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Environmental and Stress Physiology

Role of Abscisic Acid in the Induction of Freezing Tolerance in Brassica napus Suspension-Cultured Cells ¹

Department of Plant Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5, Institut de recherche en biologie végétale, Université de Montréal, 4101 rue Sherbrooke est, Montréal, Quebéc, Canada H1X 2B2

Brassica napus suspension-cultured cells could be hardened in 6 days at 25°C by the addition of mefluidide or ABA to the culture medium. Cells treated with mefluidide (10 milligrams per liter) or ABA (50 micromolar) attained an LT₅₀ of –17.5°C or –18°C, respectively, while the LT₅₀ for the comparable nonhardened control (sucrose) was –10°C. The increased freezing tolerance of mefluidide-treated cells was paralleled by a 4- to 23-fold increase in ABA, as measured by gas-liquid chromatography using electron capture detection. Application of 1 milligram per liter of fluridone, an inhibitor of abscisic acid biosynthesis, prevented the mefluidide-induced increase in freezing tolerance and the accumulation of ABA. Both these inhibitory effects of fluridone were overridden by 50 micromolar ABA in the culture medium. On the basis of these results, we concluded that increased ABA levels are important for the induction of freezing tolerance in suspension-cultured cells.

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