The temperature boundary for phase separation of membrane lipids extracted from Nerium oleander leaves was determined by analysis of spin label motion using electron spin resonance spectroscopy and by analysis of polarization of fluorescence from the probe, trans-parinaric acid. A discontinuity of the temperature coefficient for spin label motion, and for transparinaric acid fluorescence was detected at 7'C and -3°C with membrane lipids from plants grown at 45°C/32°C (day/night) and 20'C/15°C, respectively. This change was associated with a sharp increase in the polarization of fluorescence from trarns-parinaric acid indicating that significant domains of solid lipid form below 7'C or -3°C in these preparations but not above these temperatures. In addition, spin label motion indicated that the lipids of plants grown at low temperatures are more fluid than those of plants grown at higher temperatures.A change in the molecular ordering of lipids was also detected by analysis of the separation of the hyperfine extrema of electron spin resonance spectra. This occurred at 2'C and 33°C with lipids from the high and low temperature grown plants, respectively. According to previous interpretation of spin label data the change at 29'C (or 33°C) would have indicated the temperature for the initiation of the phase separation process, and the change at 7'C (or -3°C) its completion. Because of the present results, however, this interpretation needs to be modified.Differences in the physical properties of membrane lipids of plants grown at the hot or cool temperatures correlate with differences in the physiological characteristics of plants and with changes in the fatty acid composition of the corresponding membrane lipids. Environmentally induced modification of membrane lipids could thus account, in part, for the apparently beneficial adjustments of physiological properties of this plant when grown in these regimes.The evergreen higher plant Nerium oleander (oleander) acclimates to both hot summer and cool winter conditions and grows actively in these contrasting regimes (2 prevailing temperature is probably an important adaptive mechanism for this and other desert evergreen species, such as Larrea divaricata (9) and A triplex lentiformis (12), which experience large seasonal differences in environmental temperature. The variation in fatty acid composition ofA. lentiformis with growth temperature (I I) suggests that changes in the physical properties of membranes might be the basis for the acclimation.To characterize the oleander membrane lipids, we have utilized ESR4 spectroscopy and measurements of fluorescence intensity and polarization with the probe, trans-parinaric acid (25). Both techniques revealed the same temperature for the separation of solid and fluid phase lipids but indicate a need to reinterpret some previous ESR studies of plant membrane lipids. Furthermore, the results show that the physical properties of N. oleander membrane lipids change during thermal acclimation.
MATERIALS AND METHODSRooted cut...