Auxin-gibberellin relationships in their effects on hypocotyl elongation of light-grown cucumber seedlings. Responses of sections to auxin, gibberellin and sucrose

The sensitivity of light-grown cucumber hypocotyl sections toIAA and GA₃ depends on the degree of aging of the tissue. Agreater response to GA₃ was obtained with young tissue, whilethat to IAA was obtained with relatively old tissue. The responseto IAA reached a maximum at about 15 hr of incubation; the youngerthe tissue the earlier the time of maximum response. The responseto GA₃ continued for more than 70 hr with a constant growthrate. Very young tissue started to respond to GA₃ without lagtime; the older the tissue the later the start of the response. Sucrose (2%) inhibited IAA-induced elongation, while there wasa distinct synergism between GA₃ and sucrose. The promotiveeffect of sucrose on GA₃-induced elongation was also obtainedwhen sections were pretreated with sucrose, then transferredto GA₃. Mannitol (1%) strongly inhibited IAA-induced elongation,but not GA₃-induced elongation. (Received December 6, 1972; )     

Auxin-gibberellin relationships in their effects on hypocotyl elongation of light-grown cucumber seedlings VI. Promotive and inhibitory effects of kinetin

The interaction of kinetin with IAA and GA₃ on the elongationof hypocotyl sections of Cucumis sativus L. cv. National Picklingwas studied. Kinetin in the concentration range of 10^–7M to 10^–4 M markedly inhibited IAA-induced elongation,while in a lower range from 10^–10 M to 10^–8 M, itsynergistically enhanced IAA-induced elongation. Kinetin alonein this range had no effect. A 5-to 15-min pulse treatment seemsenough to induce the maximum effect for both inhibition andpromotion. Since the magnitude of the maximum inhibition dependedon the concentration and not on the duration of treatment, thereaction in the cell caused by kinetin seemed to be completedwithin a short period. Washing of the sections with distilledwater after kinetin treatment (30 min) did not significantlyeliminate the kinetin effect. This probably indicates that thebinding of kinetin molecules to a supposed acceptor is not reversible.Interaction of kinetin with GA₃ in their pretreatment effectson IAA-induced elongation shows that in the inhibitory concentrationrange, the kinetin effect was partly overcome by GA₃, and thatin the promotive range, the magnitude of the enhancement wasdetermined by kinetin regardless of the presence of GA₃. Theeffect of kinetin seems to dominate over that of GA₃ indicatingthat the modes of their pretreatment effects differ from oneanother. (Received June 24, 1977; )     

Auxin-gibberellin relationships in. their effects on hypocotyl elongation of light-grown cucumber seedlings IV. Inhibition of IAA-induced elongation by N,N'-dicyclohexylcarbodiimide and its reversal by gibberellin A3

IAA-induced elongation and control growth of light-grown cucumberhypocotyl sections were markedly inhibited by DCCD, an inhibitorof membrane-bound ATPases. The concentration effective for inducingmarked inhibition was more than 10^–5 M. At 10^–5M DCCD, there was an apparent antagonism between IAA and DCCD.At 5 x 10^–5 M DCCD, the inhibition was partially recoveredby 10^–4 M of IAA. The results might indicate a close associationof the auxin action with membrane-bound ATPases. The DCCD inhibitionwas so strong that treatment with 10^–4 M DCCD for about5 min significantly suppressed further growth and longer incubationkilled the sections. In contrast, DCCD had not inhibitory effecton both control growth and IAA-induced elongation if GA₃ waspresent simultaneously. DCCD treatment followed by GA₃ treatmentstill resulted in the inhibition, suggesting that the inhibitionwas not reversible. In order to obtain reversal of DCCD inhibitionby GA₃ both compounds must be present at the same time. TheGA₃ effect is discussed in connection with the mechanism ofDCCD action on membrane-bound ATPases. (Received October 6, 1975; )     

Auxin-gibberellin relationships in their effects on hypocotyl elongation of light-grown cucumber seedlings III. Gibberellin specificity in its enhancing effect on IAA-induced elongation

Pretreatment effects of different gibberellins, helminthosporicacid, cyclic AMP and Kinetin on subsequent IAA-induced elongationwere tested in cucumber hypocotyl sections. Gibberellin A₇ wasmore active than GA₃, while gibberellin A₃ was almost inactive.Both helminthosporic acid and cyclic AMP mimicked GA₃-action,though the degree of their activity was less. Kinetin pretreatmentresulted in marked inhibition of IAA-induced elongation. Thepretreatment effect of GA₃ was also reflected in a greater responceof the sections to synthetic auxins. (Received October 6, 1973; )     

Actinomycin D Inhibits the GA3-Induced Elongation of Azuki Bean Epicotyls and the Reorientation of Cortical Microtubules

Actinomycin D inhibited the elongation of epicotyl segmentsfrom azuki bean seedlings that was induced by simultaneous treatmentwith IAA and GA₃. The drug also inhibited the elongation ofthe segments that was caused by IAA alone when it was appliedtogether with IAA. When the segments were pretreated with GA₃and then incubated with IAA, GA₃ promoted the elongation causedby IAA and brought about a predominance of transverse corticalmicrotubules (MTs) in the epidermal cells of the segments. Thechange in the arrangement of MTs caused by pretreatment withGA₃ was evident 1 h after the start of subsequent incubationwith IAA when the effect of pretreatment with GA₃ on the elongationhad not yet become apparent. Pretreatment with GA₃ did not causeany change in the arrangement of MTs when GA₃-pretreated segmentswere not incubated subsequently with IAA. Although actinomycinD applied before treatment with IAA did not inhibit the IAA-inducedelongation, the drug diminished the promotion of the elongationcaused by pretreatment with GA₃ and prevented GA₃ from bringingabout a predominance of transverse MTs when the drug was appliedduring the pretreatment with GA₃. GA₃-induced synthesis of mRNA seems to be involved in the promotionby GA₃ of IAA-induced elongation and in the GA₃-induced rearrangementof cortical MTs. (Received June 15, 1993; Accepted August 16, 1993)     

Control of Hypocotyl Hook Angle in Phaseolus mungo L. Role of Growth Substances

Effects of growth hormones on the hook angle and light responseof Phaseolus mungo L. hypocotyl hooks are described and theresults are discussed with reference to the functions of otherparts of the seedling in controlling the growth and shape ofthe hook. Apically applied IAA (indolyl acetic acid) prevented hook openingin decapitated seedlings in the dark and in all the red-irradiatedseedlings. [¹⁴C]IAA experiments showed that only a small quantityof IAA (2–6 ?g per hook) was required to produce theseeffects, and that transport of IAA through the hook was negligibleand unaffected by red irradiation. ABA (abscisic acid) had little effect on the hooks or theirlight response. Applied ethylene and IAA-induced ethylene slightly closed thehooks, but only slightly reduced light-induced opening. IAAreduced the effect of ethylene in the dark, but after irradiationthe hooks appeared more sensitive to the ethylene in the presenceof IAA, resulting in light-induced hook closure. Basally applied kinetin (6-furfurylaminopurine) prevented decapitatedhooks from opening in the dark, especially when GA₃ (gibberellicacid) was also present. Some combinations of kinetin and GA₃(with high kinetin concentrations) also prevented light-inducedopening, but combinations with lower kinetin concentrationsallowed almost as much opening as was found in intact hooks. It is proposed that the terminal parts act by regulating thesupply of cytokinins and gibberellins from the basal parts,and that IAA does not mediate this funotion in this species. The results are compared with those reported for other species.     

Enhancement of Gibberellic Acid-stimulated Hypocotyl Elongation of Lettuce (Lactuca sativa L. cv. Grand Rapids) by Phlorizin and Light

The interaction of light, gibberellic acid (GA₃), and phlorizinin the growth of lettuce (Lactuca sativa L. cv. ‘GrandRapids’) hypocotyls was investigated. At all concentrationsof GA₃, phlorizin enhanced GA₃-induced growth at luminous intensitiesabove 50 ft-c (continuous light). Without GA₃, phlorizin hadno effect on hypocotyl growth in the light but it inhibitedgrowth in the dark. Both seedlings and hypocotyl sections respondedto phlorizin in the presence of GA₃. There was no iteractionbetween phlorizin and KCl. Water-growth was severly inhibitedby light. GA₃,-induced growth was slightly inhibited by light,and then only at luminous intensities above 50 ft-c. Thus, relativeto H₂O-growth, GA₃-induced growth increased with increasingluminous intensity up to 450 ft-c, where it reached saturation.It seems that a synergism may exist between light and GA₃ aswell as between phlorizin and GA₃. Lactuca sativa L, lettuce, hypocotyl elongation, gibberellic acid, phlorizin, light     

Gibberellin-colchicine interaction in elongation of azuki bean epicotyl sections

In azuki bean (Azukia angularis = Vigna angularis) epicotylsections, gibberellin A₃ (GA₃) enhanced the growth promotingeffect of indoleacetic acid (IAA), but showed no growth effectwhen applied alone. Sections showed practically no cell division.The promoting effect of GA₃ on section growth seems to be dueto its promoting effect on cell elongation. The diameters of sections treated with IAA increased, but thediameters of sections treated with GA₃ together with IAA remainedconstant. GA₃ seems to suppress cell expansion in a directiontransverse to the cell axis. Colchicine at a concentration with no inhibiting effect on IAA-inducedelongation almost completely reversed the effect of GA₃ On the basis of these results, the participation of wall microtubulesin GA₃-induced elongation is discussed. (Received October 22, 1971; )     

Relationship of IAA-oxidase Activity to Gibberellinand IAA-induced Elongation of Light-grown Cucumber Seedlings

The growth and IAA-oxidase activity of light-grown cucumber seedlings (cv. Aonagajibae) were investigated in response to GA₃ and IAA. Both GA₃ and IAA induced significant elongation of the hypocotyl. The fresh and dry weights of the hypocotyl increased due to GA₃ or IAA treatment, whereas no significant change was observed in the cotyledons of GA₃-treated seedlings as compared with the controls. The fresh and dry weights of IAA-treated cotyledons were both lower than those of controls. Treatment with GA₃ or IAA resulted in retardation of IAA-oxidase activity in the hypocotyl and cotyledons. The degree of retardation was less in the cotyledons than in the hypocotyl. An inverse relationship was recognized between GA₃- or IAA-induced elongation and IAA-oxidase activity in the hypocotyl. The auxin-mediated mechanism for gibberellin action was discussed.     

Hormonal Regulation of Hypocotyl Elongation in Lactuca sativa L.: I. EVIDENCE AGAINST THE INVOLVEMENT OF GIBBERELLINS IN THE ELONGATION OF HYPOCOTYL IN SEEDLINGS GROWN IN THE DARK

The kinetics of extension induced by GA₃¹ in the hypocotyl ofintact seedlings of Lactuca sativa are similar in the dark andin the light, and differs fundamentally from the kinetics ofelongation in the dark without GA₃. Both in continuous lightand in the dark, GA₃-induced promotion starts 24 h after incubation.In the dark, even low concentrations of GA₃, which do not affectthe length measured after 6 d when the extension of hypocotylalmost ceases, remove the lag period of 48 h which precedesextension, and prolong the high rate of elongation. FollowingGA₃ supply the hypocotyl length in the dark and in the lightdoes not differ until 48 h; thereafter the rate of elongationin the light is less, so that the final length of the hypocotylis 40 per cent shorter than that of the dark-grown seedlingswithout GA₃. IAA supplied apically to light-grown seedlings induces a weakpromotion at a concentration of 1 mg l^–1 only. With anincreasing concentration of GA₃ supplied simultaneously, theconcentration of IAA inducing a significant promotion decreases.A combined supply of both these regulators, however, does notrestore the light-mediated inhibition of hypocotyl elongationcompletely. The maximum decrease in hypocotyl length induced by the growthretardants AMO-1618, CCC, and B-9 supplied from the beginningin the dark does not exceed 70 per cent. Saturating doses ofGA₃ supplied in combination with any one of the retardants compensateonly a fraction of the decrease. The results have been interpreted to show that native GAs arenot involved in extension growth in the dark.     

Interaction of a Brassinosteroid with IAA and GA3 in the Elongation of Cucumber Hypocotyl Sections

A synthetic brassinosteroid, 22,23(S,S)-homobrassinolide (hBR),was examined for its interaction with IAA and GA₃ in the elongationof hypocotyl sections of light-grown cucumber (Cucumis salivusL. cv. Aonagajibai) seedlings. hBR alone was less active thanIAA. Its optimal concentration was around 10 µM and thelowest effective concentration between 10 and 100 µM,which is more than 100 times higher than that of brassinolide.hBR was more active in sections from younger seedlings. Itsgrowth-promoting effect was negated or greatly reduced by inhibitorsof auxin-induced elongation such as p-chlorophenoxyisobutyricacid and kinetin. hBR acted synergistically with IAA and 2,4-Dbut not with GA₃ showing only an additive effect. Sequentialtreatment of sections with hBR and then with IAA also resultedin synergistic enhancement of auxininduced elongation, but whenthe order of treatment was reversed, hBR was inactive. The synergisticeffect was obtained with 1 h pretreatment with hBR and couldbe reduced by subsequent washing with water. There was no sequentialinteraction between hBR and GA₃. The synergistic pretreatmenteffects of hBR and GA₃ were simply additive to each other. Amembrane-bound ATPase inhibitor, dicyclohexylcarbodiimide, inhibitedthe hBR-induced elongation, but did not affect GA₃-induced elongation.The findings led to the conclusion that brassinosteroids enhanceauxin action and possess growth-promoting activity which isindependent of that of gibberellin. (Received November 9, 1984; Accepted February 18, 1985)     

Stimulation of auxin-induced elongation of cucumber hypocotyl sections by dihydroconiferyl alcohol. Dihydroconiferyl alcohol inhibits indole-3-acetic acid degradation in vivo and in vitro

The mechanism by which dihydroconiferyl alcohol (DCA) stimulatesindole-3-acetic acid (IAA)-induced elongation of cucumber hypocotylsections was studied. Although DCA did not affect the uptakeof IAA-5-³H by hypocotyl sections, the endogenous level of IAA-5-³Hin DCA-treated sections was much higher than in DCA untreatedones. IAA-5-³H in the incubation medium was degraded in thepresence of hypocotyl sections, and this degradation of IAAwas inhibited by DCA. An in vitro experiment with horseradishperoxidase revealed that DCA inhibited the IAA degrading activityof the oxidase, as did caffeic acid and ferulic acid. Theseresults suggested that DCA enhances IAA-induced cucumber hypocotylelongation by acting as an antioxidant of IAA. (Received June 4, 1975; )     

Biological activities of rishitin, an antifungal compound isolated from diseased potato tubers, and its derivatives

Rishitin, a norsesquiterpene alcohol, found in infected, resistantpotato-tuber tissue completely inhibited zoospore germinationand germtube elongation of Phytophthora infestans (MONT.) DEBARY at 10^–3M. There was little difference in sensitivityto rishitin among races of Phytophthora infestans. IAA-inducedelongation of Avcna coleoptile sections and GA₃-induced elongationof wheat leaf sections were also inhibited by rishitin. Theinhibition of IAA-induced elongation of Avena coleoptiles wasrelieved to some extent by increasing IAA concentration. However,little relief of the inhibition of GA₃-induced elongation ofwheat leaf sections was obtained by increasing GA₃ concentration.No plant injury was observed at this concentration of rishitin(10^–3M). Examination of a series of rishitin derivatives indicated thatthe hydroxyl group at C-3 is indispensable for antifungal activity.This activity was intensified by saturating the double bondbetween the rings of rishitin and/or that of the isopropenylgroup at C-7, though activity decreased when oxygenated functionalgroups were introduced into the side chain. Aromatization of the A ring did not lower biological activities.The antifungal activities of most rishitin derivatives almostparalleled their activities as plant growth retardants. However,some compounds without antifungal activity were active as growthretardants. ¹Studies on the phytoalexins (5). (Received August 14, 1968; )     

Effects of light on auxin-induced elongation of light-grown cucumber hypocotyl sections

The effect of light on IAA-induced elongation of light-growncucumber hypocotyl sections was examined. There was no differencein the optimal concentration of IAA between light and darknessand the biphasic pattern of IAA-induced elongation also wasobtained in both states. Elongation of the first phase was notaffected by light, but that of the second phase was influencedby the light condition of the first phase; dark incubation forthe first 3 hr resulted in a larger IAA-induced elongation inthe second phase. For the maximum IAA-induced elongation, atleast 3 hr of light exposure in the second phase was necessary.Simultaneously applied photosynthetic inhibitors, DCMU, CMUand o-phenanthroline, synergistically enhanced IAA-induced elongationin light, but not in the dark. They were not effective whenIAA treatment preceded the treatment with them. (Received April 3, 1978; )     

Short-term kinetics of elongation growth of gibberellin-responsive lettuce hypocotyl sections

The short-term kinetics of growth of the excised lettuce (Lactuca sativa L.) hypocotyl were characterized with respect to the effects of gibberellic acid (GA₃), indole-3-acetic acid (IAA), KCl and pH. A Hall-device-based, miniaturized, linear displacement transducer was developed to measure the growth of 2-mm hypocotyl sections with 1-m resolution. Following treatment with GA₃, a lag time of less than 10 min was typically followed by an increase in growth rate with two acceleration phases, reaching a final elevated rate within about 1 h. The kinetics of the response to GA₁, a mixture of GA₄ and GA₇, and GA₉ were similar to the response to GA₃. There was no response to IAA treatment either in the presence or absence of GA₃. KCl alone had no effect on the growth rate, but caused an increase in rate when added after GA₃, with a lag time of usually less than 1 h. Responses to pH changes had lag times of a few minutes in all cases. A shift from H₂O to pH 6 buffer inhibited growth, while a shift from H₂O to pH 4 buffer resulted in a transient increase to a rate comparable to that induced by GA₃. A shift from pH 6 to pH 5 caused an increase in growth rate, followed by a gradual decline to an H₂O control rate after more than an hour. The responses to GA₃ at pH 4 and pH 5 were similar to that found for addition of GA₃ to water controls.Abbreviations GA gibberellin - GA₃ gibberellic acid - GA₁, GA₄₊₇, GA₉ gibberellins A₁, A₄₊₇, A₉ - IAA indole-3-acetic acid     

Gibberellin response in lettuce hypocotyl sections

Excised lettuce (Lactuca sativa L.) hypocotyl sections retain the ability to elongate in response to gibberellic acid (GA₃) addition. In 48 hr at 30 C a GA₃-treated segment more than doubles while a control segment elongates less than 50%. Auxin has no detectable effect on this system. Sensitivity to GA₃ is not decreased by apex or root removal. Of the experimental variables tested, temperature, sucrose, and preincubation in water affect growth both with and without GA₃. Blue and far red light inhibit growth without GA₃; this inhibition is reversed by GA₃. Potassium chloride stimulates growth of illuminated sections treated with GA₃ but has no effect on control growth. When sections are incubated in the dark, KCl has a promotive effect on elongation.     

Combined Effect of Some Growth Regulators on Growth and Gametangial Formation in the Liverwort Riccia gangetica Ahmad

The present work deals with the effect of IAA-kinetin, IAA-GA³and GA³-ABA interactions on growth and gametangial formationin Riccia gangetica in vitro. Inhibitory effect of high concentrationof IAA on vegetative growth is overcome by the co-addition ofkinetin. The best response in terms of fresh and dry weightyields of thalli is obtained by a combination of 10^–5mol dm^–3 kinetin+ 10^–7 mol dm^–3 IAA. Interactionof IAA and kinetin has an additive effect on archegonial formation.Co-addition of IAA and GA³ decreases production of archegoniaand antheridia as compared to those produced in response toIAA and GA³ alone, respectively. Combination of GA³ and ABAreduces vegetative growth, as well as the number of anthendiaand archegonia. Key words: Riccia gangetica, growth, growth regulators, gametangial formation     

Relationship between IAA-induced elongation growth and plasma membrane NADH oxidase in soybean (Glycine max Merr.) hypocotyls

A relationship between the activity of NADH oxidase of the plasma membrane and the IAA-induced elongation growth of hypocotyl segments in etiolated soybean (Glycine max Merr.) seedlings was investigated. The plasma membrane NADH oxidase activity increased in parallel to IAA effect on elongation growth in hypocotyl segments. Actually, NADH oxidase activity was stimulated 3-fold by 1 u,M IAA, and the elongation rate of segments was stimulated 10-fold by 10 iM IAA. The short-term elongation growth kinetics, however, showed that the IAA-induced elongation of hypocotyl segments was completely inhibited by plasma membrane redox inhibitors such as actinomycin D and adriamycin, at 80 μM and 50 μM respectively. In addition, 1 mM actinomycin D inhibited the IAA-stimulated NADH oxidase activity by about 80%. However, adriamycin had no effect on NADH oxidase activity of plasma membrane vesicles. Based on these results, the plasma membrane redox reactions seemed to be involved in IAA-induced elongation growth of hypocotyls, and the redox component responding to IAA was suggested to be NADH oxidase.     

Effects of IAA on oxygen-sensitive growth and on hydroxyproline protein level in cell wall

Indole-3-asscetic acid (IAA) accelerated the incorporation ofradioactivity derived from ¹⁴C-proline into the SLS-insolublecell wall fraction only when the sections were exposed to lowoxygen concentrations. However, IAA showed no effect on theratio of hydroxyproline to proline incorporated into the SLS-insolublefraction in both 20% and 8% O₂-treated sections. The amountof hydroxyproline rigidly bound to the cell wall increased withincreasing IAA concentration in 8% O₂-treated sections, whilethat of the 20% O₂-treated ones decreased with IAA treatment. On the other hand, IAA increased the amount of ¹⁴C-labeled hydroxyprolineincorporated into the SLS-insoluble fraction of sections treatedwith cycloheximide, and their elongation was greatly inhibited. Based on the results that O₂ and IAA affect the auxin-inducedand the oxygen- sensitive growth differently, we suggest thatboth types of growth may antagonize each other in response tochanges in O₂ and IAA concentrations, resulting in balancedgrowth in the cell. (Received October 7, 1977; )     

Hormonal Regulation of Gametangial Formation in the Moss Bryum argenteum Hedw

Phytohormones such as auxins, cytokinins, gibberellins, andabscisic acid differentially affect gametangial induction inmale and female clones of Bryum argenteum. Both IAA and GA³increased the percentage of fertile gametophores in the maleclone, and inhibited the response in the female clone. GA₃ wasmore effective than IAA in eliciting the response in the maleclone. Cytokinins, on the other hand, increased the productionof fertile gametophores in the female clone, and inhibited itslightly in the male clone. The two cytokinins tested (kinetinand DMAAP) were almost equally effective for the female clone. An Interaction of IAA and kinetin nullified their individualinhibitory effects on the female and male clones, respectively.Cyclic AMP prevented the inhibitory effect of kinetin in themale clone; whereas, in the female clone, it stimulated theresponse elicited by kinetin. Abscisic acid (ABA) acted as ageneral inhibitor of vegetative growth and gametangial inductionin this moss. However, the inhibition of gametangial inductionwas greater in the female clone which is also more sensitiveto ABA than the male clone.