Rubbing-induced lignification in Bryonia dioica internodes is significantly impaired by N(o-hydroxyphenyl) and N(o-aminophenyl) sulfinamoyl tertiobutyl acetate, specific inhibitors of cinamyl alcohol dehydrogenase, an enzyme which is strictly associated with lignin monomer synthesis. Along with the reduction of lignification, these inhibitors counteract the inhibition of elogtion due to rubbing. These results indicate that lignification participates in the thigmomorphogenetic growth response of Bryonia dioica internodes. In a general way, the data point to the causal role of lignification in the limitation of plant growth.Rubbing young internodes of many plants results in growth cessation and increased radial expansion of the internodes (3, 5-9, 15). In Bryonia dioica the reduced elongation has been related to changes in cell wall mechanical properties resulting from an accelerated lignification (9).Several molecules able to reduce lignification have already been defined. Phenylalanine ammonia-lyase is inhibited by aaminooxy-,3-phenylpropionic acid (1) and cinnamate 4-hydroxylase by l-aminobenzotriazole (16). Unfortunately, these two target enzymes are involved in general phenylpropanoid metabolism rather than specifically associated with lignification. In addition, phenylalanine ammonia-lyase inhibitors act also on pyridoxal phosphate enzymes involved in ethylene (10) and amino acid synthesis (2). In contrast, the reductive enzyme cinnamyl alcohol dehydrogenase is strictly associated with the synthesis oflignin monomers and is inhibited in a highly specific way by OHPAS2 and NH2PAS (4,13 The last growing internodes (± 15 mm long) were gently rubbed after 1 d inhibitor feeding. For all analyses internodes were harvested 24 or 48 h after the rubbing treatments. All experiments were repeated at least three times (results ± SE).Lignin Content. Lignin content of cell wall preparations was estimated according to the technique reported before (9).
RESULTSRubbing-induced inhibition of internode elongation is significantly counteracted by treatment ofthe plants with OHPAS and NH2PAS, both supplied at the concentration of 40 ,uM (Fig. 1A).Inhibitor feeding at the concentration of 80 AM appeared toxic because of wilting of the plants within 24 h. The slight reducing effect of the inhibitors on elongation of nonrubbed internodes may be a thigmomorphogenetic response due to manipulation ofthe plants. This seems to be confirmed by a concomitant slight increase in lignin content (Fig. 1B). No significant differences