Excision of the flower from the peach [Prunus persica (L.) Batsch.] flower bud raised the 50% injury temperature of flowers (cooled at 1C/hour) from -18 and -20C to -10 and -13C on two test dates, 26 Feb. 1988 and 5 Dec. 1990, respectively. Ice inoculation of the excised flowers at -3C further raised the 50% injury temperature to -7 and -8C for the two dates, respectively. These observations suggest that supercooling is a true mechanism for avoiding freezing injury. Low temperature scanning electron microscopy of freeze fractured cells verified that the flower froze intracellularly, whereas the subtending tissue froze extracellularly. Ice inoculation of the flower and the flower bud axis from which the scales were removed demonstrated that a barrier to ice propagation (effective to -11C) from the flower bud axis to the flower was present. This barrier may involve the provascular strands and a cell zone at the flower base (BZ) that were devoid of intercellular spaces. These tissues also had smaller cells, smaller vacuoles, greater ratio of cell wall thickness to cell size than tissue just below the BZ, which may result in greater cell rigidity and restriction of extracellular freezing. The cells outside the provascular strands at the base of the flower were also lacking in intercellular space, were smaller in size, and had a higher ratio of cell wall thickness to cell size compared to cells near the base of the scales. In the intact flower buds in which the flowers supercool below -11C, the presence of the first and second scales was important to full expression of supercooling because their removal raised the supercooling point, whereas the removal of lower scales did not. Sequestration of ice by the first two subtending scales during the early stages of freezing may be important to the creation of a dry region at the flower base that prevents ice propagation into the flower at temperatures below -11C.
Relationship between Phospholipid Breakdown and Freezing Injury in a Cell Wall-Less Mutant of Chlamydomonas reinhardii