The relationship between red and far-red light on flowering of the long-day plant, Lemna gibba

Lemna gibba, a long-day duckweed, can be induced to flower whenthe 10 hr white photoperiod is extended with red or far-redlight. The 10 hr red photoperiod is also effective in inducingflowering when followed by a far-red extension, but a red extensionis ineffective. When 2 hr of far-red light are given immediately after the 10hr red photoperiod, the following red as well as the far-redextension can induce flowering, indicating that the 2 hr far-redlight plays an important role as a starting factor for induction.This red or far-red extension is effectively replaced by a redbreak given at a proper time in the darkness which follows the2 hr far-red light as the starting factor. The effect of thered break in not cancelled by subsequent exposure to far-red,which synergistically promotes flowering. However, a red break given immediately after a proper periodof far-red extension further promotes flowering. The phase sensitiveto the red break coincides with that sensitive to the red breakgiven in darkness. The effect of the red break is reversed bysubsequent exposure to far-red, contrary to the effect of thered break in darkness. Using these results, relation between red and far-red lighton flowering in L. gibba is discussed. (Received July 17, 1971; )     

Induction of flowering in Lemna gibba G 3 by Fe-EDDHA

Fe-EDDHA (iron salt of ethylenediamine-di-o-hydroxyphenylaceticacid) induced profuse flowering in Lemna gibba G 3 culturedin HUTNER'S medium. The maximum number of flowering plants wasobserved in a medium supplemented with 5 ppm of this chelate. (Received April 20, 1970; )     

Gibberellin-EDDHA interaction in flowering and gibbosity of Lemna gibba G3

The effect of gibberellic acid (GA₃), in the presence of EDDHA,on the flowering and gibbosity of Lemna gibba G3 was studied.At 10 ppm and at higher concentrations of GA₃ the EDDHA-effect,i.e. profuse flowering and conspicuously gibbous fronds, wascompletely nullified. (Received July 15, 1974; )     

Induction of flowering in Lemna gibba G3 under short-day conditions

In the presence of the chelating agent EDDHA, long-day duckweedLemna gibba G3 was induced to flower under a short-day scheduleof 9 hr of light and 15 hr of darkness in a 24-hr cycle. Weconcluded that EDDHA creates effects very similar to those ofsalicylic acid. When EDDHA or salicylic acid was added to thenutrient medium in combination with BA, flowering was inducedeven under conditions of 8 hr of light and 16 hr of darkness.Under a photoperiod of 9 hr, BA markedly enhanced the effectof EDDHA as well as salicylic acid. On the other hand, BA alonewas ineffective as far as flowering was concerned. By quantitativeinteractions, BA seems to complement the modifying effect ofEDDHA or salicylic acid on flowering in this duckweed strain. (Received June 25, 1976; )     

On the rhythm of sensitivity to light interruption in a long-day duckweed, Lemna gibba G 3

1. The rhythm of sensitivity to light interruption in a long-dayduckweed, Lemna gibba G 3, was examined. The rhythm was circadianand was suggested to be under the control of a physiologicalclock. Light given in the trough between the first and secondpeaks reset the rhythm. Five to 7 cycles of non-circadian photoperiodicregimes given entrained the rhythm to external periodicity,and this entrained rhythm persisted even after the plants weretransferred to continuous darkness. 2. It was suggested that the induction period is not determinedby the physiological clock disclosed here, but by a periodicalternation of dark-sensitivity, or by a periodic change inactivity of SS (system sensitive to IPEF, induction period extendingfactor,) as postulated previously. (Received November 28, 1967; )     

Short-day flowering of Lemna gibba G3 induced by salicylic acid

Salicylic acid, probably as a chelating agent of the EDTA-salicylaldoximetype, can eliminate the light requirement during the inductivephase of Lemna gibba G3, and thus is able to induce short-dayflowering of this long-day plant. (Received September 4, 1975; )     

Interaction of naphthol with EDDHA/salicylic acid in flowering and gibbosity of Lemna gibba G3

Flowering and gibbosity in Lemna gibba G3, as enhanced or inducedby optimal concentrations of salicylic acid or EDDHA, was specificallyinhibited by 10 µg/ml and higher concentrations of ß-naphthol.These effects of ß-naphthol are similar to those ofbreakdown products of GA_s. (Received March 20, 1978; )     

Specific interactions in the physiology of flowering and gibbosity of Lemna gibba G3

Fronds of Lemna gibba G3 became conspicuously gibbous when ethrel,an ethylenereleasing compound, was added to the nutrient medium.Maximal gibbosity was obtained at ethrel concentrations of 1µg/ml and higher. Unlike the chelating agent, EDDHA, whichcauses profuse flowering and markedly gibbous fronds under long-dayconditions, ethrel did not affect flowering. In the presenceof an optimal concentration of EDDHA (10 µ/ml), ethreleven significantly inhibited flowering and caused developmentof excessively gibbous fronds. Autoclaved gibberellic acid specifically negated the ethreleffect as it does that of EDDHA. Three decomposition productsof GA₃, allogibberic acid, epiallogibberic acid and gibbericacid, also nullified flowering and gibbosity in the presenceof EDDHA. A fourth decomposition product of GA₃, epigibbericacid, inhibited gibbosity but hardly affected flowering. Salicylic acid was confirmed to affect flowering and gibbosityin L. gibba G3. However, contrary to an earlier report, it didnot induce flowering under short-day conditions. (Received January 10, 1976; )     

Photoperiodic requirements for flowering of the long-day duckweed, Lemna gibba G3

The min-LD estimation by the log. flower number vs. the cultureperiod curve provides a unique method of judging whether a givenphotoperiodic schedule is a long day or not for Lemna gibbaG3. Duckweeds in M-sucrose medium are exposed to the test scheduleat 26°C on either the first or second day of a continuouslight culture. If the min-LD (48 hr for control cultures) isnot changed, the inserted schedule is considered to have functionedas a long day. If, however, the min-LD is extended by 24 hr,the inserted schedule is judged to have functioned as a shortday. Examinations using this method of orderly designed light-darkschedules disclosed two critical phases in the light requirement;the initial and terminal 1 hr portions (designated the L1- andL2-phases) of the subjective day. Thus, a given day became along or short day when both the L1- and L2-phases were illuminatedor when either or both of the two phases were darkened. Thecritical daylength (11.5 hr) was just long enough to cover boththe L1- and L2-phases and the inductive phase (L2-phase) waslocated at the end of the subjective day. (Received June 9, 1975; )     

Flowering of the long-day plant, Lemna gibba, under short-day schedules composed of red and far-red light

Flowering in Lemna gibba, a long-day duckweed, can be inducedunder a short-day condition when the photoperiodic regimes areR₇FR₃ (7 hr red followed by 3 hr far-red), R₅FR₅ and R₃FR₇.This indicates the necessity of a proper balance between redand far-red effects for flowering. The flowering induced bythese regimes is inhibited by a brief exposure to red givenat the start of darkness and this inhibition is reversed bysubsequent exposure to far-red. Thus, the red/far-red reversibleeffect is found only at the beginning of darkness for floweringof L. gibba. However, flowering of L. gibba is promoted by a red light breakgiven near the middle of a 14 hr dark period. The promotiveeffect is not reversed by subsequent exposure to far-red, i.e.,the effect of the red break converts from inhibition to promotionas when given later in the dark period, which suggests the involvementof a timing mechanism. (Received July 21, 1973; )     

Removal of the sugar inhibition of flowering in Lemna gibba G3 by catecholamines

DL-Epinephrine (10^–8–10^–7 M), DL-norepinephrine(10⁶ M) and DL-isoproterenol (10^–8–10^–6 M)alleviated floral inhibition due to 1% of sucrose, in Lemnagibba G3. The induction period extended by sucrose was curtailedby epinephrine, frond multiplication enhanced by the sugar beingleft unaltered. The pattern of action of catecholamines appearedto be very similar to that of cAMP. DL-Epinephrine, however,was ineffective in the presence of 10^–7 M DL-propranololwhich affected neither flower nor frond production by itself.Quabain and nicotinic acid also nullified the epinephrine actionon duckweed flowering. These and other relevant findings supportthe hypothesis that the cAMP-adenyl cyclase system participatesin the processes of flower induction in this long-day plant. (Received August 21, 1973; )     

Effect of light on the metabolism of thymidine in the long-day duckweed, Lemna gibba G3

Metabolism patterns of exogenous thymidine as disclosed by ECTEOLAcellulose column chromatography, were examined with a long-dayduckweed, Lemna gibba G3, under dark and light conditions. Whenthymidine-6-³H was applied, the pattern of thymidine metabolismin the light was not very different from that in the dark. However,when thymidine (methyl-³H) was used, incorporation of radioactivityinto two major ( and ß) and one minor components ofboth B and D fractions separated by column chromatography, wasstrikingly stimulated by the light, through photosynthetic activity.Component a of fraction B was tentatively concluded to be ß-ureidoisobutyricacid by paper chromatography. As the radioactivity from thymidine-6-³Hwas hardly recovered in the a component of fraction D, the partof the thymidine molecule incorporated into this component wasnot the pyrimidine ring, but a methyl residue. (Received March 29, 1973; )     

Replacement by ionophores of the photoperiodic light requirement in Lemna gibba G3

Light requirement during the phytochrome-mediated L1-phase ofLemna gibba G3 can be replaced by 10 µg/liter of valinomycinor gramicidin, supporting the idea (5) that a promoted K⁺-iontransport across membranes would be involved in the photoperiodiclight action on the L1-phase. (Received June 6, 1977; )     

Removal by chemicals of photoperiodic light requirements of Lemna gibba G3

Both the initial and the terminal 1 hr portions of the subjectiveday fraction, namely the L1- arid L2-phases, of a 24 hr daymust be illuminated in order for the day to be perceived asa long day in the min-LD determination by the long-day plant,Lemna gibba G3 (9). The light requirement of the L1-phase wassatisfied by a 10 min red light pulse given at the beginningof the phase. The red light effect was erased by a subsequent10 min far-red light, indicating phytochrome-mediated processesoccurring in the L1-phase. The light requirement of the L2-phasewas satisfied by blue or far-red light given during the terminal10 min period of the phase; there was no indication of phytochromeinvolvement. The light action on the L1-phase was replaced by10^–5 M of cyclic AMP or 10^–7 M of DL-isoproterenol.The isoproterenol action was antagonized by 10^–7 M ofDL-propranolol. Cyclic AMP (10^–5 M) combined with salicylicacid (10^–6 M), which can remove the light requirementof the L2-phase (10), rendered a completely dark day physiologicallyequivalent to a long day. Acetylcholine (10^–5 M) exertednyctomimetic action on the L1-phase of the second light day.The action of acetylcholine was antagonized by cyclic AMP (10^–5M). The L2-phase required no light in the presence of 10^–7M of DL-propranolol, and this propranolol action was not affectedby isoproterenol. These findings suggest changes in membranepermeability caused by the light given during the L1- and L2-phases. (Received July 7, 1976; )     

Comparison of the flowering behavior of the long-day plant Lemna gibba G3 from different laboratories

In an effort to determine the cause for the wide discrepanciesin the level of flowering response reported for the long-dayplant Lemna gibba L., strain G3, cultures of L. gibba G3 wereobtained from the laboratories of W. S. Hillman (G3-H), R. Kandeler(G3-K), Y. Oota (G3-O) and and A. Pieterse (G3-P) and comparedto the L. gibba G3 (G3-C) from this laboratory. Under continuouslight all cultures gave FL% values of 77 or above, and on a9L:15D short-day treatment, all cultures were completely vegetative.However, on daylengths of 10 to 12 hr, small but statisticallysignificant differences were obtained for the different cultures.The critical daylength curves for G3-G, which showed the shortestcritical daylength, and G3-K, which showed the longest criticaldaylength, differed by approximately one hour. Salicylic acidtreatment caused flower promotion in each culture, but statisticallysignificant differences were obtained between some of the culturesin their response to salicylic acid. It is concluded that the large discrepancies in the floweringresponses of L. gibba G3 that have been reported are due primarilyto differences in culturing methods and counting proceduresin the different laboratories. However, the results also indicatethat there may be distinct cultures of L. gibba G3 that exhibitsmall physiological and/or genetic differences that would makeprecise quantitative comparison between different laboratoriesvery difficult. (Received January 23, 1979; )     

Effects of some metabolic inhibitors on flowering of Lemna gibba G3, a long-day duckweed

Flowering of Lemna gibba G₃, a long-day duckweed, was inhibitedby adding CuSO₄, AgNO₃, HgCl₂, Na₂WO₄ or iodoacetamide to themedium at the concentrations inducing long-day flowering inLemna paucicostata 6746, a short-day duckweed. This suggeststhat these metabolic inhibitors affected the photoperiodic sensitivityrather than directly affecting flower initiation. Ferricyanidepromoted flowering in both of these short-day and long-day duckweeds. (Received July 7, 1977; )     

Maintenance of high Pfr-level in the dark period in relation to flowering in Lemna gibba G3

The relative Pfr-level in a long-day duckweed, Lemna gibba G3,was estimated by the null response method. The null % R value(% R in a R/FR-mixture that provides a null flowering response.This value was assumed to indicate the endogenous Pfr-levelof the duckweed.) remained high during the initial hours ofthe 15 hr nyctoperiod then decreased rapidly, if a 12 or 33hr photoperiod preceded the nyctoperiod. The null % R valuedropped immediately after the start of the 15 hr nyctoperiodsubsequent to a 1 or 24 hr photoperiod. Thus, the duration ofthe maintenance of a high Pfr-level changed rhythmically dependingon the length of the preceding photoperiod. Nyctoperiods ofup to 9 hr following a 12 hr photoperiod hardly affected flowering,but nyctoperiods given after a 24 hr photoperiod suppressedflowering in proportion to the length of the period. The Pfr-levelin the nyctoperiod, therefore, seems to be important for flowering,and phytochrome change, as a function of the length of photoperiod,may serve as a photoperiodic timer. Although floral responseto interruption with R or FR changed with the application period,the difference in response between R-treated and FR-treatedplants was relatively constant during a 15 hr nyctoperiod combinedwith a photoperiod of any length other than 1 hr. Apparently,the floral response to the R or FR pulse was regulated by ashift in the Pfr-level caused by the light pulse. (Received April 2, 1979; )     

Investigations on the effects of metal ions and chelating agents on growth and flowering of Lemna gibba G 3

The effect of different chelating agents on growth and floweringof Lemna gibba G 3 was studied in M and HUTNER'S media. Theincorporation of EDDHA and Fe-EDDHA in the media resulted inprofuse flowering and gibbous character of the fronds. However,EDTA was relatively less effective. It was demonstrated thatthe metal which influenced flowering in L. gibba G 3 was mostlikely copper. (Received August 19, 1970; )     

The occurrence of acetylcholine in Lemna gibba G3

The occurrence of acetylcholine in a long-day duckweed, Lemnagibba G3 has been demonstrated. After a preliminary purificationof the formic acid-acetone extract by Sephadex G-15 column chromatography,acetylcholine was identified by paper chromatography, pharmacologicalactivity on frog muscle, and sensitivity to acetylcholinesterase. Acetylcholine contents relative to that at the start of theexperimental culture were 0.99?0.06, 1.61?0.27, and 1.17?0.16after 2 cycles of the [9(15)], [16(8)], and [24(0)] schedules,respectively. (Received November 14, 1977; )     

Flowering Responses of the Long-day Plant Lemna gibba G3

Lemna gibba L., strain G3, exhibits a qualitative long-day flowering response with a critical daylength on a 24-hour cycle of about 10 hours. Evidence is presented that the onset of daughter frond formation in a given frond inhibits the activity of the flowering meristem. Consequently, flower induction can only occur in fronds smaller than about 0.05 to 0.07 mm long. Although a minimum of 1 long day seems to be sufficient to induce the formation of flower primordia, at least 6 long days are required to obtain mature flowers since long days are also required for the early stages of flower development. The critical night length on 24, 48 and 72-hour cycles is respectively 14, 16, and 18 to 22 hours. The close similarity between the critical night length for the different cycle lengths is explained in terms of an inhibitory effect of darkness both on flower initiation and flower development. A 10-hour dark period is more inhibitory to flowering on a 36-hour cycle than on 24, 48, 60 or 72-hour cycles. It is suggested that darkness inhibits flowering through the formation of a light-labile flower inhibitor which acts to inhibit the functioning of the flowering stimulus.