Auxin-polyamine interaction in the control of the rooting inductive phase of poplar shoots in vitro

Abstract

Changes in endogenous free IAA and its conjugate IAAaspartate as well as in endogenous polyamines were analysed in in vitro poplar shoots during their inductive phase, under the effect of the rooting auxin NAA, in combination or not with polyamines or inhibitors of polyamine biosynthesis. In vitro raised poplar shoots rooted 100% when treated by NAA (0.3 mg/l) for 7 h, the previously determined duration of the inductive phase. Spermidine and aminoguanidine (AG, an inhibitor of diamine oxidase) alone were unable to promote rooting under these conditions, and counteracted the NAA rooting effect. Putrescine and cyclohexylamine (CHA, an inhibitor of spermidine synthase) on the contrary did not oppose the NAA effect and promoted up to 40% rooting when applied alone. The levels of free indoleacetic acid (IAA) and of its aspartate conjugate IAAsp elevated up to peaks situated at the 7th and 8th h, respectively in the basal parts of the NAA-treated shoots. Putrescine, when added toegether with NAA, did not affect the typical IAA and IAAsp increases (except a displacement of the peaks to the 8th and 10th h, respectively); when applied alone, it provoked elevation of their levels to limited extents. Quite similar results were recorded with CHA. Spermidine and AG counteracted the NAA induced elevations of IAA and IAAsp. The level of endogenous putrescine typically increased up to a peak at the 6th h in the basal part of the NAA-treated shoots only, was slightly affected by exogenous putrescine and CHA application but significantly reduced by spermidine and AG supply. The NAA inducing rooting treatment did not affect the variation of the levels of endogenous spermidine and spermine but putrescine treatment (also spermidine to a lesser extent) led to increases of their levels. The relationships between auxin and putrescine temporary accumulation are discussed as well as the involvement of the different polyamines in the rooting inductive process.