Changes in Endogenous Gibberellin Levels in Tulipa Bulblets during Ontogeny

The endogenous gibberellin activity of Tulipa gesneriana cv.Apeldoorn bulbiets of field-grown mother bulbs was determinedduring ontogeny by paper and gas-liquid chromatography and thedwarf pea bioassay. It was shown that the gibberellin activityof bulblets increased dramatically in November-March and declinedsharply in April-July. The increase in gibberellin-like substanceswas considered to be derived primarily by synthesis within thebulblets with possible contributions via translocation fromthe mother bulb scales, roots and shoots. Gibberellin A₁₃ wastentatively identified by gas-liquid chromatography but theother components of the bulbiet extracts remained unidentified.     

CHANGES IN THE AMOUNT OF GIBBERELLIN-LIKE SUBSTANCES IN THE SEEDLING OF PHARBITIS NIL WITH SPECIAL REFERENCE TO EXPANSION OF COTYLEDON

Two gibberellin-like substances were separated chromatographicallyfrom the ethanol-extract of germinating cotyledons of Pharbitisnil. In the dark-grown seedling, these substances in the cotyledonincreased in amount gradually with the slow increase in freshweight and area of the tissue, and attained, 4 days after sowing,a maximum level of 0.03 µg gibberellin A₃ equivalent pera couple of cotyledon; in the light-grown seedling, they increasedrapidly with the rapid increase in the fresh weight and area,and attained a maximum content of 0.07 µg gibberellinA₃ equivalent per cotyledon on the 6th day. When the etiolatedseedling was exposed to light, the amount of the gibberellin-likesubstances in the cotyledon increased in parallel with openingof the hypccotyl hook and expansion of the cotyledon. The openingof the hook and increase of the substances occurred in red lightas well as in far-red. Relatively small amounts of the gibberellin-like substanceswere also contained in the germinating hypocotyl and root. The growth of cotyledonary disks was promoted by gibberellinA₃ and the gibberellin-like substances from the expanding cotyledon. The dwarf rice seedling and dwarfs 1, 3 and 5 of maize respondedto the gibberellin-like substances, but dwarf Pharbitis didnot. (Received August 20, 1963; )     

Gibberellin and Inhibitor Content during Iris Bulb Development

Concentrations of gibberellin-like substances and inhibitors were determined in different organs from Iris hollandica cv. Prof. Blaauw during cold treatment and during growth in the greenhouse. Barley-aleurone, dwarf pea and Avena coleoptile straight growth bioassays were used to determine the activity. Developing bulblets contained the highest level of gibberellin activity followed by that in floral organs. During the development of the plants quantitative changes occurred both in the non-acidic, free, and bound pools of gibberellins. Two kinds of inhibitors, “acidic” and “non-acidic”, have been found in the extracts. Most of the non-acidic type were present in the roots. The acidic inhibitors were present in scales and sheath-leaves and disappeared with cold treatment.     

Variations in Endogenous Gibberellins in Developing Bean Seeds II. Changes Induced in Acidic and Neutral Fractions by GA(1)

Immature (8-mm), medium mature (11-mm), and mature green (16- and 17-mm) bean seeds (Phaseolus vulgaris L. cv. Kentucky Wonder and Bountiful) were incubated in gibberellin A₁ solutions for 24 hours at 20°. Extracts from the seeds were separated into nonacidic, acidic ethyl acetate, and acidic butanol fractions. These were chromatographed. The eluates of the chromatograms were tested on Progress No. 9 dwarf peas grown under red light. The level of neutral gibberellin-like substances remained unchanged in immature seed, but they increased markedly in mature green seeds. Coincident with increased levels of the neutral substances, there were significant decreases in acidic ethyl acetate-soluble gibberellin-like substances, including applied GA₁, and in 1 acidic butanol-soluble gibberellin-like substance. Seed incubation in GA₁ brought about increased activity of substance B-II in immature and medium mature seeds. The level of butanol-soluble gibberellin-like substance B-I in seeds of any size was not affected by incubation in GA₁. Considering the marked increases in activity induced in the neutral fraction and the decreases in activity of certain eluates from the chromatograms of the acidic fractions, it was concluded that the neutral fraction may serve as a reserve form of gibberellins in the dry seed. The acidic ethyl acetate substances and substance B-II may be required for normal development of the bean seed.     

Ein Beitrag zur Analyse der Gibberelline in Samen von Pisum sativum L.

Summary Unripe seeds of Pisum sativum were analyses for gibberellins and gibberellin-like substances. Forty kg of seeds were extracted with methanol, the methanol was evaporated and the residue distributed between water and ethylacetate, and between water and butanol at different pH-values. The acidic ethylacetate and the butanol fractions were separated by thin layer chromatography. In the acidic ethylacetate fraction a new substance tentatively named gibberellin A_x was isolated by preparative thin layer chromatography. In different solvent mixtures this substance migrates closely behind gibberellin A₃. It is biological active in a dwarf pea and in dwarf maize mutant bioassays. IR and fluorescense spectra point to a gibban structure. Furthermore gibberellin A₅ was found to be the main gibberellin in pea seeds. It was identified by its R_f-values and the R_f-values of its methylester in different solvent systems as well as by its characteristic relative biological activity in three biological test systems. Most probably gibberellin A₆ is also present in the pea seeds, but because the amounts detected were small, no clear evidence can be presented.In the butanol phase two gibberellin-like substances could be separated by thin layer chromatography.

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Mit Hilfe der Deutschen Forschungsgemeinschaft.     

Extractable and Diffusible Gibberellins From Light- and Dark-grown Pea Seedlings

Gibberellins were obtained from light- and dark-grown peas by solvent extraction and agar diffusion. Both A₅- and A₁-like gibberellins were obtained by extraction; however, by diffusion only the A₁-like gibberellin was found. There was no significant quantitative difference in the levels of diffusible or extractable gibberellin obtained from light- and dark-grown tall and dwarf peas. Several possible explanations for the discrepancy between diffusible and extractable gibberellin were investigated. Of these, only I was supported by experimental evidence, namely, that GA₅ can be converted to GA₁.     

Growth Hormone Changes in Chrysanthemum morifolium: Effects of Environmental Factors Controlling Flowering

Simultaneous quantitative analyses have been made of the endogenouslevels of auxin- and gibberellin like substances, growth inhibitors,and auxin-oxidizing enzyme activity in the cold-requiring Chrysanthemummorifolium cv. Sunbeam subjected to different daylength, lightintensity and temperature regimes known to affect flowering.While little hormone or enzyme activity was found in extractsfrom unvernalized plants, a striking rise in auxin-oxidizingenzyme activity occurred rapidly after the end of cold treatment.Increased auxin activity was also recorded shortly after vernalization.At 28 °C both enzyme and auxin activity declined over aperiod of 3–4 weeks; at 20 °C this response was delayed.Gibberellin activity at 28 °C rose steeply about 2 weeksfrom vernalization and declined several weeks later; at 20 °Ca similar response was less marked. Low light intensity treatment,which may have increased endogenous auxin levels, or exogenousauxin application reduced gibberellin-like substance levelsand cause d devernalization.Phosphon D treatment also loweredgibberellin levels and prevented flowering. An extract fromvernalized plants containing gibberellin-like substances intensifiedthe flowering of partially vernalized test plants. Persistenceof high auxin activity in vernalized plants on long days wasassociated with failure to form normal flower buds. Stem elongationrates correlated in general with levels of endogenous auxin-and gibberellin-like substances. Significant amounts of an abscisin-likeinhibitor were found in extracts of flower buds. The mechanismof natural devernalization is discussed in relation to theseobservations.     

CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

Gibberellins in Light-Grown Shoots of Pisum sativum L. and the Influence of Reproductive Development

The presence of GA₉, GA₁₉ and GA₂₀ was demonstrated by gas chromatography/massspectrometry (GC/MS) and the presence of GA₄₄ strongly indicatedby GC/MS in selected ion monitoring mode (GC/SIM) in extractsof shoots of light-grown tall peas (Pisum sativum L.). Usingthe rice seedling bioassay with cv. Tan-ginbozu, the levelsof gibberellins in pea shoots were monitored from early shootgrowth through to apical senescence in a tall pea line. Levelsof activity corresponding to GA₂₀, GA₁₉ and GA₄₄ remained relativelystable in the shoot despite reproductive development and apicalsenescence. The level of GA₁-like activity increased to a maximumwhen the leaves had between 7 and 9 leaves expanded and decreasedonly with apical senescence. The na gene which blocks the productionof biologically active gibberellins in shoots but not in developingseed, was also operative in pod walls, with na pods containinglittle or no significant gibberellin-like activity when comparedto na pods at contact. This occurred despite the presence ofrelatively high levels of gibberellins in developing seed atthe same time. The results suggest that there is little or nosignificant leakage of biologically active gibberellins fromdeveloping seed to pods or shoots. Extracts of pods of tallpeas with Na contained low levels of gibberellin-like activitybut like developing seed, contained little or no significantGA₁-like activity despite the presence of significant GA₁-likeactivity in shoot extracts of tall peas. (Received March 11, 1986; Accepted May 27, 1986)     

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; )     

The Effect of the Plant Growth Retardants Amo 1618 and CCC on Gibberellin Production in Fusarium monili-forme: Light Stimulated Biosynthesis of Gibberellins

Through the use of a single gene dwarf mutant of Zea mays L., dwarf-1, the interaction of growth retardants with gibberellin biosynthesis was studied in Fusarium monitiforme. It was demonstrated that the growth retardants 2-isopropyl-4-dimcthylamine-5-methyphenyl-1-piperidine-cai'boxylate methyl chloride (Amo 1618) and (2-chloroethyl) trimethylammonium chloride (CCC) are more effective inhibitors of gibberellin biosynthesis in cultures maintained under continuous illumination. Light grown cultures produced significantly more biologically active gibberellin-like materials than dark grown cultures. Stock cultures exposed to light also promoted the subsequent biosynthesis of gibberellins in the dark. Chromatographical analysis of the soluble gibberellins extracted from the culture medium revealed that large amounts of chromatographically detectable A₃ and A₇ were produced in light cultures with only A₇ produced in the dark. Light also induced a greater incorporation of acelate-2-¹⁴C into the gibberellins A₇, A₃ and an unidentified gibberellin. Growth returdants occasionally caused a complete disappearance of chromatographically detectable gibberellins in the dark; however, in the light at no concentration tested was it possible to detect the complete disappearance of gibberellin-like material. A₃ was always detectable. Like higher plants, different strains of F. moniliforme exhibit variation which makes them more or less sensitive to the growth retardants. This variation is interpreted to mean that there may be more than one pathway leading to the synthesis of the gibberellins.     

GIBBERELLIN-LIKE SUBSTANCES OCCURRING IN THE SEED OF PHARBITIS NIL CHOIS. AND THEIR CHANGE IN CONTENTS DURING THE SEED DEVELOPMENT

The amount of gibberellin-like substances in the seed of Pharbitisnil increased in parallel with the growth of the seed, and attained20 days after anthesis to a maximum of 0.115 µg gibberellinA₃ eqivalent per seed, when the seed reached its maximum freshweight or four-fifths of its final dry weight. At this maximumlevel, 0.03 µg and 0.1 µg gibberellin A₃ equivalentswere localized in the embryo and in the "endosperm", respectively.Three gibberellin-like substances (Factors I, II and III) wereseparated upon paper chromatography. In view of the changesin amount of the factors with respect to the seed maturation,these factors, especially Factor II, in the "endosperm" andembryo were assumed to participate in the initial or the mainpart of growth of the embryo. Dwarf rice seedling and maizedwarfs 1, 3 and 5 responded to the three factors nearly in thesame way. Pharbitis dwarf, however, responded only to FactorI, but not to Factors II and III. (Received February 21, 1963; )     

Internode length in Pisum. III The effect and interaction of the Na/na and Le/le gene differences on endogenous gibberellin-like substances

In the garden pea ( Pisum sativum L.), shoots of the extremely short plants with the mutant na (phenotype nana) are found by bioassay to contain undetectable levels of gibberellin-like substances. This is confirmed by the use of near isogenic lines differing at the Na locus. Thus, mutant na appears to block a step early in the pathway of gibberellin synthesis. It is suggested that the polar gibberellin-like substance found in the apical portion of shoots of tall ( Le ) but not dwarf ( le ) peas could be GA₁. Extracts of shoots of na Le peas treated with GA₂₀ (the major active gibberellin in dwarf peas) possess a large amount of GA₁-like activity whereas extracts of shoots of na le peas treated with GA₂₀ possess a much reduced amount. Thus, gene Le may allow the conversion of a less active gibberellin (GA₂₀) into one more active in stimulating elongation in the pea (the GA₁-like compound). In contrast to their influence in the shoot, the na and Le genes do not appear to be operative in controlling the gibberellin content of developing seed, indicating that organ specific gibberellin biosynthesis and metabolism occur in peas.     

Reduction of the Gibberellin Content of Pharbitis Seeds by CCC and After-Effects in the Progeny

Plants of Pharbitis nil were treated with the growth retardant (2-chloroethyl) trimethylammonium chloride (CCC) shortly before and after anthesis. Fresh and dry weight of immature seeds were not affected by the CCC treatment.

The level of gibberellin-like activity in Pharbitis seeds as compared to control seeds was strongly reduced by CCC application. The progenies of the treated plants also had a much reduced GA content in the seedling stage. These results are interpreted to indicate that CCC blocks gibberellin biosynthesis in higher plants, as it does in the fungus Fusarium.

CCC applied via the roots accumulated in the immature seeds and was carried over to the following generation. Consequently, growth of CCC progenies was dwarfed and flower formation inhibited. Both phenomena were overcome by application of gibberellin A₃.

Three gibberellin-like substances (called fractions I, II, and III) were present in Pharbitis seeds and could be separated by thin-layer chromatography. All 3 fractions were also present in seeds treated with CCC. Fractions II and III were present in much higher quantities than fraction I. Both fractions II and III promoted growth of d5 corn but only fraction II was active in dwarf peas grown under red light.

     

Comparison of gibberellin-like substances from cotyledons and phloem exudate collected from cotyledons of Pharbitis nil in relation to photoperiodic flowering

Gibberellin (GA)-like substances were analyzed in extracts from cotyledons and phloem exudate collected from cotyledons in photoinduced and vegetative seedlings of the short-day plant Pharbitis nil Chois. var. Violet, using high performance liquid chromatography (HPLC) and the dwarf rice bioassay, to see whether any specific GA-like substances were transported from the photoinduced cotyledons via phloem. Cotyledon extracts exhibited five peaks of free GA-like activity in HPLC, whereas only one or two active peaks were detected in phloem exudate extracts. The level of free GA-like activity was considerably lower in phloem exudate than in the cotyledons. In five out of six analyses of cotyledons and phloem exudate, there were substantially higher levels of free GA-like substances in photoinduced plants. Conjugated GA-like substances were present in much higher levels than free GA-like substances in the cotyledon extracts but the levels were not influenced by daylength. In phloem exudate extracts there was no conjugated GA-like substances. The free GA-like substances that are transported via phloem cochromatographed with GA_5/20 and GA₁₉ on HPLC. These were significantly higher in photoinduced plants and thus could have some influence on the photoperiodically-induced flowering in P. nil.     

Evidence for the Translocation of Gibberellin A3 and Gibberellin-Like Substances in Grafts between Normal, Dwarf1 and Dwarf5 Seedlings of Zea mays L.

Approach grafts were made between the cut surfaces of mesocotylsfrom normal and dwarf seedlings of Zea mays L. (maize). Thedwarfs were the non-allelic single gene gibberellin mutants,dwarf₁ and dwarf₅. The graft combinations were normal-normal,normal-dwarf₁, normal-dwarf₅, dwarfi-dwarf₁, dwarf₅-dwarf₅,and dwarf₁-dwarf₅. The grafts were used to demonstrate the movementof gibberellin-like substances across the union. GA₃, addedto one member of the graft, resulted in leaf-sheath elongationto both members. Gibberellin-like elongation was observed inthe dwarf₁ and dwarf₅ members of normal-dwarf₁ and normal-dwarf₅grafts. Also, in some cases, elongation was observed in thedwarf₁ and dwarf₅ members of dwarf₁-dwarf₅ grafts, especiallyin the dwarf₅ members. The results are discussed in terms of:the gibberellin biosynthetic pathway in maize, the control ofthis pathway by the d₅ and d₁ mutant genes, and the movementof gibberellin-like substances in the plant. Anatomical observationof grafted seedlings after the resonse was observed shows thatcallus cells developed and bridged across the interface of theautografts. There was also evidence for vascular differentiationacross the graft union. ¹ Dedicated to late Prof. J. Ashida. (Received December 17, 1982; Accepted February 9, 1983)     

The growth promoting effect of red light on internode elongation of Progress seedlings

Red light increased elongation of the apical 10 mm of epicotylsexcised from 7-day-old dark grown Progress seedlings. Removalof the basal portion of the plant appears to render the tissueinsensitive to the inhibitory influences of light. Additionof gibberellins A₁, A₃ or A₅ further increased elongation. Thered light growth response was independent of the gibberellinresponse; therefore, it was considered to be unrelated to anincrease in gibberellin biosynthesis. A study of the time course of growth in the presence of thegibberellins revealed that a 6–8 hr lag period was requiredbefore A₁ and A₅ became effective, while no lag period was associatedwith A₃. It was suggested that A₁ and A₅ were converted to anA₃-like gibberellin. (Received July 29, 1972; )     

Levels of gibberellin-like substances and their possible transport in developing dwarf and normal cultivars of pea (Pisum sativum L.)

The level of gibberellin-like substances was determined in the cotyledons and axis of developing seedlings of dwarf (Little Marvel) and normal (Tall Telephone) cultivars of pea (Pisum sativum L.). The effect of cotyledon removal with GA₃ application on growth was also examined. Greater levels of gibberellin-like substances were observed in the cotyledons of the normal cultivar than the dwarf. This was particularly evident in the cotyledons during the early stages of seedling growth. Subsequently there was a decline in GA levels in the cotyledons. This was coincidental with a rise in GA content in the axis with markedly greater levels in the normal than the dwarf cultivar. Decotyledonated dwarf and normal plants supplied with GA were much taller than the decotyledonated controls. This observation along with those of the gibberellin levels in the cotyledons and axis, provided circumstantial evidence that there may be translocation of gibberellins from the cotyledons to the axis.     

The Relationship of Endogenous Gibberellins to Light-regulated Stem Elongation Rates in Dwarf and Normal Cultivars of Pisum sativum L.

Dwarf cultivar Progress No. 9 and normal cultivar Alaska ofPisum sativum L. were grown under conditions of darkness orred light. Red light decreased the stem elongation rate of bothcultivars. Gibberellins present during the linear phase of stemelongation were isolated and the two main components were tentativelyidentified by gas chromato-chromatography and mass spectrometryas Gibberellin A₁ and A₅. Both gibberellins varied quantitativelywithin and between cultivar and treatment groups, and the amountspresent were inversely related to stem elongation rate. Althoughthe stem elongation rate of plants grown under red light wasrepressed, endogenous gibberellin was not limiting and was asmuch as two-fold higher in red light-grown plants than in dark-grownplants. The levels of endogenous gibberellin in dawrf plantsindicated that the genetic growth limitation was not due toa gibberellin deficiency. (Received May 26, 1981; Accepted January 28, 1982)     

Über den Gibberellingehalt von Zwerg- und Normalerbsen im Rotlicht und die Wirkung von Chlorcholinchlorid auf das Wachstum der Erbsen

Zusammenfassung Keimlinge von Zwerg- und Normalerbsen wurden auf ihren Gibberellingehalt getestet. Die Normalerbsen enthalten etwa achtmal soviel wie die Zwerge von der Gibberellinfraktion I nach Kende und Lang, die auf Normalerbsen stark, auf Zwerge kaum wirkt.Beide Rassen wurden mit Chlorcholinchlorid, einem Hemmstoff der Gibberellinsynthese bei Fusarium, behandelt. Im Rotlicht werden die normalen Pflanzen verzwergt, doch im Dunkeln hat die Substanz nur einen geringen Effekt. Die Dunkelhemmung ist so groß wie die Hemmung der Zwerge in Rotlicht. Eine mögliche Erklärung für die Unwirksamkeit von CCC im Dunkeln ist, daß die Erbsen im Dunkeln zum Wachstum kein oder nur wenig Gibberellin benötigen.

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The content of gibberellin-like substances in dwarf and normal peas growing in red light, and the effect of chlorocholinchloride on growth of peas

Summary Dwarf and normal pea seedlings were extracted and their gibberellinlike substances bioassayed. The normal peas yield about 8 times as much as the dwarfs of gibberellin-fraction I after Kende and Lang, which is highly effective on normal peas but nearly without effect on dwarfs.Both cultivars were treated with the inhibitor of gibberellin-synthesis in the fungus Fusarium, chlorocholinchloride (CCC). In red light the normal plants are dwarfed, but in darkness this substance has only a weak inhibiting effect on growth. The inhibition in darkness is of the same magnitude as the effect of CCC on dwarfs growing in red light. A possible explanation for the ineffectiveness of CCC in darkness is that peas need there no gibberellin at all or only very minute amounts for growth.