Osmotic Stress-Induced Polyamine Accumulation in Cereal Leaves

Putrescine and spermidine accumulate in cereal cells and protoplasts exposed to various osmotica (sorbitol, mannitol, proline, betaine, or sucrose). The response is fast (1-2 hour lag), massive (50- to 60-fold increase in putrescine), and is not due to release of putrescine from a bound form or to conversion from spermidine. It rather involves the activation of the biosynthetic pathway mediated by arginine decarboxylase (ADC; EC 4.1.1.19) (Flores and Galston 1982 Science 217: 1259). Polyamine accumulation and the rise in ADC activity in osmotically stressed tissue are prevented by ADC inhibitors (alpha-difluoromethylarginine, d-arginine, and l-canavanine) but are not affected by alpha-difluoromethylornithine and methylornithine, inhibitors of the alternative putrescine biosynthetic enzyme ornithine decarboxylase (EC 4.1.1.17). Putrescine accumulation by oat and corn leaves is maximal in solutions only slightly hyperosmotic (0.4 molar). The stress response, which declines with leaf age, is completely prevented by cycloheximide (10 to 50 micrograms per milliliter) when added during the first hour of exposure to osmoticum, and partially by transcription inhibitors (cordycepin, Actinomycin D, 5 to 20 micrograms per milliliter). Oat seedlings allowed to wilt by withholding water also show a rise in polyamine titer and ADC activity. This response is not readily reversible upon rewatering.