Long-day flowering of Lemna perpusilla 6746 in Mo-deficient medium

Lemna perpusilla 6746, a short-day duckweed, flowered undercontinuous illumination if some of the SH inhibitors, such ascyanide or tungstate were added to the M-sucrose medium. Theeffect of tungstate was not overcome by simultaneous applicationof molybdate, but deletion of the Mo from the medium was enoughto induce the long-day flowering. In vivo assay of nitrate reductaseactivity suggested that nitrate reduction was not inhibitedby tungstate, CuSO₄ or AgNO₃ which induced longday flowering.The possibility was suggested that suppression of some Mo-requiringprocess other than nitrate reduction brings about the long-dayflowering in this plant. (Received November 12, 1975; )     

Effect of Ferricyanide, Ferrocyanide and KCN on Growth and Flowering in the Short-Day Plant Lemna paucicostata 6746

Flowering in the short-day plant Lemna paucicostata 6746 canbe induced under continuous light by the addition of ferricyanie,ferrocyanide or KCN to M-sucrose medium. Each substance is nearly10 times more effective when the flasks are covered by glassbeakers than when cotton plugs are used. By contrast, when floweringis induced under continuous light by copper or by short-daytreatment, neither flowering nor growth are affected by whetherglass beakers or cotton plugs are used. Ferricyanide, ferrocyanideand KCN are also able to induce long-day flowering when theplants are grown on Msucrose medium in small beakers that areplaced in a covered storage dish that also contains a solutionof one of these compounds. Addition of a KOH trap to the storagedish completely blocks the flowering induced by these compounds.If [¹⁴C]ferrocyanide is added to the storage dish both the M-sucrosemedium and the plants contain significant amounts of radioactivity,the amount of radioactivity being proportional to the floweringresponse. These results indicate that ferricyanide, ferrocyanideand KCN break down to release HCN and that it is the HCN whichis responsible for inducing flowering in L. paucicostata 6746under continuous light. ¹Present address: Department of Biology, Osaka Kyoiku University,Ikeda, Osaka 563, Japan. ²Present address: Institute of Horticulture, The Volcani Center,P. O. B. 6, Bet-Dagan, Israel. (Received January 17, 1983; Accepted March 24, 1983)     

Effects of some SH-inhibitors and EDTA on flowering in Lemna perpusilla 6746

Lemna perpusilla 6746, a short-day duckweed, flowered undercontinuous illumination on M-sucrose medium containing CuSO₄,AgNO₃ and HgCl₂, which are SH-inhibitors. The optimum concentrationsof CuSO₄, AgNO₃ and HgCl₂ were 5, 1 and 20 µM, respectively.Other metal ions tested were ineffective, but at least two otherSHinhibitors, potassium ferricyanide and iodoacetamide, alsoinduced long-day flowering at the concentrations of 0.1-1 µM. Adding 50 µM EDTA to the medium prevented the effect ofcupric ion, but not that of other SH-inhibitors. EDTA at 200µM induced some long-day flowering when added to a mediumwith no SH-inhibitors. It also permitted some flowering whenadded together with cupric ion, and accelerated flowering inthe presence of the other SHinhibitors listed above. EDTA andSH-inhibitor effects appeared to be additive. (Received May 25, 1973; )     

The response of Lemna gibba G3 to a single long day in the presence of EDTA

Lemna gibba G3 in M-EDTA or M-EDTA-sucrose medium initiatesflowers in response to a single long day, but requires a minimumof nearly or exactly 2 long days in an M or M-sucrose mediumfor flower induction. (Received December 18, 1971; )     

Flower Induction by Suppression of Nitrate Assimilation in Lemna paucicostata 6746

In vitro activity of nitrate reductase was studied in Lemnapaucicostata 6746 grown on modified Hoagland medium supplementedwith 1% sucrose, containing various inhibitors. Copper, silver,tungstate or cyanide which induces daylength-independent flowering,inhibited the nitrate reductase activity, but azide which doesnot induce daylength-independent flowering did not. Molybdate-deficientmedium induced flowering, and inhibited nitrate reductase activity.Lowering of nitrate level of the medium also induced daylength-independentflowering. These results suggest that the suppression of nitrate assimilationcauses daylength independent flowering in Lemna paucicostata6746, and that one of the flower-inducing actions of the copper,silver, tungstate, cyanide or the deletion of molybdate is tosuppress the nitrate assimilation. (Received June 26, 1985; Accepted October 30, 1985)     

Flower-Inducing Activity of Lysine in Lemna paucicostata 6746

Flowering of Lemna paucicostata 6746, a typical short-day plant,was induced by culture for 96 or 120 h in nitrogen-free mediumunder continuous illumination. To examine the effects of lysine,we homogenized entire plants of L. paucicostata 151 in a solutionof lysine and the supernatant obtained after centrifugationof the homogenate was added to the medium to give various concentrationsof lysine in the medium. Flowering of strain 6746 in nitrogen-freeor nitrogen-deficient culture medium was effectively promotedby the addition of a lysine-containing supernatant to the medium.The suppressive effect of elastatinal, a protease inhibitor,on the induction of flowering was almost completely reversedby the simultaneous application of a lysine-containing supernatantto the medium. During nitrogen-free culture, the level of endogenousfree lysine, expressed on the basis of the amount of total freeamino acids, increased. Lysine-containing supernatants alsoinduced flowering of plants in nitrogen-rich medium under continuousillumination. These findings suggest that endogenous lysineis involved in the induction of flowering in L. paucicostata6746 on nitrogen-free or nitrogen-deficient medium, as it isin the induction of flowering in L. paucicostata 151 (Received July 29, 1996; Accepted November 18, 1996)     

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; )     

Flower-inducing Effect of Benzoic and Salicylic Acids in Various Strains of Lemna paucicostata and L. minor

The flower-inducing activities of benzoic and salicylic acidsadded to the medium differ with the species (Lemna paucicostataand L. minor), and even with the strains used. The type andpH of the medium used, full or 1/10 strength M medium at pH3.8, 4.4 or 5.1, or 1/2 or 1/20 strength NH₄⁺-free Hutner'smedium at pH 5.0, 6.0 or 7.0, also modify their activity. L.paucicostata, strain 151 is the most sensitive of the strainsused to both benzoic and salicylic acids followed by strain381. Such dramatic flowering responses were not obtained withthe other strains, but even strain 321, reportedly insensitiveto benzoic acid, could be induced to flower by adding benzoicacid to a modification of the medium. Benzoic acid is more effectivethan salicylic acid for all strains of L. paucicostata, butthe contrary is true for two L. minor strains tested. A higherpercentage of flowering is obtained in L. paucicostata in 1/2strength NH₄⁺-free Huter'sn medium than in M medium, exceptfor strain 151. When diluted, both media enhance flowering inall L. paucicostata strains. Generally, a lower concentrationof benzoic acid or salicylic acid is enough to induce floweringwhen the pH of the medium is lower. (Received March 30, 1981; Accepted May 16, 1981)     

Inhibition of Flowering in the Long-Day Plant Lemna gibba G3 by Hutner's Medium and its Reversal by Medium Modification

The long-day plant Lemna gibba G3 flowers normally in E medium(Hoagland-type medium plus 30 µM EDTA) but in 0.5 H mediumthere is no flowering. Ammonium is present in 0.5 H medium andis known to inhibit flowering in L. gibba G3, but even in NH₄⁺-free0.5 H medium there is virtually no flowering under continuouslight. Increasing the phosphate concentration of the NH₄⁺-free0.5 H medium from 1.15 ITIM to 12 or 16 mM results in substantialflowering. Decreasing the EDTA concentration from 850 µIMto 250 µM, or raising the nitrate concentration from 4mM to 12 mM, results in only a small increase in flowering.If the decrease in EDTA and increase in nitrate are combinedwith the increase in phosphate, however, the flowering responseis nearly as good as that obtained using E medium. Thus, withthese three changes the inhibitory effect of NH₄⁺free 0.5 Hmedium for flowering in L. gibba G3 is almost completely reversed In the above studies flowering was not limited by daylength.When plants were grown on E medium under an 11 hour daylengthwhere flowering is limited by daylength, decreasing the phosphateconcentration in the medium reduced flowering, but increasingthe phosphate concentration in the medium did not stimulateflowering. Thus, when flowering is limited by daylength, highphosphate will not cause flowering, but a certain level of phosphateappears to be necessary for the expression of photoinductionunder long days. (Received January 14, 1986; Accepted June 24, 1986)     

Growth and flowering of Lemna paucicostata I. General aspects and role of chelating agents in flowering

Lemna paucicostata HEGELM. is normally a short-day plant andflowers only in the presence of a chelating agent (EDTA or EDDHA)in the medium. The plant can be induced to flower even by asingle long night treatment; the flowering percentage, however,increases with further inductive cycles. The length of the criticaldark period depends upon the chelating agent employed in themedium. It is between 10 and 12 hr in the medium containingEDTA and about 8 hr in the EDDHA-supplemented medium. Red lightinterruption in the middle of the dark period—even fora minute—is inhibitory for flowering. Attempts to identify the metal ion(s) chelated reveal that thechelating agents affect flowering by facilitating iron uptake.This is also supported by the fact that the requirement of achelating agent for flowering can be overcome with an excessof iron in the medium. Interestingly, provision of EDDHA andexcess of ferric citrate, together, can bring about floweringeven under long days. ¹Originally HEGELMAIER (1) designated L. paucicostata as a separatespecies; however, THOMPSON (2) and DAUBS (3) have treated itsynonymous to L. perpusilla. More recently, based on physiologicaland chemotaxonomic studies, the distinctiveness of L. paucicostatafrom L. perpusilla has been favoured (4, 5). (Received September 8, 1969; )     

Flower Induction by Nitrogen Deficiency in Lemna paucicostata 6746

Lemna paucicostata 6746, a short-day plant, produced flowerbuds even under continuous light when cultured in nitrogen-deficientmodified Hoagland medium with 1% sucrose for 3 days or morefollowed by culture on nitrogen-rich medium (either nitrateor ammonium). Flowering was also induced by culture on mediumcontaining 20–100 µM nitrate as the sole nitrogensource for 10 days or more, but not on medium with a low ammoniumconcentration. However, if plants cultured on medium containing5–20 µM ammonium as the sole nitrogen source for10 days were grown in a nitrogen-rich medium for a further 4days, they produced flower buds. Thus, nitrogen deficiency caninduce day length-independent flowering in Lemna paucicoslata6746, but nitrogen is required for the manifestation of flowering. (Received January 31, 1986; Accepted April 24, 1986)     

Enhancement of long-day flowering by Mo-deficiency and application of some amino acids and asparagine in the short-day plant Lemna paucicostata 6746

The short-day plant Lemna paucicostata 6746 can be induced toflower on long days (continuous light) by the addition of copper,tungstate or ferricyanide to the medium, and in each case theeffect was greatly enhanced by deleting molybdate from the medium.Treatment with asparagine, aspartate, glutamate, -alanine, glycineorserine, all of which are known to increase the critical daylengthand almost nullify the light-break effect in L. paucicostata6746, enhanced the long-day flowering induced by copper, ferricyanideor Mo-deficiency. (Received October 13, 1978; )     

Flowering Induced by Nitrogen Deficiency in Lemna paucicostata 151

Lemna paucicostata 151 is a weakly responsive short-day plantwhen grown in non-aseptic ten-fold diluted E medium supplementedwith 1 µM 6-benzyladenine, but it flowered even underlong-day conditions (continuous light) when grown in this mediumfor more than 14 days. On the 14th day of culture, the levelof endogenous nitrogen in the plants decreased to about 60%of that in the plants inoculated at the start of the culture.The flowering obtained under long-day conditions was suppressedby raising the concentration of nitrogen in the medium, whileit was induced more rapidly by lowering the concentration ofnitrogen in the medium. Benzyladenine did not cause floweringby itself, but it was required for the flowering that was inducedby a reduction in the level of nitrogen in the medium. Thus,the flowering observed under long-day conditions is due to nitrogendeficiency in the plants. Two inhibitors of proteases, bestatin and elastatinal, clearlyinhibited the flowering induced by nitrogen deficiency. It appears,therefore, that the induction or activation of some protease(s)is involved in the flowering that is induced by nitrogen deficiency. (Received March 19, 1991; Accepted August 16, 1991)     

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     

Flowering responses of Lemna perpusilla and L. gibba in relation to nitrate concentration in the culture medium

Flowering responses of Lemna perpusilla strain 6746, a short-dayplant, and L. gibba strain G3, a long-day plant, to nitrateconcentration in Hoagland's type medium with or without EDTA,were compared. Maximum flowering of L. perpusilla under SD occurredat higher nitrate concentrations than did colony proliferation.Even under CL, L. perpusilla grown at sub-optimal nitrate concentrationsfor colony proliferation, flowered irrespective of the presenceof EDTA which reduces flowering. Unlike L. perpusilla, L. gibba failed to flower under SD atany nitrate concentration whether or not EDTA was added. UnderCL, however, L. gibba flowered at almost any nitrate concentrationwith or without EDTA. Double optima for nitrate concentrationwas exhibited in the presence of EDTA; optimal concentrationfor colony proliferation came between the two optima for flowering. We concluded that the nitrogen level of the medium is importantin regulating flowering of duckweeds, and that the effect ofEDTA, if any, may primarily be on colony proliferation and onlysecondarily or antagonistically on flowering. ¹ Present address: Institute for Agricultural Research, TohokuUniversity, Sendai 980, Japan. (Received September 25, 1971; )     

A Comparative Study on the Short-Day- and the Benzoic Acid-Induced Flowering in Lemna paucicostata

The addition of a high concentration of FeCl₃ to the medium(1/10 strength M medium) slightly inhibited the benzoic acid(BA)-induced flowering of Lemna paucicostata 151 in continuouslight, although it promoted the flowering induced by short-day(SD) conditions. SD treatment had no significant effect on BA-inducedflowering in the medium with a standard concentration of iron,in which this plant hardly responds to SD, but greatly promotedit in the medium containing iron at a high concentration, inwhich this plant clearly responds to SD. The effect of BA seemsto be independent of but additive to the photoperiodic stimulus. In photosensitive strains 6746 and 441, a low concentrationof BA slightly lengthened the critical photoperiod but had noflower-inducing effect under continuous light. Since an optimalconcentration of BA induced flowering even under continuouslight in these strains, this was considered to be due to photoperiod-independentpromotion of flowering rather than shortening of the time-measuringprocess in the photoperiodic reaction. (Received August 19, 1986; Accepted February 21, 1987)     

Absorption of Copper by Lemna as Influenced by Some Factors Which Nullify the Copper Effect on Flowering and Growth

The effect of copper on flowering and growth of Lemna paucicostata6746 and Lemna gibba G3 in a copper-containing medium is nullifiedby the addition of EDTA, ammonium ions or salicylic acid tothe medium or a decrease in its nitrate concentration. Thesefactors were examined for their effects on the absorption ofcopper by the plants. The addition of EDTA to the medium completelyinhibited the absorption of copper in both species, thus eliminatingthe copper effect. Ammonium ions also inhibited copper absorption,their effectiveness rising with their concentration. Loweringthe nitrate concentration in the medium nullified the coppereffect on flowering in L. paucicostata 6746, and the additionof salicylic acid to the medium also nullified the copper effectin L. gibba G3, both without affecting the absorption of copper. (Received June 7, 1982; Accepted August 27, 1982)     

Flower-Inducing Activity of Lysine in Lemna paucicostata 151 Cultured on Nitrogen-Deficient Medium

Lemna paucicostata 151, a weakly responsive short-day plant,flowers even under continuous illumination when cultured onnitrogen-free medium for more than 72 hours with subsequentculture on nitrogen-rich medium. During the nitrogen-free culture,the protease activity and protein content of the plant increasedand decreased, respectively. The plant contained a protein(s)that induced flowering of the plant when added to the medium.The level of this protein(s) also decreased during the nitrogen-freeculture. The total amount of free amino acids in plants culturedon nitrogen-free medium for 96 hours decreased to about 15%of that in plants at the start of nitrogen-free culture, butlevels of some amino acids increased. These amino acids wereexamined for their effects on flowering of plants cultured onnitrogen-deficient or nitrogen-free medium. Most of the aminoacids had no effect on flowering. However, profuse floweringwas induced when lysine was added to the medium. Lysine promotedthe flower-inductive process(es) rather than the developmentof flower buds. These results suggest that nitrogen deficiency-induced floweringof the plants is induced by lysine, which is generated froma specific protein(s) by proteolysis. (Received May 11, 1992; Accepted July 30, 1992)     

Measurement of Endogenous Phosphorus Levels in Relation Flowering in the Long-Day Plant Lemna gibba G3

When Lemna gibba G3 was grown on E medium, a decrease in thephosphate concentration caused a suppression of flowering andlead to a decrease in the phosphorus concentration in the plants.Addition of salicylic acid reversed this inhibition withoutcausing an increase in the phosphorus concentration, while additionof copper or ammonium to Hoagland-type medium inhibited flowering,also without affecting the phosphorus concentration. Plantsgrown under 8 h or 24 h daylengths exhibited a FL% of 0 or about50, respectively, but showed no difference in their phosphorusconcentrations. These results indicate that a high phosphorusconcentration is not always required for flowering in Lemnagibba G3. When plants were grown on a modified E medium in which 1/50strength Hutner’s medium micronutrients were substitutedfor the normal E medium micronutrients, the suppression of floweringcaused by reduced phosphate was completely eliminated. Addingeach micronutrient individually at the normal concentrationto the modified E medium demonstrated that manganese inhibitedflowering in modified E medium with a low phosphate concentration. (Received January 31, 1986; Accepted July 4, 1986)     

Flower-inducing Activity of Vitamin K in Lemna paucicostata

Vitamins K₁ K₃ and K₅ induced flowering in Lemna paucicostata151, a short-day plant, cultured in 1/10 strength M medium (1/10M medium) under continuous light, and their activity was greatlyintensified by simultaneous application of benzyladenine. Themost active of these was vitamin K₅ L. paucicostata 6746 ismore sensitive to vitamin K₅ than strain 151, but the effectof vitamin K₅ on strain 6746 was not intensified by benzyladenine.The flower-inducing activity of vitamin K₅ was intensified bythe addition of benzoic acid in both strains and by the additionof copper or ferricyanide in Strain 6746, when these chemicalswere added at such low concentrations that they would scarcelyinduce flowering. In strain 6746, vitamin K₅ added to 1/10 M had little effecton flowering under a subcritical photoperiod, while it clearlyinduced flowering under continuous light. In this strain, vitaminK₅ added to full strength M medium, in which this plant wasmore sensitive to short photoperiods than in 1/10 M medium,did not induce flowering even under continuous light, and wasrather inhibitory under short photoperiods. (Received August 14, 1984; Accepted October 16, 1984)