Lipid transition temperatures for rough and smooth microsomal membranes isolated from bean (Phaseolus vulgaris) cotyledon tissue at various stages of germination were determined by wide angle x-ray diffraction. The transition temperatures were established by recording diffraction patterns through a temperature series until a sharp x-ray reflection centered at a Bragg spacing of 4.15 A and denoting the presence of crystalline lipid was discernible. For rough and smooth microsomes from 2-day-old tissue, the transitions occurred at 0 C and 3 C, respectively, indicating that at this early stage in the germination sequence the membrane lipid is entirely liquid-crystalline at physiological temperture. By the 4th day of germination, the transition temperatures had increased to 32 C for smooth microsomes and 35 C for rough microsomes, indicating that at 29 C, which was the growth temperature, portions of the membrane lipid were crystalline. During the later stages of germination, the transition temperature for smooth microsomes continued to rise through 44 C at day 7 to 56 C at day 9, by which time the cotyledons were extensively senescent and beginning to abscise. There was also a dramatic increase in the proportion of membrane lipid in the crystalline phase at 29 C. By contrast, the rough microsomes showed little change in transition temperature and only a dight increase in the proportion of crystalline lipid during this late period in germination. The data indicate that substantial amounts of the lipid is senescing membranes are crystalline even at physiological temperature. Moreover, there is a temporal correlation between the appearance of this crystality and loss of membrane function, suggesting that the two may be causally related.The onset of senescence in cotyledons of germinating Phaseolus vulgaris is marked by a general metabolic decline which includes deterioration of membrane function. Plasma membranes, for example, exhibit decreased cation-sensitive ATPase and cholinesterase activities as well as a reduced capability for ATP-dependent cation transport (12,14). This loss of function correlates temporally with age-dependent alterations in the protein complement of the membrane (15). The enzymic activities of microsomal membranes also decline as senescence intensifies, and the levels of microsomal phospholipid phosphate decrease indicating that there is a structural disassembly of these membranes (18).There is increasing evidence that the physical state of membrane lipid is an important factor in determining such key membrane functions as enzyme activity and permeability. By means of Arrhenius plots it has been demonstrated for certain membrane-bound enzymes that temperature-induced changes in activation energy correlate with phase transitions in the membrane lipid (5, 6, 9, 25). In addition, the permeability of lipid vesicles and natural membranes can be altered by factors known to influence membrane fluidity (4,24,31 The rough microsomes were further purified by centrifugation through 1.8 M sucrose ...