Peroxidase catalyzes the oxidation of indole-3-acetic acid.The primary products of this reaction stimulate growth in plants. Therefore, our concept is that an increase in peroxidase activity will increase the effect of indole-3-acetic acid as a growth hormone. Our objective was to study the effect of 2,3,5-triiodobenzoic acid, a growth regulator, on isoperoxidases in the cell wall and cytoplasm of Nicotiana. Isoperoxidases from the cell wall and cytoplasmic fractions were separated by acrylamide gel electrophoresis. We found that 2,3,5-triiodobenzoic acid and indole-3-acetic acid increase peroxidase activity in the cell wall. Since both 2,3,5-triiodobenzoic acid and indole-3-acetic acid increase the activity of the same isoperoxidase, we conclude that 2,3,5-triiodobenzoic acid synergizes rather than antagonizes auxin action, and we suggest that this increase in indole-3-acetic acid oxidase activity sensitizes plant tissues to auxin.Plant growth is inhibited by TIBA' and it also causes epinasty of leaves and stems (7,21).This "formative" type of growth inhibition resembles that caused by high concentrations of auxin. Nevertheless, data in the literature indicate that TIBA antagonizes the action of auxin (4). It is suggested that TIBA blocks auxin transport (4) or immobilizes auxin and thus renders it nonfunctional (2). Experiments of Bouck and Galston (3) show, however, that the auxin, blocked by TIBA, is functional and that TIBA stimulates growth caused apparently by endogenous auxin that accumulated above the area where TIBA was applied. Thimann and Bonner (19) also reported that TIBA augments the auxin effect in the pea and A vena tests.Our objective was to study the effect of TIBA on the peroxidase system in Nicotiana and relate these findings to the concept that the oxidation of IAA catalyzed by peroxidase in plants yields products that stimulate growth (13). We treated Nicotiana with TIBA and isolated cell wall and cytoplasmic peroxidases. Cell wall peroxidases were isolated from exudates of intact tissues. Cytoplasmic peroxidase enzymes were isolated from tissues devoid of cell wall peroxidase. Both extracts were subjected to electrophoresis on polyacrylamide gels. We found that both TIBA and IAA stimulated peroxidase activity in the cell wall but not in the cytoplasm. We postulate that an increase in peroxidase activity caused by TIBA may block auxin transport and, at the same time, increase the sensitivity of TIBA-treated tissues to auxin.'Abbreviation: TIBA: 2,3, 5-triiodobenzoic acid.MATERIALS AND METHODS Nicotiana tabacum L. cv. Xanthi-nc was grown for 7 to 8 weeks in the greenhouse and then sprayed with 1% polyoxyethylene sorbitan (20) monolaurate (Tween 202), or with 106 to 1O-3 M TIBA dissolved in 1% Tween 20. Leaves and apices were collected 1 to 2 days later and assayed for peroxidase. Cell wall peroxidases were separated from the cytoplasmic fraction by the vacuum infiltration method (1).To extract cell wall enzymes, we removed the midrib of each leaf and placed the leaf halves in a desi...