The Metabolism of Hormones during Seed Germination and Dormancy: IV. The Metabolism of (S)-2-C-Abscisic Acid in Ash Seed.

Embryos from dormant and stratified Fraxinus americana seed were incubated with (S)-2-(14)C-abscisic acid (ABA) under a variety of conditions. Both dormant and stratified embryos rapidly metabolize abscisic acid to phaseic acid, dihydrophaseic acid, and an unidentified polar metabolite apparently derived from dihydrophaseic acid. Although the stratified embryos may have an increased capacity to metabolize abscisic acid, our calculations suggest that such an increased capacity would probably not be physiologically significant.Dormant intact seeds also metabolize (S)-2-(14)C-abscisic acid during stratification at 5 C or incubation at 25 C. The metabolites appear to be similar to those observed in excised embryos although by 12 days of stratification a fourth metabolite is observed. More than 90% of the (14)C-abscisic acid was metabolized after 26 days of stratification at 5 C or after 12 days of incubation at 25 C. Stratification at 5 C leads to the breaking of dormancy while incubation at 25 C does not.

The Metabolism of Hormones during Seed Germination and Release from Dormancy: III. The Effects and Metabolism of Zeatin in Dormant and Nondormant Ash Embryos.

Zeatin and zeatin-9, beta-ribonucleoside enhance the germination of dormant ash embryos. While the first macroscopic signs of germination appear only after about 72 hours, 12 hours of exposure to 50 mum zeatin is as effective as continuous incubation. There must be barriers against transport out of the embryos since 8-(14)C-zeatin and its metabolites, zeatin-9, beta-ribonucleoside, the 5'-mono and the suspected di- and triphosphates, accumulate against a concentration gradient. Zeatin ribonucleoside is about as effective as zeatin in enhancing embryo germination, yet the internal 8-(14)C-zeatin level is lower by a factor of about 50 when the ribonucleoside is fed. The physiological effects of zeatin and abscisic acid on the germination of ash embryos are antagonistic. There is, however, no evidence that abscisic acid has a significant effect on 8-(14)C-zeatin uptake or conversions.

Asymmetry, its importance to the action and metabolism of abscisic Acid.

Unlabeled and (14)C-labeled enantiomorphs of abscisic acid (ABA) were obtained through acetylcellulose chromatography and tested as inducers of abscission, as inhibitors of seed germination, and as antagonists of gibberellic acid-induced synthesis and release of alpha-amylase. The activity of the R isomer was either equal to or less than that of the naturally occurring S form. Greatest differences were in the inhibition of root-related growth. In excised beam axes, although uptake of S-[(14C)]ABA is faster, the internal concentration of R-ABA is higher because of faster conversion of S-ABA to inactive metabolic products. In axes a reversal in chirality is less important to the physiological action of ABA than to its metabolism.

Metabolically active glucosides in oleaceae seeds: I. The effects of germination, growth, and hormone treatments.

The seeds of six woody species of Oleaceae representing three genera, contain high concentrations of water-soluble glucosides, with major absorption maxima below 240 nanometers. In Fraxinus americana seeds three of these compounds, designated GL-3, GL-5, and GL-6, account for almost 10% of the dry weight. They are found in the endosperm and embryo but not in the pericarp. While the level of GL-5 is not particularly influenced by the physiological state of the embryo, that of GL-3 and GL-6 decreases as a result of germination and growth during a 10-day period. As the concentrations of GL-3 and GL-6 decrease, new ultraviolet-absorbing compounds are formed. The changes in the concentration of the ultraviolet-absorbing glucosides during cold temperature after-ripening, prior to germination, are small. When germination of dormant embryos is induced with gibberellic acid, the concentrations of GL-3 and GL-6 decrease in a manner similar to that observed with nondormant embryos. In the presence of abscisic acid no losses of GL-3 and GL-6 were observed. It is suggested that GL-3 and GL-6 fulfill some definite functions in the germination and growth of F. americana embryos, and that gibberellic acid and abscisic acid can exert a regulatory effect on the metabolism of these glucosides.

Abscisic Acid levels and seed dormancy.

Dormant seeds from Fraxinus species require cold-temperature after-ripening prior to germination. Earlier, we found that abscisic acid (ABA) will inhibit germination of excised nondormant embryos and that this can be reversed with a combination of gibberellic acid and kinetin. Using Milborrow's quantitative "racemate dilution" method the ABA concentration in 3 types of Fraxinus seed and pericarp were determined. While ABA was present in all tissues, the highest concentration was found in the seed and pericarp of dormant F. americana. During the chilling treatment of F. americana the ABA levels decreased 37% in the pericarp and 68% in the seed. The ABA concentration of the seed of the nondormant species, F. ornus, is as low as that found in F. americana seeds after cold treatment. Experiments with exogenously added ABA solutions indicate that it is unlikely that the ABA in the pericarp functions in the regulation of seed dormancy. However, the ABA in the seed does seem to have a regulatory role in germination.

Germination of Excised Fraxinus ornus Embryos With and Without Phleomycin.

In excised embryos of Fraxinus ornus, the first macroscopic sign of germination is a curvature near the root apex. Histological studies show that this curvature is due to cell elongation on the convex side and is accompanied by cell division. Metabolic changes, manifested by the disintegration of protein bodies and tissue differentiation, also occur during this time. In embryos treated with phleomycin cell division is completely inhibited but elongation, root hair formation, disintegration of protein bodies, and differentiation are still detectable. Phleomycin also drastically inhibits the formation of chlorophylls and the fresh weight increase of embryos.