilibrium could be that silica incrustation, as revealed by EDXA, favours supercooling by increasing the rigidity of the cell walls and/or resistance to the growth of ice crystals.