The origin and transport of the IAA responsible for rooting was studied in carnation (Dianthus caryophyllus L.) cuttings obtained from secondary shoots of the mother plants. The presence of mature leaves in the cuttings was essential for rooting. Removal of the apex and/or the youngest leaves did not reduce the rooting percentage as long as mature leaves remained attached. Removal of mature leaves inhibited rooting for a 24-day period during which the basal leaves grew and reached maturity. After this period rooting progressed as in intact cuttings. Auxin (NAA + IBA) applied to the stem base of defoliated cuttings was about 60% as effective as mature leaves in stimulating rooting. Application of NPA to the basal internode resulted in full inhibition of rooting. The view, deduced from these results, that auxin from mature leaves is the main factor controlling the rooting process was reinforced by the fact that mature leaves contained IAA and exported labelled IAA to the stem. The distribution of radioactivity after application of (5-3H)-IAA to mature leaves showed that auxin movement in the stem was basipetal and sensitive to NPA inhibition. The features of this transport were studied by applying 3H-IAA to the apical cut surface of stem sections excised from cuttings. The intensity of the transport was lower in the oldest node than in the basal internode, probably due to the presence of vascular traces of leaves. Irrespective of the localization of the sections and the carnation cultivar used, basipetal IAA transport was severely reduced when the temperature was lowered from 25 to 4 degrees C. The polar nature of the IAA transport in the sections was confirmed by the inhibition produced by NPA. Local application of IAA to different tissues of the sections revealed that polar auxin transport was associated with the vascular cylinder, the transport in the pith and cortex being low and apolar. The present results strongly support the conclusion that IAA originating from the leaves and transported in the stem through the polar auxin transport pathway was decisive in controlling adventitious rooting.

(3)H-IAA transport in excised sections of carnation cuttings was studied by using two receiver systems for recovery of transported radioactivity: agar blocks (A) and wells containing a buffer solution (B). When receivers were periodically renewed, transport continued for up to 8 h and ceased before 24 h. If receivers were not renewed, IAA transport decreased drastically due to immobilization in the base of the sections. TIBA was as effective as NPA in inhibiting the basipetal transport irrespective of the application site (the basal or the apical side of sections). The polarity of IAA transport was determined by measuring the polar ratio (basipetal/acropetal) and the inhibition caused by TIBA or NPA. The polar ratio varied with receiver, whereas the inhibition by TIBA or NPA was similar. Distribution of immobilized radioactivity along the sections after a transport period of 24 h showed that the application of TIBA to the apical side or NPA to the basal side of sections, increased the radioactivity in zones further from the application site, which agrees with a basipetal and acropetal movement of TIBA and NPA, respectively. The existence of a slow acropetal movement of the inhibitor was confirmed by using (3)H-NPA. From the results obtained, a methodological approach is proposed to measure the variations in polar auxin transport. This method was used to investigate whether the variations in rooting observed during the cold storage of cuttings might be related to changes in polar auxin transport. As the storage period increased, a decrease in intensity and polarity of auxin transport occurred, which was accompanied by a delay in the formation and growth of adventitious roots, confirming the involvement of polar auxin transport in supplying the auxin for rooting.

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We describe here an experimental system to study the age-related decline of adventitious root formation in Arabidopsis thaliana L. (Heynh), ecotype Landsberg erecta (Ler). The system is based on the different rooting capacity of hypocotyls from de-rooted juvenile (12-day-old) and adult (26-day-old) plants. Hypocotyls from de-rooted juvenile plants rooted readily within a week of culture, and the rooting process was not dependent on exogenous auxin. In contrast, hypocotyls from de-rooted adult plants rooted poorly and only after longer periods of time. Exogenously applied auxin had no effect on rooting of hypocotyls from de-rooted adult plants. Rooting capacity, although correlated with the transition to flowering, did not depend on this transition. Root induction declined in a similar manner when the transition to flowering was delayed, either genetically with the fve mutant or physiologically with short days. The results showed that rooting of hypocotyls from de-rooted adult plants depended on the effect of peptides containing the RGD motif. Both the percentage of rooting and the number of roots were largely increased when the hypocotyls were treated transiently with the RGD peptide. The effect of the RGD peptide was a necessary, but not sufficient, condition for rooting of hypocotyls from de-rooted adult plants.

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