The role of gibberellins in the growth of floral organs of Pharbitis nil

Evidence that the synthesis of GA₃ is involved in the growthof floral orga'ns of Pharbitis nil is presented. GAs in floralorgans at different developmental stages were surveyed usingTLC followed by the bioassay with two dwarf rice seedlings,‘Tanginbozu’ and ‘Waito-C’. The amountof GAs in the petal and stamen increased rapidly after the petalemerged from calyx, reached a maximum 12 hr before anthesis,then declined markedly thereafter. The GA content in the calyxremained unchanged before and after anthesis, and that in thepistil increased after anthesis. Pharbitis flowers containedat least two active GAs, one of which was probably GA₃, theother appeared to be GA₁₉. GA₃ was detected in relatively largeamounts in both the petal and stamen during their rapid elongation.In the calyx, which showed little increase in fresh weight duringrapid flower growth, GA₉ was the dominant GA. Exogenously suppliedGA₃ promoted elongation of sections in excised young filaments.Sucrose was necessary for definite growth promotion by GA₃.GA₁₉ had little effect on filament elongation, and IAA was ratherinhibitive. (Received July 29, 1972; )     

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)     

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)     

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; )     

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; )     

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; )     

Total and Polar Lipid Biosynthesis during Crotalaria juncea Linn. Pollen Tube Growth: Effect of Gibberellic Acid, Indoleacetic Acid and (2-Chloroethyl)-phosphonic Acid

Gibberellic acid (GA₃) at 58 µM, indoleacetic acid (IAA)at 29 µM, and (2-chloroethyl) phosphonic acid (Ethephon)at 70 µM promoted pollen tube growth in Crotalaria junceapollen suspension cultures both in water and basal medium. GA₃stimulated [ l-¹⁴C]acetate incorporation into total lipids inboth media, whereas IAA enhanced incorporation in water culturesonly. On the contrary, Ethephon reduced the label in total lipidswhen supplemented in basal medium. Based on [l-¹⁴C lacetateincorporation into different phospho- and glycolipids, it isproposed that these growth regulators have a definite role inthe biosynthesis of lipid components of the membranes.     

Inoculation with Azospirillum lipoferum Affects Growth and Gibberellin Status of Corn Seedling Roots

Maize (Zea mays L., hybrid Cargill 147) seedlings, grown inaseptic conditions, were inoculated with three strains of Azospirillumlipoferum (Al op 33, Al iaa 320, and ATCC 29708) or culturedin different concentrations of indol-3-acetic acid (IAA) orgibberellin A₃ (GA₃). After 48 h, root length, root surfacearea, root dry weight, and shoot dry weight were measured inall treatments. Gibberellin content was evaluated in selectedroots of control and Azospirillum inoculated seedlings by gaschromatography-mass spectrometry-selected ion monitoring withthe use of deuterated gibberellins as internal standards. Thethree strains of A. lipoferum, IAA (2 ng ml^–1), and GA₃(40 to 400 pg ml^–1) significantly enhanced root growth.Improvement of root hair growth and density was obtained mainlywith A. lipoferum strain Al op 33 and GA₃ 40 pg ml^–1.GA₃ was identified by gas chromatography-mass spectrometry (byboth, full scan and selected ion monitoring) in a free acidfraction from roots of the seedlings inoculated with A. lipoferum.In the roots of the non inoculated seedlings GA₃ was found afterhydrolysis of a fraction expected to contain glucosyl conjugates. (Received April 26, 1993; Accepted September 27, 1993)     

In Vitro Phosphorylation of Proteins in IAA-Treated Mesocotyls in Zea mays

Five-mm sections of elongation zones of Zea mesocotyls wereincubated for designated periods with various concentrationsof IAA. In vitro protein phosphorylation in the soluble fraction(85,000 x g supernatant) prepared from the sections was analyzedby sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The phosphorylation of proteins in the soluble fraction thathad been prepared from sections incubated for 20 min in thepresence of 10{small tilde}^s M IAA was greater than that inthe sections incubated for 20 min without IAA. The amount ofphosphorylation of proteins per protein became higher when higherconcentrations increased (10{small tilde}⁸—10{small tilde}⁵M).The growth of sections incubated in the presence of 10{smalltilde}⁸ M IAA or higher concentrations was greater than thatof sections incubated in the absence of IAA. The promotion ofgrowth by IAA was greater at higher concentrations of IAA. Proteinsin the soluble fraction, prepared from sections incubated for20 min in the presence of 10{small tilde}⁵ M IAA, were phosphorylatedin the presence of either 10 fM cAMP, 10 µM cGMP, 100µM W-7, 100 µM W-5, 20 µM H-7 or 20 µMHA1004. The calmodulin antagonist, W-7, and the inhibitor ofprotein kinase C, H-7, inhibited the phosphorylation of proteinsstimulated by incubation with IAA. These results suggest thatIAA promotes cell elongation via protein phosphorylation thatdepends on calmodulin-dependent protein kinase and protein kinaseC. (Received November 29, 1995; Accepted May 20, 1996)     

A lack of correlation between the biological activity and rate of metabolism of ent-(3H)-17-kaurenoic acid by seedlings of dwarf rice cv. Tan-ginbozu

Significant leaf sheath elongation occurred within 24 hr afterapplication of 10 µg (0.67, µCi) of ent-(³H)-17-kaurenoicacid (KA) to individual seedlings of dwarf rice cv. Tan-ginbozu,but this growth was unaccompanied by production of significantlevels of radioactivity in more polar, acidic, ethyl acetate-solublemetabolites of (³H)-KA. However modest levels of radioactivityappeared in the highly water-soluble fraction by hour 24, subsequentto the most rapid phase of KA-induced growth. Growth continuedand by hour 48 was accompanied by the appearance of small amountsof radioactivity in polar, acidic products. It would appearthat KA per se, and not its metabolic products, may be responsiblefor the leaf sheath elongation noted at hour 24. On the speculation that it might be a metabolite of KA, gibberellinA₁₄ (GA₁₄) was applied simultaneously with (³H)-KA to individualrice seedlings. Several changes in the metabolism of ³H-KA inthe presence of GA₁₄ were noted, and GA₁₄ antagonized the KA-inducedsheath elongation. ¹Present address: Botany Department, Rhodes University, Grahamstown,6140, South Africa. ²Present address: Crop Science Department, University of Saskatchewan,Saskatoon, Sask. S7N OWO, Canada. (Received May 12, 1975; )     

Effects Produced on Tomato Plants, Lycopersicum esculentum, by Seed or Root Treatment with Gibberellic Acid and Indol-3yl-Acetic Acid

Growth in lengths of tomato stems and leaves was acceleratedby 5.0 µg gibberellic acid (GA₂) applied to the seed,or by 5.0, 0.5, and 0.05 µg given to the roots. Treatmentwith 5.0 µg also decreased bud number and lengthened thetime between bud appearance and fruit formation on the firsttruss by 1–8 days. Smaller amounts applied to roots shortenedthis time by 1–4 days. Indol-3yl-acetic acid at 0.5 µghad no effect, nor was simultaneous application of GA₃ and IAAto the roots more effective than GA₃, alone. Single applicationsof very small amounts of GA₃ to seeds or seedling roots thusproved capable of changing growth-rates of stems, leaves, andtrusses.The effects of treating tomatoes with GA₂ and with culturesof Azotobacter chroococcum, which contain small amounts of GA₃,and IAA, are compared.     

Gibberellin-Induced Changes in the Water Absorption, Osmotic Potential and Starch Content of Cucumber Hypocotyls

The effects of GA₃ on the water absorption, osmotic potentialand starch content of light-grown cucumber (cv. Aonagajibai)hypocotyl sections were examined. GA₃ (100µM.) stimulatedfresh weight increase as well as elongation. It had no effecton the dry weight change in the presence, or absence, of 50mM sucrose although dry weight increased significantly in thepresence of sucrose. In the absence of sucrose the weight wasunchanged. The osmotic potential of the epidermal cells of sectionsincubated with GA₃ was lower than that of the control, and thedifference between the two values was larger in young seedlings.When sections were incubated with sucrose, the osmotic potentialgreatly decreased. This decrease was more marked in the presenceof GA₃. GA₃ reduced the starch content of the sections bothin the presence and absence of sucrose. The total amount ofstarch, however, was markedly increased in its presence. Thedegradation of starch formed in advance from exogenous sucrosein light was not accelerated by GA₃. We discuss a possible rolefor gibberellin in cell elongation, based on our results, interms of cell water relations. (Received January 18, 1983; Accepted July 19, 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; )     

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; )     

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     

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 by Light of Hypocotyl Elongation and Levels of Endogenous Gibberellins in Seedlings of Lactuca sativa L.

Four 13-hydroxygibberellins, gibberellin A₁ (GA₁), 3-epi-GA₁,GA₁₉ and GA₂₀ were identified by full-scan GC/MS in extractsof lettuce seedlings (Lactuca sativa L. cv. Grand Rapids). Theresults suggest that the early-13-hydroxylation biosyntheticpathway to GA₁ functions in the lettuce seedlings. It was alsofound that GA₁ is active per se in the control of hypocotylelongation in lettuce seedlings. To investigate the relationshipbetween control by light of hypocotyl elongation and levelsof endogenous GAs in lettuce, endogenous levels of GAs werequantified by radioimmunoassay in seedlings that had been grownfor 5 days in the dark (5D) and in those that had been grownfor 4 days in the dark and then under white light for 1 day(4D1L). The endogenous level of GA₁ in the upper and lower partsof hypocotyls in 5D seedlings was about four times higher thanthat in 4D1L seedlings. The response of explants (hypocotylsegments with cotyledons) from dark-grown seedlings to GA₁ isknown to be similar in the dark and under white light when theexplants are treated with inhibitors of the biosynthesis ofGA. Therefore, the above information suggests that the highlevel of GA₁ in hypocotyls of dark-grown seedlings is responsiblefor the rapid elongation of hypocotyl, while irradiation bywhite light decreases the endogenous level of GA₁ in the hypocotylswith a resultant decrease in the rate of hypocotyl elongation. (Received March 13, 1992; Accepted May 21, 1992)     

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; )     

Effects of Fluorogibberellins on Plant Growth and Gibberellin 3r{beta}-Hydroxylases

3rß-Fluorogibberellin A₉ (3rß-fluoro-GA₉),3rßfluoro-GA₂₀, 3rß-fluorodeoxygibberellinC (3rß-fluoro-DGC) and 13-fluoro-GA₉ were prepared,and their effects on plant growth and gibberellin (GA) 3rß-hydroxyIaseswere examined. 3rß-Fluoro-GA₉ and 3rß-fluoro-GA₂₀promoted the growth of dwarf rice (Oryza sativa L. cv. Tan-ginbozu)seedlings to three times higher than the control seedlings ata dosage of 3 µ plant^–1, and 3rßfluoro-DGCto twice higher at the same dosage. 3rßg-Fluoro-GA₉was active in cucumber (Cucumis sativus L.) hypocotyl assay,its activity being about one-thirtieth as much as that of GA₄.3rß-Fluoro-GAs were active per se in promoting theshoot elongation of rice. 3rß-Fluoro-DGC inhibitedthe 3rß-hydroxylation of [³H₂]GA₉ to [³H]GA₄ by GArß-hydroxylase from bean (Phaseolus vulgaris L.),but 3rß-fluoro-GA₉ and 3rß-fluoro-GA₂₀ didnot show any effects on the enzyme activity. These 3rß-fluoro-GAsalso showed no or only a weak inhibitory effect on the rß-hydroxylasefrom pumpkin (Cucurbita maxima L.). 13-Fluoro-GA₉ promoted growthof rice and cucumber seedlings, and inhibited the 3rß-hydroxylasesfrom both bean and cucumber. 13-Fluoro-GA₉was converted into13-fluoro-GA₄ and 2,3-didehydro-13-fluoro-GA₉, in a cell-freesystem from bean, and conversion of 13-fluoro-GA₉ into 13-fluoro-GA₄was also observed in a cell-free system from pumpkin. Theseresults suggest that 13-fluoro-GA₉ is one of the substratesof GA 3rß-hydroxy-lases, and that 13-fluoro-GA₉ isactive as a result of the conversion to 13-fluoro-GA₄ in riceand cucumber seedlings. (Received October 27, 1997; Accepted March 13, 1998)     

Physiological Controls on the Production of Cleistogamous and Chasmogamous Flowers in Lamium amplexicaule L

Plants of Lamium amplexicaule, grown under short-day field conditionsin Northern California produce predominantly closed flowers.Under long-day field conditions, plants may produce up to 50per cent open flowers, the same total number of flowers beingproduced under each daylength regime. A 20 ml drop of GA₃ or GA₇ (100µM) was applied to themain shoot apex of plants growing under short-day conditions,and all subsequently produced flowers opened. CCC, an inhibitorof gibberellin synthesis, was applied (0.06 per cent) to thesoil of seedlings grown under long-day conditions. The CCC-treatedplants were dwarfed and bore only closed flowers. With GA₇ appliedexogenously to the CCC-treated shoots, inhibition was released,resulting in elongated internodes and open flower production.The timing of flower production and internodal extension inLamium amplexicaule are positively correlated. When floral primordiawere removed from main shoots, the average internode lengthsdecreased. The number of nodes produced in treated plants wasincreased as a result of flower removal. Exogenous GA₇ (10 µM)applied to the nodes from which flowers were removed resultedin internodal extension but had no effect on node number. Twoprocesses that may contribute to the control of the productionof open and closed flowers in Lamium amplexicaule are: (1) anincreasing anther sac size from lower to upper node flowersthat may exert control locally, via GN production, on corollaexpansion, and (2) a photoperiodic control. Lamium amplexicaule L, henbit, cleistogamy, chasmogamy, gibberellins, gibberellic acid (GA₃), (2 chloroethyl) trimethylammonium chloride (cycocel)