Establishment of photoperiodic sensitivity by benzyladenine and a brief red irradiation in dark grown seedlings of Pharbitis nil Chois.

Flowering of etiolated seedlings of Pharbitis nil resulted followinga single, brief red irradiation prior to an inductive dark period.Following this irradiation benzyladenine sprayed on the seedlingsenhanced flowering dramatically and this effect was maximalfor concentrations between 44 and 120µM. In the presenceof benzyladenine a brief (4 to 10 sec) low energy red irradiation(2.6 Wm^–2) resulted in flowering and repeated far-redphotoreversal of this red promotion provided clear evidenceof the sole involvement of phytochrome. However, after suchbrief irradiations the critical dark period for flowering waslonger than is normally found in seedlings grown in light whichindicated that additional photoresponses might be importantin natural conditions. An examination of seedling photosynthesisand assimilate transport indicated that the benzyladenine effecton flowering may relate to its promotion of assimilate and floralstimulus transport to the shoot apex. ¹ Present address: Faculty of Agriculture, Mie University, TsuCity, Mie Prefecture, Japan. (Received August 21, 1978; )     

Flower-Inducing Activity of Phloem Exudates from Pharbitis cotyledons Exposed to Various Photoperiods

The phloem exudate prepared from the cotyledons of Pharbitisseedlings that had been exposed to a single dark period (oflonger than 10 h) induced flowering in cultured apices excisedfrom non-induced seedlings. The flower-inducing activity ofthe exudate increased as the seedlings were exposed to longerperiods of darkness. The highest activity was associated withthe exudate taken from cotyledons exposed to a single 16-h darkperiod. The activity of the exudate taken from cotyledons exposedto an inductive dark period was clearly reduced by interruptionof the dark period. The addition of exudate taken from threecotyledons to 10 ml of medium resulted in the highest flower-inducingactivity. About 50% of cultured apex explants formed floralbuds, even when the concentration of the exudate was reducedto 0.1 cotyledon equivalents per 10 ml of medium. The flower-inducingactivity of the exudate appeared to be heat-stable. (Received December 13, 1991; )     

The Negative Response of Photoperiodic Floral Induction in Chenopodium rubrum L. to Preceding Growth

In Chenopodium rubrum there exists a correlation between theage of the seedlings and the effectiveness of photoperiodicinduction. The younger the plants the more effective was photoperiodictreatment. In three-day-old seedlings one short day was sufficientto promote incomplete flowering, while two short days broughtabout 100 per cent flowering. With six-, eight-, and ten-day-oldplants exposed to two or three short days quantitative differenceswere observed in the earliness of flowering and the percentageof flowering plants. The effects of continuous light and ofshort days with a light break preceding the inductive treatmentwere compared. The results obtained indicate that the inhibitoryeffect of plant age cannot be attributed solely to the appearanceof inhibitors under continuous light but changes of growth patternin plants of different age should also be taken into consideration. The inhibition of RNA synthesis in shoot apices brought aboutby 6-azauridine resulted also in a flowering stimulation, providedthat the inhibitor was applied one or two days prior to inductionand the inductive process itself remained undisturbed. Thisstimulation was accompanied by inhibition of vegetative growthand by a decrease of RNA concentration in the cytoplasm as estimatedcytophotometrically. The competition between growth of vegetative organs and floraldifferentiation affects the response to inductive treatment.The suppression of growth can result in enhancement of flowering.     

Site of Perception of the Far-Red Inhibition of Flowering in Pharbitis nil Choisy

The site of perception of the inhibition of flowering by far-redlight (729 nm) was investigated in the short-day plant Pharbitisnil, using a specially designed optical fibre apparatus withinterference filters. The cotyledon was found to be the primarysite of perception and no inhibition was obtained when the apexalone was irradiated. However, when the apex and cotyledon weresimultaneously irradiated there was a reduction in the numberof terminal flower buds compared with the cotyledon alone. Forwhole plants, far-red light centred at 729 nm (10 nm bandwidth)was considerably more inhibitory than at 719 nm. ¹Present address: Department of Biology, Guru Nanak Dev University,Amritsar—143005, Panjab, India. (Received April 28, 1986; Accepted June 23, 1986)     

Studies on the Photoreceptors for the Promotion and Inhibition of Flowering in Dark-Grown Seedlings of Pharbitis nil Choisy

Three-day-old etiolated seedlings of Pharbitis nil were exposedto red light for 10 min and sprayed with N⁶-benzyladenine beforetransfer to a 48-h inductive dark period, after which they weregrown under continuous white light. A second red irradiationpromoted flowering when given at the 5 and 24th hour of theinductive dark period but inhibited flowering at the 10 and15th hour. Far-red light inhibited flowering when given at anytime during the first 24 h of the dark period. Red/far-red reversibilitywas clearly observed at the 0, 5, 10 and 24th hour, but notat the 15th hour when both red and far-red lights completelyinhibited flowering. The action spectrum for the inhibition of flowering at the 15thhour of the inductive dark period had a sharply defined peakat 660 nm and closely resembled the absorption spectrum of theP_R form of phytochrome. The photoreceptors involved in thesephotoreactions are discussed. (Received June 10, 1983; Accepted July 6, 1983)     

Phloem Exudate of Perilla crispa and its Effects on Flowering of P. crispa Shoot Explants

The aim was to develop an assay for the flowering stimulus ofa photoperiodically-sensitive plant. Phloem exudate solutionswere obtained from photoperiodically induced and non-inducedleaves of Perilla crispa (Thunb.) Tanaka, following treatmentof excised leaves with solutions of EDTA or phytic acid. Theamounts of exudate obtained were estimated polarimetrically,and the conditions for obtaining maximum exudate yields weredetermined. Shoot explants from non-induced P. crispa plantswere grown on a nutrient medium. Under short days the explantsreached anthesis after c. 35 d. In continuous light a smallproportion of the explants showed signs of flowering after 100d. The effects of test substances and phloem exudate on theflowering of explants grown in continuous light was investigated.(?)-ABA (4.0 µM), sucrose (14.6 mM) and phloem exudatefrom both induced and non-induced leaves caused some promotionof flowering. In three experiments, phloem exudate from inducedleaves enhanced flowering to a greater extent than exudate fromnon-induced leaves; in other experiments the effects of thetwo types of exudate were similar. There was no evidence thatABA or sucrose in the phloem exudate caused flowering. Concentrationsof phloem exudate above 2.0 g I^–1 were phytotoxic to theexplants. Key words: Chelating agents, Flowering, Perilla crispa, Phloem exudate, Phytic acid, Shoot culture     

[14C]-Assimilate partitioning in photoperiodically induced seedlings of Pharbitis nil. The effect of benzyladenine

Partitioning of [¹⁴C]-labeled assimilates was studied in relation to photoperiodic floral induction and evocation in one-week-old Pharbitis nil Choisy cv. 'Violet' seedlings. In plants kept under 16 h photoperiods, one 15 h night induced 100% axillary flowering whereas a 24 h night induced both terminal and axillary flowering. A 15 min night break of red light given 8 h after the beginning of the dark period inhibited flowering. Total [¹⁴C]-assimilate distribution among major sinks (plumules + epicotyl and roots + hypocotyl) from a single source cotyledon was unchanged by one inductive night; however, import of [¹⁴C]-assimilates into shoot apices was increased in induced plants compared to vegegative controls. This increase was several-fold in plants subjected to a 24 h night. N⁶-Benzyladenine (BA) application to cotyledons or plumules under non-saturating night lengths increased the number of floral buds per plant without affecting the position of the first floral bud (i.e. the speed of induction). The same treatment caused increased label accumulation in induced apices, while it only slightly affected non-induced ones. The mode of action of BA on flowering through growth stimulation and resulting assimilate mobilization is discussed.     

Indirect Action of Benzyladenine and Other Chemicals on Flowering of Pharbitis nil Chois: Action by Interference with Assimilate Translocation from Induced Cotyledons

Benzyladenine (BA) brushed on the cotyledons of 4-day-old seedlings of Pharbitis nil Chois. markedly stimulates flowering. Greates response is obtained for concentrations between 44 and 440 micromolar. The action of BA is on processes in the cotyledon as shown by the response to its site of application, to the dosage applied and to the requirement for its application prior to the dark period. There was little or no effect of BA treatment on either the time measurement processes of photoperiodic induction or on the generation of floral stimulus. Transport of photosynthetic assimilate from the cotyledons to the shoot apex was altered.     

Effects of Indol-3yl acetic Acid on Floral Induction and Apical Differentiation in Chenopodium rubrum L.

Indol-3yl acetic acid (10^–4M) was applied to the plumulesof Chenopodium rubrum. Effects on the anatomical structure andthe growth pattern in the apical meristem, as well as DNA synthesisand nucleolus size were investigated. When auxin is applied before or during photoperiodic inductionit inhibits DNA synthesis and meristematic activity. The axillarymeristem (i.e. a group of cells in the axils of the leaf primordia)is most affected. A similar inhibition of the axillary meristemwas also observed in non-induced control plants grown in continuouslight. Auxin applied simultaneously with photoperiodic inductioncounteracts the reduction of apical dominance in the apex andthus inhibits the onset of floral differentiation. Auxin appliedfollowing induction inhibits the previously-formed buds andmakes possible a more complete development of the apical flower. The dual effect of IAA on flowering, inhibitory and stimulatory,manifests itself as a growth response at different stages ofthe changing shoot apex.     

Stimulation of the Flowering of Pharbitis nil Chois. by Gibberellin A3 : Time Dependent Action at the Apex

Gibberellin A₃ (GA₃) stimulated flowering when it was appliedto the shoot apex of seedlings of Pharbitis nil, dwarf strainKidachi; but, not when it was applied to the cotyledons. GA₃applied to the plumule before or shortly after the start ofan inductive dark period promoted both flowering and shoot elongation;but, the later the time of application during the dark periodless the promotion of flowering, although marked promotion ofshoot elongation always took place. The variation with time in the response of flowering to GA₃indicates that early floral processes at the apex are stimulatedby GA₃, but that subsequent processes are insensitive to it.The early processes of floral stimulus produced by a 16 hr inductivedark period probably are completed within 20 hr at 28°Cafter the end of the dark period. At low temperatures, suchas 15 and 20°C, early floral processes continued for morethan 40 hr. When cotyledons were removed at various times, the export ofthe floral stimulus to the shoot apex was apparent within hoursof the generation of the floral stimulus in the cotyledons,which started with the passage of the critical 9-hr dark period. (Received February 18, 1981; Accepted March 24, 1981)     

Inhibition of Flower Induction by Some Inhibitors of Protease in the Short-day Plant Lemna paucicostata 6746 and the Long-day Plant Lemna gibba G3

Four inhibitors of proteases, namely, bestatin, diisopropylfluorophosphate, elastatinal and p-toluenesulfonyl-L-lysinechloromethyl ketone hydrochloride, were examined for their effectson flowering of a short-day plant Lemna paucicostata 6746 anda long-day plant Lemna gibba G3. Each of the inhibitors greatlyinhibited the flowering of Lemna paucicostata 6746 that is normallyinduced by nitrogen deficiency. Bestatin or elastatinal givenonly during the first half of the culture period inhibited theflowering more clearly than when each was given during the latterhalf, suggesting that they inhibited the inductive process(es)involved in flowering rather than development of flower buds.Bestatin or elastatinal greatly inhibited the flowering of Lemnapaucicostata 6746 induced by photoperiodic stimulus, ferricyanideand continuous far-red light. Simultaneous application of thesetwo inhibitors was more effective in the inhibition of photoperiodicallyinduced and ferricyanide-induced flowering than was each inhibitoralone. They also completely inhibited the photoperiodic floweringof Lemna gibba G3. These results suggest that the inductionor activation of some proteases, probably followed by the degradationof some protein(s), is necessary for the induction of floweringin both these plants. (Received November 21, 1989; Accepted February 19, 1990)     

Effects of Abscisic Acid on the Growth Pattern of the Shoot Apical Meristem and on Flowering in Chenopodium rubrum L.

Abscisic acid (ABA) at 1 x 10^–4 M or 3 x 10^–4 Mwas applied to the apical buds of Chenopodium rubrum plantsexposed to different photoperiodic treatments and showing differentpatterns of floral differentiation. Stimulation of growth inwidth of the apical meristem of the shoot and/or inhibitionof growth in length was obtained under all photoperiodic treatments.This change of growth pattern was followed by different effectson flowering. In non-induced plants grown under continuous light ABA stimulatedpericlinal divisions in the peripheral zone and the initiationof leaves as well as the growth in width of bud primordia. Inplants induced by two short days reduced growth of the meristemcoincided with ABA application. Longitudinal growth of the meristemwas inhibited in this case and only a temporary stimulationof inflorescence formation took place. In plants induced ata very early stage, ABA exerted a strong inhibitory effect onflowering. A permanent and reproducible stimulatory effect onflowering was obtained in plants induced by three sub-criticalphotoperiodic cycles if ABA was applied to apices released fromapical dominance. In this case formation of lateral organs andinternodes was promoted by ABA and was followed by stimulatedinflorescence formation. Gibberellic acid (GA2) at 1x 10^–4M or 3 x 10^–4 M brought about a similar effect on floweringas ABA, although the primary growth effect was different, i.e.GA₂ stimulated longitudinal growth. The effects of ABA and GA₂ on floral differentiation have beencompared with earlier results obtained from auxin and kinetinapplications. These growth hormones are believed to regulateflowering by changing cellular growth within the shoot apex.Depending on the actual state of the meristem identical growthresponses may result in different patterns of organogenesisand even in opposite effects on flowering. Shoot apex, flowering, photoperiodic induction, abscisic acid, gibberellic acid, Chenopodium rubrum L.     

Changes in cotyledon mRNA during floral induction of Pharbitis nil CV. Violet

Floral induction in seedlings of Pharbitis nil Choisy cv. Violet, with one cotyledon removed, was manipulated by applying various photoperiodic treatments to the remaining cotyledon. Populations of polyadenylated RNA from treated cotyledons were examined to identify messages specifically involved in floral induction. The RNA was translated in vitro using a wheat-germ system, and the resulting translation products were analysed by two-dimensional polyacrylamide gel electrophoresis. Substantial qualitative and quantitative differences were found between mRNA from cotyledons of seedlings kept in continuous light (non-induced) and of seedlings given a 16-h dark period (induced). In contrast, inhibition of flowering with a night-break resulted only in one detectable, quantitative difference in mRNA.Abbreviations CL continuous light - kDa kilodalton - NB 16 h darkness+10 min red-light break, 8 h into the dark period - poly(A)⁺ RNA polyadenylated RNA (isolated by binding to a cellulose oligodeoxythymidine affinity column) - SD short day (16 h dark) - SDP short-day plant - SDS sodium dodecyl sulfate     

Participation of polyamines in the flowering of the short-day plant Pharbitis nil

The effect of the exogenous application of polyamines on the flowering induction of the short-day plant Pharbtis nil was investigated. Putrescine, spermidine and spermine applied on the cotyledons of 4-day seedlings had no significant effect on the flowering of this plant under conditions of full induction caused by a 16-hour-long inductive night. Under the conditions of partial induction caused by a 13-hour-long subinductive night, polyamines inhibit or stimulate flowering, depending on the time of application. Also, inhibitors of the biosynthesis of polyamines influenced the flowering process. Analysis of endogenous polyamines revealed significant fluctuations in their content in cotyledons during an inductive night, as well as under continuous light conditions. Particularly large changes occurred in spermidine and spermine levels. The putrescine level in induced seedlings was lower than in non-induced ones. However, induced seedlings contained a higher level of spermine and spermidine. The highest spermidine and spermine levels were observed at the 8th h of the night, although the total concentration of spermine during photoinduction was always 2–3 times lower than that of spermidine. A break in the inductive night, leading to a complete inhibition of flowering, had caused significant changes in the polyamine level by the end of the night. The results suggest that the flowering induction of Pharbitis nil took place at a low putrescine level and increased spermidine and spermine levels.     

Inhibition of photoperiodic floral induction in Pharbitis nil by ethylene

Application of ethylene at 100 ppm or higher completely inhibitedflowering in Pharbitis nil when made during an inductive darkperiod. Exposing plants to ethylene before or after the inductivedark period produced only slight or almost no inhibition. Ethylenewas effective when it was applied only to a cotyledon, but wasineffective when applied only to a receptor bud. Ethylene hadno effect on translocation of the floral stimulus. Ethylene-treatedcotyledon did not transfer any flower inhibiting entity. Thus,ethylene is considered to inhibit the induction process(es)in cotyledons. Except for an initial temporary cotyledon epinasty, ethylenetreatment had no effect on the subsequent growth and vigor ofplants. This temporary cotyledon epinasty disappeared withinthe next 24 hr. (Received May 4, 1972; )     

Light requirement,phytochrome and photoperiodic induction of flowering of Pharbitis nil Chois

For dark-grown seedlings of Pharbitis nil capacity to flower in response to a single inductive dark period was established by 24 h white, far-red (FR) or ruby-red (BCJ) light and by a skeleton photoperiod of 10 min red (R)-24 h dark-10 min R. FR alone was ineffective without a brief terminal (R) irradiation, confirming that the form of phytochrome immediately prior to darkness is a crucial factor for flowering in Pharbitis. The magnitude of the flowering response was significantly greater after 24 h FR or white light (WL) (at 18° C and 27° C) than after two brief skeleton R irradiations, but the increased flowering response was not attributable to photosynthetic CO₂ uptake because this could not be detected in seedlings exposed to 24 h WL at 18° C. Photophosphorylation could have contributed to the increased flowering response as photosystem I fluorescence was detectable in plants exposed to FR, BCJ, or WL, but there were large differences between flowering response and photosystem I capacity as indicated by fluorescence. We conclude that phytochrome plays a major role in photoresponses regulating flowering. There was no simple correlation between developmental changes, such as cotyledon expansion and chlorophyll formation during the 24-h irradiation period, and the capacity to flower in response to a following inductive dark period. Changes in plastid ultrastructure were considerable in light from fluorescent lamps and there was complete breakdown of the prolamellar body with or without lamellar stacking at 27 or 18° C, respectively, but plastid reorganization was minimal in FR-irradiated seedlings.Abbreviations BCJ irradiation from photographic ruby-red lamps - FR far-red light - P_fr far-red-absorbing from of phytochrome - P total phytochrome content - R red light - WL white light from fluorescent lamps     

Influence of Chloride and Transpiration on Net15NO3-Uptake Rate byCitrus Roots

Three-month-old Carrizo citrange (hybrid of Citrus sinensisL. OsbeckxPoncirus trifoliata Blanco) seedlings were grown incontrolled environment chambers in pots of fine sand. Plantswere irrigated with either non-saline or saline solutions overa 3-week period. After these treatments, plants were transferredto vessels containing a 5 m M¹⁵NO₃K (96% atom excess¹⁵N) solution,and transpiration as well as concentration of¹⁵N and Cl^-in roots,stem and leaves were measured after 24 h. Transpiration and¹⁵NO₃^-uptakerates were inhibited after exposure to NaCl and the concentrationof salt pre-treatment determined the intensity of this inhibitoryeffect. To determine the effect of transpiration on NO₃^-absorption,net¹⁵NO₃^-uptake rate was measured in salt stressed and non-stressedplants exposed to different light intensities or relative humiditiesand also in detached roots. Reduction in NO₃^-uptake was moreclosely related to Cl^-antagonism from salt stress than to reducedtranspiration rate. Copyright 1999 Annals of Botany Company Nitrate, absorption, inhibition transport system, salt, light and humidity.     

BLUE LIGHT, PHYTOCHROME AND THE FLOWERING OF LEMNA PERPUSILLA 6746

Lemna perpusilla 6746 was grown on HUTNER'S medium with sucroseunder light schedules combining red, blue and far-red light.As shown earlier, brief red exposures added to a continuousblue schedule inhibit flowering although either schedule alonepermits it; hence red and blue act together in establishinga long day (flower inhibiting) condition. However, the red exposurerequired to inhibit flowering is greater with high intensitythan with low intensity continuous blue, suggesting in additiona blue-red antagonism. Blue light reverses the effects of abrief red exposure closing a blue or far-red main photoperiod,but it also reverses the effects of a brief far-red exposureclosing a red photoperiod. Thus, blue can act either like redor like far-red, depending on the situation. All effects ofblue light on the flowering of L. perpusilla 6746 are consistentwith the notion that it establishes a Pfr level intermediatebetween those established by red and far-red light; the postulationof an additional photoreaction to explain the effects of blueseems unnecessary. ¹Research carried out at Brookhaven National Laboratory underthe auspices of the U. S. Atomic Energy Commission.     

Growth and Development in the Young Tomato: I. THE EFFECT OF TEMPERATURE AND LIGHT INTENSITY ON GROWTH OF THE SHOOT APEX AND LEAF PRIMORDIA

Tomato seedlings were grown at constant temperatures of 25°and 15° C. in a 12-hour day at light intensities of 1,600,800, and 400 f.c. The rate of increase in size of the shootapex and the rates of formation and growth of leaf primordiaduring the vegetative phase were followed by dissecting samplesfrom the time of cotyledon emergence onwards. The rate of enlargement of the shoot apex increased with lightintensity, but apical enlargement was delayed at the highertemperature, the delay being longer the lower the light intensity.The rates of leaf formation and leaf growth increased with bothtemperature and light intensity. Temperature had a larger effecton leaf growth than on leaf formation. More leaves were formedbefore flowering at 25° C. than at 15° C., the increasein leaf number being greater the lower the light intensity. It is suggested that the delay in the enlargement of the apexat high temperature can be explained in terms of competitionfor assimilate, the competitive potential of the expanding leafprimordia exceeding that of the apex at higher temperatures.     

The Growth of the Shoot Apex of Chenopodium rubrum L. Treated with a Florigenic Extract from Flowering Tobacco Plants: Preliminary Anatomical Observations

Anatomical changes in the shoot apex of Chenopodium rubrum L.treated with an extract from flowering tobacco plants and cultivatedin non-inductive conditions are described. They are comparedwith the anatomy of non-treated vegetative apices and with apicesof plants induced with a short day. Treatment with the extractresulted in both activation of cell division in the upper partof the apex and in apex elongation. Acceleration of leaf primordiainitiation and stimulation of branching took place. The effectcorresponds to the sequence of changes in photoperiodically-inducedplants but is more pronounced. Elongation following 10^–4M GA₃ treatment was of a differentnature; there was only a slight stimulation in the upper partof the apex in contrast with a strong stimulation of growthin length in the lower internodes. These preliminary resultssuggest a similarity between apical changes evoked by a stimulusproduced by short days and an exogenously applied floral stimulus.The changes differed from those caused by exogenous phytohormones. Key words: Chenopodium rubrum, florigen, shoot apex