Treatment of etiolated pea (Pisum sativum L.) internode tissue with ethylene gas inhibits elongation and induces lateral expansion. Precise kinetics of the induction of this altered mode of gro4vth of excised internode segments were recorded using a double laser optical monitoring device. Inhibition of elongation and promotion of lateral expansion began after about 1 hour of treatment and achieved a maximum by 3 hours. Similar induction kinetics were observed after treating internodes with coichicine and 2,6-dichlorobenzonitrile, an inhibitor of cellulose synthesis. In sealed flask experiments, ethylene had no detectable effect on incorporation of label from '4Cqglucose into any of the classical pectin, hemicellulose, or cellulose wall fractions. Ethylene inhibited fresh weight increase (total cell expansion) of both excised internode segments (in sealed flasks) and intact seedlings. Ethylene treatment resulted in an increase in cell sap osmolality in those tissues (intact and excised) which are inhibited by the ps. A model for ethylene-induced inhibition of elonption and induction of lateral expansion is presented.Ethylene inhibits elongation and induces lateral expansion of etiolated pea internode tissue (2, 3). Ethylene is thought to affect the mode of growth by altering microtubule and cellulose microfibril orientation (1,9,12,21,26). Colchicine and other agents which affect microtubule and microfibril orientation also inhibit elongation and induce lateral expansion (6). DCB3, a specific inhibitor of cellulose synthesis (14), induces lateral expansion in soybean tissue (30). Coumarin, a less specific inhibitor of cellulose synthesis, has been shown to induce lateral expansion in several kinds of tissues (7,10).Although the inhibition kinetics of elongation with ethylene treatment have been studied in detail (16)
MATERIALS AND METHODSSeeds of Pisum sativum L. (var Alaska) were soaked in tap water for 6 h, planted in vermiculite moistened with 0.1 mM CaCl2 in deionized H20, and grown for 7 d in darkness at 22C. For experiments using excised internode segments, a 1-cm section of third internode tissue was cut immediately below the apical hook. Unless otherwise stated, all excised tissues were incubated, 10 pieces/lot, in sealed 125-ml flasks with 10 ml of medium with or without 10 gl/l ethylene. The medium contained 5 mM K-phosphate (pH 6.8), 2% sucrose (w/v), 5 AM CoC12, and 1 gM IAA. Growth ofsegments in length and width was measured using the LOLA described by Taiz and Metraux (27). The measuring system was calibrated with a micrometer and the magnifications (x 118 for length and 598 for diameter) were found to be constant within the range used for these experiments. The SD among replicate experiments averaged less than 10% of the increase in growth of the segments. The tissue chamber and the mechanical lever system of the LOLA were enclosed in a sealed Plexiglas box with a volume of approximately 300 L. The atmosphere within this box was constantly circulated. The tissue segment in the LOLA chamber...