Abstract: | The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 104M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar Alaska, an ABA-deficientmutant wil, and its wildtype. Freezinginjury was determined graphically as the temperature that caused50% injury (T50) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T50 for root tissuewas 1C for all genotypes, regardless of treatment. For Alaskashoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by 1.5 to 4.0C, while coldtreatment increased the freezing tolerance by 7.5 to14.8C. Cold treatment increased the freezing toleranceof shoot tips by 9 and 15C for wiland wild-type, respectively. Cold acclimationincreased endogenous ABA concentrations in Alaskashoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of Alaska, respectively.In Alaska shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in wiland wild-type shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization |