Cytokinin Reversal of Abscisic Acid Inhibition of Growth and α-Amylase Synthesis in Barley Seed

The effect of cytokinin, kinetin, on abscisic acid (dormin) inhibition of α-amylase synthesis and growth in intact barley seed was investigated. Abscisic acid at 5 × 10^?5M nearly completely inhibited growth response and α-amylase synthesis in barley seed. Kinetin reversed to a large extent abscisic acid inhibition of α-aniylase synthesis and coleoptile growth. The response curves of α-amylase synthesis and coleoptile growth in presence of a fixed amount of abscisic acid (6 × l0^?6M) and increasing concentrations of kinetin (from 5 × l0^?7M to 5 × 10^?5 M) showed remarkable similarity. Kinetin and abscisic acid caused synergistic inhibition of root growth. Gibberellic acid was far less effective than kinetin in reversing abscisic acid inhibition of α-amylase synthesis and coleoptile growth. A combination of kinetin and gibberellic acid caused nearly complete reversal of abscisic acid inhibition of α-amylase synthesis but not the abscisic acid inhibition of growth. The results suggest that factors controlling α-amylase synthesis may not have a dominant role in all growth responses of the seed. Kinetin possibly acts by removing the abscisic acid inhibition of enzyme specific sites thereby allowing gibberellic acid to function to produce α-amylase.     

Effets de l'acide gibbérellique et de la kinétine sur le développement de l'activitéα-amylasique durant la croissance de l'Orge

Effects of gibberellic acid and kinetic on α-amylase production during the germination of barley. - The action of gibberellic acid and kinetin, alone or combined at different concentrations, has been studied on α-amylase production in whole barley seedlings and in embryoless endosperms in course of the six first days of development in the dark. The classic activation of α-amylase synthesis by gibberellic acid has been confirmed both in whole seeds and in embryoless endosperms. Kinetin inhibits α-amylase synthesis after the third day of germination but has no effect on isolated endosperms. When gibberellic acid and kinetin are given simultaneously gibberellic acid stimulated during the three first days just as it does alone, kinetin inhibits after the third day also as it was alone so that the two regulators act, without interactions, at different stages in the time. These effects of kinetin are be independent. A critical examination of the techniques used in the literature in the stud of amylase is made.     

Selective effect of hormones on nucleic acid metabolism during germination of pear embryos

1. The effect of hormones on (32)P incorporation into various RNA fractions in germinating pear embryos was studied by fractionation on methylated albumin-kieselguhr columns. Abscisic acid inhibited labelling of soluble RNA, DNA-RNA hybrid and light-ribosomal RNA fractions with (32)P and this effect was reversed by both kinetin and gibberellic acid. 2. Kinetin reversed the inhibition by abscisic acid of (32)P incorporation into total ribosomal RNA and appeared to promote labelling of heavy-ribosomal RNA. Gibberellic acid was more active than kinetin in reversing the inhibition by abscisic acid of labelling of the DNA-RNA hybrid fraction with (32)P, but in contrast with kinetin appeared to increase further the inhibition by abscisic acid of labelling of total ribosomal RNA. 3. The percentage of radioactivity in various RNA fractions showed marked variation in response to hormones. 4. The pattern of labelling of RNA in pear embryos during reversal of inhibition by abscisic acid with a combination of kinetin and gibberellic acid was similar to that after cold-treatment of dormant pear embryos. This is suggestive of hormonal interplay in dormancy release by cold-treatment in pear embryos.     

A Comparative Study of the Changes in Root Growth, Induced by Coumarin, Auxin, Ethylene, Kinetin and Gibberellic Acid

The effect of coumarin, IAA, ethylene, kinetin and gibberellic acid on roots of maize and wheat was investigated. Sterile attached and detached roots and isolated elongation zones were used. In some experiments a semi-sterile procedure was followed. The effects of the different regulators separately or in various combinations together with coumarin were studied on the root growth with regard to division, elongation and swelling of the cells. The ethylene production in isolated elongation zones was measured after treatment with coumarin, IAA, PCIB, kinetin, colchicine and dinitrophenol. The results show the following: 1) Each substance produces a specific morphologic pattern. 2) Changes in polarity were demonstrated for auxin-induced swelling in cell divisions and cell expansion and for coumarin-induced swelling in cell divisions. Other cell expansion in swollen parts was due to cylindric cells increasing in width while retaining their cylindric form. 3) Coumarin-induced inhibition could not be counteracted by IAA, PCIB, carbon dioxide, kinetin, gibberellic acid or Cycocel. 4) The ethylene production in isolated elongation zones increases noticeably after kinetin treatment, less strongly after auxin treatment and least after coumarin treatment. The production of ethylene does not seem to be correlated with the morphogenetic effect of the different substances. 5) The isolated elongation zones reacted to a) IAA and kinetin with an increase in length in some cases and b) gibberellic acid with a reduction of their width. 6) The inhibitory effect of coumarin on the growth in length of the elongation zones was diminished by kinetin. The swelling produced by coumarin in these zones was reduced by gibberellic acid. The effects just mentioned of kinetin and gibberellic acid were considered as indirect ones. - From the present findings it was concluded that concomitant effects of auxin, ethylene, cytokinins and gibberellins are not obligatory for coumarin to exert its morphogenetic effects on root growth. In discussing the results some pitfalls in studies of growth reactions after application of hormones to roots containing meristem were emphasized.     

Effect of Growth Retardants on α-Amylase Production in Germinating Barley Seed

The effect of growth retarding compounds, (2-chloroethyl)trimethylammonium chloride (CCC), 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride (AMU-1618), tributyl-2,4-dichlorobenzylphosphonium chloride (Phosfon D) and N-dimethylamino succinamic acid (B-995) on α-amylase production in germinating barley seed was studied. Seeds were germinated in growth retardants in presence and absence of gibberellic acid (GA₃). CCC, AMO-1618 and Phosfon D inhibitedα-amylase production in germinating seed and the effect was reversed by GA₃ Phosfon D and AMO-1618 were stronger inhibitors of α-amylase production than CCC. CCC was by far the strongest inhibitor of all the other analogs tested. B-995 was comparatively only slightly inhibitory. The results reported here, when viewed in light of the results of other workers, provide good evidence that CCC, AMO-1618 and Phosfon D inhibit α-amylase production by inhibiting the synthesis of gibberellin or gibberellin-like hormone(s) during germination of barley seed. Consistent with other reports, B-995 possibly acts by other mechanism (s).     

The effect of selected plant hormones on in vitro proliferation of hyphae of Glomus fistulosum

Effects of N⁶-benzyladenine, kinetin, zeatin, N⁶-benzyladenosine, kinetin riboside, zeatin riboside, jasmonic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, abscisic acid and gibberellic acid on proliferation of hyphae of arbuscular mycorrhizal (AM) fungus Glomus fistulosum were studied under axenic conditions in vitro. The growth of intraradical hyphae of G. fistulosum was fully suppressed by 30 μM indole-3-acetic acid, but a perceptible decrease in the proliferation of the hyphae was observed already at 3 μM. Because such concentration is near the concentrations common in root tissues in vivo, the effect may be biologically significant. Similar effect was also observed for Glomus mosseae. Inhibitory effects of abscisic acid and cytokinins occurred only at very high, non-physiological concentrations. Ribosylated cytokinins showed stronger inhibition effects than their non-ribosylated counterparts. No stimulation of proliferation of hyphae by any plant hormone tested was observed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The hormonal regulation of bud outgrowth in Phaseolus vulgaris L.

Summary Aqueous solutions of indole acetic acid, kinetin, gibberellic acid and abscisic acid were applied singly and in combination to the decapitated stem stump of Phaseolus seedlings. Application of indole acetic acid will not completely replace the intact stem apex with regard to the inhibition of lateral bud extension. The greatest inhibition of bud growth is obtained when indole acetic acid is applied in combination with both kinetin and abscisic acid. Treatment with gibberellic acid causes massive bud growth even in the presence of indole acetic acid, kinetin and abscisic acid. Although both abscisic acid and kinetin have only a slight promoting effect on bud outgrowth when applied singly, these hormones will modify the effects of indole acetic acid and gibberellic acid.     

Involvement of plant growth substances in the alteration of leaf gas exchange of flooded tomato plants

Ethylene, abscisic acid, and cytokinins were tested for their ability to either induce or prevent the changes which occur in gas exchange characteristics of tomato (Lycopersicon esculentum Mill. cv. Rheinlands Ruhm) leaves during short-term soil flooding. Ethylene, which increases in the shoots of flooded plants, had no effect on stomatal conductance or photosynthetic capacity of drained plants. Abscisic acid, which also accumulates in the shoots of flooded plants, could reproduce the stomatal behavior of flooded plants when sprayed on the leaves of drained plants. However, photosynthetic capacity of drained plants was unaffected by abscisic acid sprays. Cytokinin export from the roots to the shoots declines in flooded plants. Spray applications of benzyladenine increased stomatal conductance in both flooded and drained plants. In addition, the decline in photosynthetic capacity during flooding was largely prevented by supplementary cytokinin applications. The possible involvement of these growth substances in modifying leaf gas exchange during flooding is discussed.     

Cytokinins as inhibitors of root growth

The elongation of roots of wheat ( Triticum aestivum L. cv. Diamant II), flax ( Linum usitatissimum L. cv. Concurrent) and cucumber ( Cucumis sativus L. cv. Favör) seedlings in the dark was strongly inhibited by various native and synthetic cytokinins (kinetin, benzyladenine, isopentenyladenine, zeatin and their corresponding 9-ribosides). An inhibition of 50% was obtained for wheat roots with 3 · 10⁻⁹ M zeatin and for flax roots with 6 · 10⁻⁹ M isopentenyladenine. The ribosides were in all cases less inhibitory. The inhibition was reversed by various types of 'antiauxins' and 'antiethylenes' (such as structural auxin analogues, uncouplers, specific inhibitors of ethylene synthesis, free radical scavengers, inhibitors of ethylene action). These substances as a rule counteract also inhibitions caused by auxins. Auxins and cytokinins stimulate ethylene production synergistically, and the similar inhibitory effects of these two types of hormone can be understood if it is assumed that their effect is at least partly mediated through ethylene. The cytokinins must be considered as possible natural inhibitors and regulators of root growth.     

Light-controlled Cycocel Reversal of Coumarin Inhibition of Lettuce Seed Germination and Root Growth

Lettuce seed germination or lettuce root elongation after germination in water was inhibited by coumarin and these inhibitions were reversed by Cycocel. 2.53 × 10³ M Cycocel reversed the inhibition of seed germination by 6.8 × 10⁴ M coumarin. and 6.32 × lO^?4 M Cycocel reversed the inhibition of root elongation by 3.4 × 10³ M coumarin. No other analogs of Cycocel reversed these coumarin induced inhibitions of growth. Cycocel reversal of coumarin inhibition of lettuce seed germination occurred in red light but not in far-red light or darkness. The red-far-red system was photoreversible. Cycocel and kinetin appear to act similarly in reversing coumarin inhibition of germination. Gibberellin A₃ and IAA were unable to reverse these coumarin induced inhibitions. A common mechanism is suggested for Cycocel reversal of coumarin and lAA inhibition of growth.     

Quantitative Relationship between Various Growth Substances and Bud Production in Funaria hygrometrica. A bioassay for abscisic acid

Eleven growth substances were tested for their bud producing capacity in Funaria hygrometrica. IAA and gibberellic acid showed no response, neither did inositol, abscisic acid and adenine. The cytokinins: benzylaminopurine, kinetin and mercaptopurine showed a quantitative relationship while methylaminopurine did not. When different amounts of abscisic acid were added together with 10 μg kinetin (which gave maximal production of buds), there existed a linear relationship between reduction in buds and abscisic acid added. Results are obtained in two weeks and it is therefore a fairly quick method for assaying certain cytokinin and also abscisic acid.     

Measurement of enzymes related to sucrose metabolism in permeabilized Chlorella vulgaris cells

Sucrose phosphate synthase (UDP-glucose: D-fructose-6-phosphate-2-glucosyl transferase, EC 2.4.1.14), sucrose synthase (UDP-glucose: D-fructose-2-glucosyl transferase, EC 2.4.1.13) and invertase (β-D-fructofuranoside fructohydrolase, EC 3.2.1.26) were measured in toluene permeabilized cells of Chlorella vulgaris Beijerinck. All three activities were detected at all stages of the growth curve; sucrose synthase and sucrose phosphate synthase showed a zone of maximum activity, while invertase increased with time of growth. Sucrose phosphate synthase and sucrose synthase (sucrose synthesis direction) were stimulated by divalent cations and inhibited by UDP. This inhibition could be reversed by Mg²⁺ or Mn²⁺. Sucrose phosphate synthase activity was inhibited by inorganic phosphate and was enhanced by glucose-6-phosphate, but was insensitive to sucrose. Arbutine decreased sucrose synthase activity in both directions. Sucrose cleavage was inhibited by divalent cations and by pyrophosphate. The effects on the enzyme activities of the presence of 2,4-dichlorophenoxyacetic acid (2,4-D), gibberellic acid, abscisic acid and kinetin in the growth medium were investigated. Sucrose synthase activity was practically unaffected by all plant hormones tested, except for the presence of kinetin which stimulated the activity. Sucrose phosphate synthase activity was increased by both kinetin and abscisic acid. The effect of the latter was partially reversed by the presence of gibberellic acid. 2,4-D and kinetin were potent stimulators of invertase activity.     

The effect of plant growth regulators on growth, morphology and condensed tannin accumulation in transformed root cultures of Lotus corniculatus

This study concerns the effects of four different classes of plant growth regulators on root morphology, patterns of growth and condensed tannin accumulation in transgenic root cultures of Lotus corniculatus L. (Bird's-foot trefoil). Growth of transformed roots in 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in decreased tannin levels relative to controls at concentrations of 10^-6 M and above, while gibberellic acid (GA₃) inhibited tannin accumulation at concentrations of 10^-7 M and above. Benzyladenine (BA) had little effect at low concentrations (10^-7 M and below) but resulted in an increase in tannin levels at 10^-6 M. Abscisic acid had little effect on levels of condensed tannins at any of the concentrations used. Experiments involving growth regulator addition and medium transfer demonstrated that 2,4-D inhibition of tannin accumulation could be reversed by GA₃ and BA, while GA₃ downregulation could only be reversed by the addition of 2,4-D. Although 2,4-D inhibited tannin accumulation, addition of 2,4-D to root cultures grown for 14 or 28 days in the absence of plant growth regulators stimulated both growth and tannin biosynthesis. Characteristic alterations in root morphologies accompanied growth regulator-mediated modulation of tannin biosynthesis. Growth in 2,4-D resulted in partially de-differentiated root cultures while growth in GA₃ produced roots with an elongated phenotype. Restoration of tannin biosynthesis in 2,4-D-treated roots was accompanied by root re-differentiation and the production of new lateral roots.Abbreviations ABA abscisic acid - BA benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid 3 - FW fresh weight     

Effect of plant hormones on sucrose uptake by sugar beet root tissue discs

Abscisic acid (ABA), auxins, cytokinins, gibberellic acid, alone or in combination were tested for their effects on short-term sucrose uptake in sugar beet (Beta vulgaris cv USH-20) roots. The effect of ABA on active sucrose uptake varied from no effect to the more generally observed 1.4-to 3.0-fold stimulation. A racemic mixture of ABA and its trans isomer were more stimulatory than ABA alone. Pretreating and/or simultaneously treating the tissue with K⁺ or IAA prevented the ABA response while cytokinins and gibberellic acid did not. While the variable sensitivities of beet root to ABA may somehow be related to the auxin and alkali cation status of the tissue, tissue sensitivity to ABA was not correlated with ABA uptake, accumulation, or metabolic patterns. In contrast to ABA, indoleacetic acid (IAA) and other auxins strongly inhibited active sucrose uptake in beet roots. Cytokinins enhanced the auxin-induced inhibition of sucrose uptake but ABA and gibberellic acid did not modify or counteract the auxin effect. Trans-zeatin, benzyladenine, kinetin, and gibberellins had no effect on active sucrose uptake. None of the hormones or hormone mixtures tested had any significant effect on passive sucrose uptake. The effects of IAA and ABA on sucrose uptake were detectable within 1 h suggesting a rather close relationship between the physiological activities of IAA and ABA and the operation of the active transport system.     

Interactions between Gibberellic Acid, Ethylene, and Abscisic Acid in Control of Amylase Synthesis in Barley Aleurone Layers

Gibberellic acid-induced α-amylase synthesis in barley (Hordeum vulgare L.) aleurone layers was inhibited by abscisic acid, and the inhibition was partly removed by additional gibberellic acid alone and by ethylene alone. Together additional gibberellic acid and ethylene almost eliminated abscisic acid inhibition of amylase synthesis. Time course studies of these phenomena showed that the effect of abscisic acid, ethylene, and varying concentrations of gibberellic acid on the course of amylase synthesis were either to speed up or slow down the whole process and not to affect the lag phase or the linear phase separately. The data are discussed in relation to previous studies of abscisic acid-gibberellic acid interaction.     

Cyclic Purine Mononucleotides: Induction of Gibberellin Biosynthesis in Barley Endosperm

The de novo synthesis of α-amylase in barley endosperm and isolated aleurone layers is induced by 3′,5′-cyclic purine mononucleotides and gibberellic acid. The induction of α-amylase by cyclic purine mononucleotides is prevented by 2,4-DNP, inhibitors of RNA and protein syntheses, CCC, AMO-1618 and phosfon. The induction of α-amylase formation by 3′,5′-cyclic purine mononucleotides, but not by gibberellic acid, is also blocked by inhibitors of DNA synthesis. Extracts from cyclic AMP-treated endosperm halves exhibit a characteristic gibberellin-like activity which is detectable within 12 hours from the addition of the cyclic AMP. On paper chromatograms this gibberellin-like activity is located at the Rf typical for GA₃. Its formation is prevented by inhibitors of DNA synthesis, CCC and AMO-1618. Glucose inhibits the formation of α-amylase induced by gibberellic acid. Glucose has no effect on the cAMP-induced gibberellin biosynthesis. The evidence shows that the cyclic purine mononucleotides induce DNA synthesis, which results in gibberellin biosynthesis, which in turn activates the synthesis of α-amylase.     

Variations de quelques aclivités enzymatiques (peroxydase, catalase, AIA-oxydase) et de la teneur en polyphénols au cours de la germination de I'Orge. Influence de la kinétine

Peroxidase catalase, IAA-oxidase and polyphenol content of growing barley coleoptile. Effect of kinetin. - Kinetin strongly inhibits root and coleoptile growth of germinating barley in the dark. Treated coleoptiles become senescent before the untreated ones. Soluble proteins content, peroxidase, catalase and IAA-oxidase activity were greatly increased in treated coleoptiles while the level of polyphenols was reduced. These biochemical effects joined with the other property of kinetin to diminish α-amylase synthesis in the endosperm are discussed in relation to growth and in connection with the classic view of a cytokinin retarded senescence.     

Interrelations between Gibberellic Acid, Cytokinins and Abscisic Acid in Retarding Leaf Senescence

The interrelation between the effects of abscisic acid (ABA) and the effects of cytokinins and gibberellic acid in retarding leaf senescence was investigated. Leaf discs from plants of Taraxacum megallorrhizon, Rumex pulcber and Tropaeolum majus were floated on solutions of cytokinin or GA to which given amounts of ABA were added. After five days, chlorophyll was extracted and the amount estimated spectrophoto-metrically. The interrelation between the effects of abscisic acid and cytokinins differed from that between the effects of ABA and gibberellic acid. Abscisic acid reduced the senescence retarding effect of GA more than that of cytokinins. A high concentration of cytokinins nullified the senescence enhancing effect of low concentrations of ABA. GA did not reverse the effects of ABA.     

Hormonal control of isoperoxidases in lentil embryonic axis

Detachment of the cotyledons from the lentil (Lens culinaris Med.) embryonic axis causes in the latter an increase in total peroxidase activity which is shown to be due to enhancement of specific cathodic isoperoxidases. Kinetin treatment of attached or detached axes promotes activity of essentially the same cathodic isoperoxidases. In addition kinetin enhances the activity of two anodic peroxidases and represses specifically that of a cathodic one. Abscisic acid inhibits the production of all isoenzymes in the presence or absence of kinetin. Cytokinin and abscisic acid actions are discussed in relation to the nature of a wounding hormone and the role of natural inhibitors in cotyledons during germination. Indoleacetic acid stimulates the activity of certain isoenzymes which are also stimulated by kinetin, whereas in others the effects of the two hormones are different. Specific inverse effects of indoleacetic acid and kinetin are demonstrated on the two most cathodic isoperoxidases. Indoleacetic acid-kinetin interactions on the cathodic isoperoxidases have been found in the literature and are discussed as a possible mechanism for explaining interactions of the two regulators on growth and other physiological processes.