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; )     

Biological activity of phyllosinol, a phytotoxic compound isolated from a culture filtrate of Phyllosticta sp.

1. Phyllosinol is a phytotoxic metabolite of Phyllosticta sp. Thissubstance at 100 µg/ml produced dark grey necrotic lesionson the leaf of red clover. Sensitivities of various plant speciesto phyllosinol differed both quantitatively and qualitatively.
2. Phyllosinol reduced root growth in rice seedlings by 60% at10^–4 M, whereas stimulation of root elongation occurredat a concentration range from 10^–9 to 10^–5 M.
3. Phyllosinolat 2.5x10^–4M promoted adventitious root formationin epicotylsof Azukia cuttings by about 100%. Promotion waspartly reducedby simultaneous application of cysteine.
4. IAA-induced elongationof isolated Avena coleoptile sectionswas inhibited by phyllosinolat a concentration range from 10^–5to 10^–3M.
5. Sulfhydrylcompounds, i.e. cysteine and glutathione relievedinhibitioncaused by phyllosinol in IAA-induced elongation ofAvena coleoptilesections.
6. GA₃-induced elongation of wheat leaf sections wasslightly inhibitedby phyllosinol at 10^–4M.
7. Phyllosinolalso has antibiotic activity. Among the organismstested, Phycomycetesand Gram-negative bacteria appeared mostsusceptible to phyllosinol.

(Received April 21, 1970; )     

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; )     

Factors Influencing Alanine Aminotransferase Activity in Leaves of Lolium temulentumL.: II. EFFECTS OF GROWTH REGULATORS AND PROTEIN BIOSYNTHESIS INHIBITORS

Effects of kinetin (K), gibberellin A₃ (GA₃), and 2-(chloroethyl)-trimethylammoniumchloride (CCC) on levels of alanine aminotransferase (GPT) andrates of protein synthesis were studied with both intact plantsand isolated leaf segments of Lolium temulentum L. In intactplants CCC stimulated and CA₃ reduced GPT activity, the effectsbsing much greater in 8.h than in 16-h photoporiods. CCC showedmaximum stimulatory effects at 10^–2 M and K at 5 x 10⁵M. No effect of GA₃ could be demonstrated with concentrationsup to 10^–4M. Both K and CCC retarded GPT decline in leafsections, the latter without associated effects upon pigmentbreakdown. Cycloheximide was highly effective in reducing proteinsynthesis in leaf sections. A close correlation between rateof protein synthesis and GPT activity was found over an inhibitorconcentration range from 10^–6 to 10^–4 M. The resultsare discussed in terms of possible methods of in vivo regulationof GPT activity.     

Auxin-gibberellin relationships in their effects on hypocotyl elongation of light-grown cucumber seedlings II. Effect of GA3-pretreatment on IAA-induced elongation

IAA-induced growth of light-grown cucumber hypocotyl sectionsis markedly enhanced by GA₃-pretreatment of the sections; thereis a distinct synergism between IAA and GA₃. Water pretreatmentalso enhances IAA-induced growth. On the other hand, IAA-pretreatedsections showed practically no further growth in response topost treatment with GA₃. The enhancing effect of GA₃ is obtainedwith only 30 min pretreatment, the maximum effect occuring with2 hr pretreatment. Pretreatment longer than 8 hr is less effective.This enhancing effect of GA₃ can be observed soon after posttreatment with IAA. The response of GA₃-pretreated sectionsto IAA is greater in pretreatment with higher concentrationsof GA₃, and higher degrees of synergism between IAA and GA₃are obtained at IAA concentrations less than 10^-4 M. This synergisticinteraction between GA₃ and IAA is more marked in aged hypocotylsections than in young sections. From these results we concludedthat gibberellin sensitizes hypocotyl cells to the subsequenteffect of auxin on cell elongation. (Received October 6, 1973; )     

Involvement of cellulose synthesis in actions of gibberellin and kinetin on cell expansion. Gibberellin-coumarin and kinetin-coumarin interactions on stem elongation

In azuki bean (Azukia angularis = Vignia angularis) epicotylsections, 5 ? 10^–4 M coumarin inhibited the incorporationof radioactivity from [U^–14C]glucose into the cellulosefraction by 35% in the absence of indole-3-acetic acid (IAA)and by 40% in the presence of 1 ? 10^–4 M IAA. There wasno inhibitory effect on the incorporation of radioactivity intothe other fractions. Coumarin at 5 ? 10^–4 M reversed thepromoting effect of 1 ? 10^–5 M gibberellin A₃ (GA) andthe inhibitory effect of 1 ? 10^–5 M kinetin on IAA-inducedelongation of sections with no significant effects on IAA-inducedelongation. Neither GA nor kinetin had any appreciable effectson cellulose synthesis. No inhibition of cellulose syntheiswas observed with 1 ? 10^–3 M colchichine, which has beenreported to have effects similar to those of coumarin on GA-or kinetin-affected stem elongation. Coumarin at 5 ? 10^–4M was ineffectual in breaking up wall microtubules, while adisrupting effect on wall microtubules was clearly demonstratedwith 3 ? 10^–4M colchicine. From these results, the possible involvement of cellulose synthesisin cell expansion controlled by GA or kinetin was suggested. (Received August 3, 1973; )     

Effects of Plant Growth Regulators on Grain-filling and Yield of Rice

Effects of three phytohormones (IAA₁, GA₃ and kinetin) on grain-fillingand the pattern of ³²P translocation from individual leavesto grains were studied at intervals of 7 days during the progressof reproductive development of rice (Oryza saliva L. cv. Jaya).The plants were sprayed with 100 µg ml^–1 aqueoussolutions of the hormones at 100 days, when the plants wereentering the reproductive stage. Kinetin produced a pronouncedeffect on grain-filling as well as on ³²P mobilization fromindividual leaf to grains and increased yield, possibly by increasingleaf longevity. GA₃ and IAA also increased the grain-fillingand ³²P mobilization significantly over control but the effectswere less marked than those of kinetin. Oryza sativa L., rice, grain yield, translocation, growth regulators, gibberellic acid, indol-3-yl acetic acid, kinetin     

Effects of Auxin and Gibberellin on Elongation of Young Hyphae in Neurospora crassa

IAA, 2,4-D and GA₃ promoted the elongation of young hyphae inNeurospora crassa at the optimum concentrations of 10^–6,10^–6 and 10^–4 M, respectively. The effects of IAAand GA₃ were additive. (Received June 17, 1983; Accepted December 22, 1983)     

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)     

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; )     

Effects of Gibberellic Acid and Naphthaleneacetic Acid on Petiole Senescence and Subtended Peduncle Growth of Cyclamen persicum Mill

Removal of the blade from the leaf subtending the first flowerbud on Cyclamen persicum ‘Swan Lake’ plants causedthe petiole of that leaf to senesce, but had no effect on thegrowth of the flower peduncle in the debladed petiole's axil.A 10 mg NAA l^–1 application generally had no effect onpetiole senescence, peduncle elongation or flowering date whenapplied to the cut end of the petiole after blade removal. A25 mg GA₃ l^–1 application or a combination of 25 mg GA₃l^–1 application or a combination of 25 mg GA₃ l^–1plus 10 mg NAA l^–1 delayed petiole senescence and enhancedpeduncle elongation and subsequent flowering. No treatment significantlyaltered peduncle length at the time of flowering. Cyclamen persicum Mill, ‘Swan Lake’, tissue receptivity, flowering, GA₃, NAA     

Effects of Auxin and Gibberellin on Conidial Germination and Elongation of Young Hyphae in Gibberella fujikuroi and Penicillium notatum

In Gibberella fujikuroi and Penicillium notatum, IAA, 2,4-Dand GA₃ promoted conidial germination and the elongation ofyoung hyphae. The promotive effects of IAA and GA₃ were additive.In both fungi, the concentrations of endogenous auxin and gibberellinin the culture media were 10^–10 to 610^–12M. (Received April 27, 1985; Accepted August 12, 1985)     

Inhibition of Gibberellic Acid-Induced Elongation-Growth of Pea Epicotyls by Xyloglucan Oligosaccharides

The biological activity of a cell wall-derived xyloglucan nonasaccharide(XG9) was investigated using a bioassay with entire pea epicotyls(Pisum sativum cv. Progress). The xyloglucan fragment was foundto inhibit gibberellic acid-induced elongation of etiolatedpea epicotyls with maximum inhibition at concentrations rangingfrom 10^–11 to 10^–9M. Growth of etiolated epicotylsin the absence of exogenously applied GA₃ was also inhibitedby XG9 in the same concentration range. A cell wall-derivedheptasaccharide (XG7) lacking the fucosyl-galactosyl-side chainshowed no inhibitory effect in the pea epicotyl bioassay withand without exogenous GA₃. Furthermore, the biological activityof a synthetic pentasaccharide (XG5), containing the fucosylgalactosyl-sidechain which is necessary for the biological activity was investigatedin the same bioassay. Compared to XG9 the pentasaccharide hada similar inhibitory activity on GA₃-promoted elongation aswell as on the endogenous growth in the absence of exogenouslyapplied GA₃, but did not exhibit a distinct concentration optimum. Key words: Elongation-growth, gibberellic acid (GA₃), oligosaccharides, pea, XG9     

Effects of Some Growth Regulators and Benzoic Acid Derivatives on Flower Initiation and Root Elongation of Pharbitis nil, Strain Kidachi

Seedlings of Pharbitis nil, strain Kidachi, were grown undercontinuous light at 20°C in vessels containing 5,000-mlnutrient solution, 24 plants per vessel. NAA (0.005–0.5µM), GA₃ (0.1–0.5 µM), kinetin (0.5–5µM), benzyladenine (0.05–5 µM) or abscisicacid (4 µM) added to the nutrient solution induced long-dayflowering, and the flowering was always accompanied by suppressionof root elongation. 3,4-Dichlorobenzoic acid (0.05–10µM) and some other benzoic acid derivatives which arehighly effective for the induction of flowering in Lemna paucicostataalso showed similar effects. Neither NAA, kinetin nor 3,4-dichlorobenzoicacid applied via the apical part of the hypocotyl could causeflowering or suppression of root elongation. Thus, the flower-inducingeffect of the above substances was presumed to be secondaryto the suppression of root elongation. Ethrel (1–50 µM)added to the nutrient solution suppressed root elongation, butdid not induce flowering probably because it has flower-inhibitingactivity. ¹ This paper is dedicated to the memory of Dr. Joji Ashida,the first president of the Japanese Society of Plant Physiologists. (Received December 15, 1982; Accepted February 25, 1983)     

Dormancy in Dioscorea: Generality of gibberellin-induced dormancy in asexual dormant organs

Effects of GA₃ and CCC application on the sprouting of bulbilsor subterranean dormant organs of 10 species in the genus Dioscoreawere observed. Although the efficiency of both chemicals differedby species, in general GA₃ inhibited and CCC promoted the sproutingof the above dormant organs. In some species, however, dilutedGA₃ (0.003–0.3 µM) has a promotive and diluted CCC(3–30 µM) has an inhibitive effect on sprouting. Effects of GA₃ application on shoot elongation were tested onsprouted bulbils. GA₃ promoted elongation when applied directlyto the shoots and inhibited it when applied to the bulbous parts. These results suggest that GA activates two opposing reactions—sprouting-promotingand sprouting-inhibiting—in these organs. The complicatedrelation between GA₃ or CCC concentrations and sprouting wereexplainable by assuming that the two counteractive reactionswere activated by GA in different degrees. ¹ Present address: Department of Biology, Faculty of Science,Yamagata University, Yamagata 990, Japan. (Received June 21, 1976; )     

Abscisic Acid and Apical Dominance in Phaseolus coccineus L. The Role of Tissue Age

Abscisic acid (ABA) applied to the decapitated second internodeof runner bean plants enhanced outgrowth of lateral buds onlywhen internode stumps were no longer elongating. Applied toelongating internodes of slightly younger plants, ABA causesinhibition of bud outgrowth. Together with 10^–4 M indol-3-ylacetic acid (IAA), ABA stimulated internode elongation and interactedadditively in the inhibition of bud outgrowth. A mixture of10^–5 M ABA and 10^–6 M gibberellic acid (GA₃ ) causedsimilar effects on internode growth as IAA + ABA, but was mutuallyantagonistic in effect on growth of the lateral buds. Abscisic acid, apical dominance, gibberellic acid, indol-3yl acetic acid, Phaseolus coccineus, bean     

Effects of a Plant-Growth Regulator, Prohexadione, on the Biosynthesis of Gibberellins in Cell-Free Systems Derived from Immature Seeds

Inhibition of the biosynthesis of gibberellins by prohexadione,3,5-dioxo-4-propionylcyclo-hexanecarboxylic acid, was studiedwith cell-free systems derived from immature seeds of Cucur-bitamaxima, Phaseolus vulgaris and Pisum sativum. Prohexadione,at a concentration of 10–⁴ M, inhibited C-7 oxidationof GA₁₂-aldehyde, C-20 oxidation of GA₁₅, conversion of C₂₀-gib-berellinsto C₁₉-gibberellins, 3ß-hydroxylation, 2,3-dehydrogenationof GA²⁰, 2,3-epoxidation of GA₅ and 2ß-hydroxylationof GA₉ and GA₂₀. The 3ß-hydroxylase activity appearedto be more sensitive to prohexadione than were the C-20 oxygenaseand the 2ß-hydroxylase activities. The conversionof mevalonic acid to GA₁₂-aldehyde and the 13-hydroxylationof GA₁₂ were not affected by prohexadione at a concentrationof 3 ? 10^–4 M. All of the steps inhibited by prohexadioneare oxidation steps catalyzed by soluble enzymes that require2-oxoglutarate, Fe²⁺ and oxygen, and all of them occur distalto the synthesis of GA₁₂-aldehyde in the biosynthesis of gibberellins. (Received April 4, 1990; Accepted September 14, 1990)     

Uptake of Gibberellin Methyl Esters by Spores and Induction of Dark Germination in Lygodium japonicum

In the fern Lygodium japonicum, the effect of the exogenousapplication of two gibberellin methyl esters, gibberellin A₄methyl ester (GA₄Me) and gibberellin A₂₀ methyl ester (GA₂₀Me)on spore germination in the dark and uptake of GA₄Me and GA₂₀Meby spores was investigated. Tritiated GA₄Me and GA₂₀Me wereprepared and used as radioactive tracers. The activity of GA₄Mewas more than 100-fold that of GA₂₀Me for the induction of sporegermination. When treated for 24 h, the activity for inducingspore germination remained after removal of the gibberellinmethyl esters from the medium. The amount of GA₄Me taken upby spores was more than three times that of GA₂₀Me throughoutthe 24 h time course of treatment. The uptake of both gibberellinmethyl esters was proportional to the external concentrationfor the range of concentrations between 10^–9 M and 10^–6M. When treated with the tritiated gibberellin methyl estersat 10^–6 M and 10^–7 M for 24 h, most of the gibberellinmethyl esters taken up by the spores were not metabolized. Althoughthe uptake of the two gibberellin methyl esters differed by3- to 5- fold, their abilities to induce spore germination differedby more than 100-fold. Therefore, the difference in the activityof the two gibberellin methyl esters regarding the inductionof spore germination could not be explained solely by the differencein their uptake. (Received January 11, 1988; Accepted May 26, 1988)     

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     

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.