Studies of the Involvement of an Endogenous Rhythm in the Photoperiodic Response of Hyoscyamus niger

An attempt was made to determine the involvement of an endogenous circadian rhythm in the flowering response of the long-day plant Hyoscyamus niger L. grown in a modified White's medium. Both variable-cycle-length and light interruption experiments were employed in this attempt. In the variable-cycle experiments, plants were subjected to light periods of 6, 12, or 18 hours followed by varying lengths of darkness. The total lengths of the cycles varied from 12 to 72 hours. In experiments utilizing a 6-hour photoperiod, a high level of flowering occurred in cycle lengths of 12, 36, and 60 hours. Flowering was suppressed in the 24-, 48-, and 72-hour cycles. When a 12-hour photoperiod was used the flowering response was low between 24 and 36 hours and flowering did not indicate a rhythmic response. When an 18-hour photoperiod was used, the flowering response was suppressed in the 36- and 60-hour cycles.

Light-break experiments were conducted to study further the flowering response in Hyoscyamus. These experiments consisted of a 6-hour main photoperiod followed by varying lengths of darkness to make cycles of 24, 48, and 72 hours. At given intervals the dark period was interrupted by 2-hour light breaks. In a 24-hour cycle, flowering was promoted when a light break was given at either the twelfth or eighteenth hour of the cycle. In a 48-hour cycle, flowering was strongly promoted by light breaks given near the beginning or at the end of the dark period. In a 72-hour cycle, light breaks given at the eighteenth, forty-second, and sixty-sixth hour of the cycle stimulated flowering as compared with light breaks given at the thirtieth and fifty-fourth hour. These results are indicative of the involvement of an endogenous rhythm in the flowering response of Hyoscyamus niger.

     

Photoreactions Controlling Flowering of Chrysanthemum morifolium (Ramat. and Hemfl.) Illuminated with Fluorescent Lamps

Flowering of chrysanthemum plants under short photoperiods, as is well known, is prevented when the plants are illuminated near the middle of the long night. Such illumination inhibits flowering whether it is given continuously or intermittently, and whether it comes from incandescent or from fluorescent lamps. We discovered, however, that fluorescent light applied intermittently (cyclically) throughout the entire 16-hour long night was far less inhibitory than when applied during only part of this dark period. By contrast, incandescent filament illumination is strongly inhibitory under these conditions. The cycles of fluorescent light usually lasted 15 minutes, 1.5 minutes of light followed by 13.5 minutes of dark. When such cycles were applied for only 12 hours, leaving 4 hours of uninterrupted darkness in each long night, inhibition of flowering was complete again.     

Photoperiodic induction of flowering in green and photobleachedChenopodium rubrum L. ecotype 184 - a short- day plant

Chenopodium rubrum L. ecotype 184 is a qualitative short-day plant with critical length of the night of eight hours that must be exceeded in order to flower: Five days after sowing, the plants were exposed to a various number of inductive cycles (14/10 h of däy/night cycle) to test the optimal photoperiodic conditions for flowering. In our experimental conditions the plants flowered with high percentage after more than four received inductive cycles, but there was no flowering below that. The plants grown on the herbicide Norflurazon (photobleached plants) showed different photoperiodic characteristics. There was negligible flowering of photobleached plants in the same experimental conditions as for the green ones.     

Jasmonates Inhibit Flowering in Short-Day Plant Pharbitis nil

The role of jasmonates in the photoperiodic flower induction of short-day plant Pharbitis nil was investigated. The plants were grown in a special cycle: 72 h of darkness, 24 h of white light with lowered intensity, 24-h long inductive night, 14 days of continuous light. At 4 h of inductive night the cotyledons of non-induced plants contained about two times the amount of endogenous jasmonates (JA/JA-Me) compared to those induced. A 15-min long pulse of far red light (FR) applied at the end of a 24-h long white light phase inhibited flowering of P. nil. The concentration of jasmonates at 2 and 4 h of inductive night in the cotyledons of the plants treated with FR was similar. Red light (R) could reverse the effect of FR. R light applied after FR light decreased the content of jasmonates by about 50%. Methyl jasmonate (JA-Me) applied to cotyledons, shoot apices and cotyledon petioles of P. nil inhibited the formation of flower buds during the first half of a 24-h long inductive or 14-h long subinductive night. Application of JA-Me to the cotyledons was the most effective. None of the plants treated with JA-Me on the cotyledons in the middle of the inductive night formed terminal flower buds. The aspirin, ibuprofen and phenidone, jasmonates biosynthesis inhibitors partially reversed the effect of FR, stimulating the formation of axillary and terminal flower buds. Thus, the results obtained suggests that phytochrome system control both the photoperiodic flower induction and jasmonates metabolism. Jasmonates inhibit flowering in P. nil.     

Floral Inhibition of Biloxi Soybean During a 72-hour Cycle

The inhibitory effect of light interruptions given during the photophobe phases of a 72-hour cycle was studied with Biloxi soybean [Glycine max (L.) Merr.]. The basic 72-hour cycle consisted of 8 hours of light followed by 64 hours of darkness and was repeated 7 times. Supplementary white light treatments given at the twenty-fourth and/or forty-eighth hour of the cycle (photophil phases) promoted the flowering levels of the controls and kept light treatments given at the most inhibitory points from inhibiting flowering completely. Such supplementary light treatments did not affect the time of maximum sensitivity to light interruptions. When 30-minute light breaks were used, maximum inhibition occurred at the 16-, 43-, and 63-hour points. The duration of the light breaks affected the time of maximum inhibition when given during the second photophobe phase. The time of maximum inhibition occurred earlier with 4-hour light breaks than with either 3-minute or 2-hour light interruptions.

Three-minute red light interruptions produced essentially the same effect as 3-minute white light interruptions. Such treatments inhibited flowering completely in the first photophobe phase, inhibited flowering to only a small degree in the second photophobe phase, and inhibited flowering to an intermediate degree in the third photophobe phase. Far-red light interruptions strongly inhibited flowering in the first photophobe phase, especially when given early in the dark period. Three minutes of supplementary white light given at the twenty-fourth or forty-eighth hour of the cycle partially overcame the inhibitory effect of far-red light. Four hours of supplementary white light at these times completely overcame the far-red inhibition.

     

Cyclic Mononucleotide-Gibberellin Interactions in the Flowering and Proliferation of the Long-Day Plant Lemna gibba G3

3′,5′-cAMP stimulates flowering of Lemna gibba G3 under inductive long-day conditions and enhances flower onset. 3′,5′-cAMP has no influence on frond production. 2′,3′-cGMP increases markedly the proliferation of fronds and inhibits flowering. The effect of 2′,3′-cGMP on frond multiplication is photoperiodically independent; under short-day conditions 2′,3′-cGMP replaces in fact the requirement for inductive long-day conditions. 2′,3′-cGMP increases the total amount of DNA per frond. This accumulation of DNA precedes by 2–3 days the 2′,3′-cGMP related increase in frond formation. The results are discussed in the light of the hypothesis that the active cyclic mononucleotides exert their effects on multiplication and flowering at the level of DNA.     

Evidence that photoperiodic, dark time measurement in Pharbitis nil involves a circadian rather than a semidian rhythm

Experiments were carried out to determine whether a semidian (12 h) rhythm in flowering response operates in Pharbitis nil as the basis for photoperiodic time measurement. The effect of 5 min far-red light followed by 85 min dark (FRD) given 4, 8,14 and 22 h before the end of a 48 h photoperiod on night-break timing and critical night length was determined. When given 4 h before the end of a 48 h photoperiod, an interruption with FRD advanced the phase of the circadian rhythm in the night-break inhibition of flowering. In contrast, earlier interruptions of the photoperiod had no effect on the phase of the rhythm. The critical night length was modified by FRD given 4 h (shortened) or 8 h (lengthened) before the end of the photo-period; when given at other times FRD did not alter the critical night length. The results are discussed in relation to the basis for photoperiodic timekeeping, with particular reference to suggestions for the involvement of a semidian rhythm. A circadian model based on the concept of limit cycles is described.     

Inhibition of stem growth and flower formation in Pharbitis nil with N,N-dimethylaminosuccinamic acid (B 995)

Summary In the short-day plant Pharbitis nil, strain Violet, flower formation is inhibited by application of the growth retardant N,N-dimethylaminosuccinamic acid (B 995) via the roots for a period of 24 hours prior to one inductive long night. Terminal flower bud formation is suppressed by a B 995 concentration of 100 mg/l, but for complete suppression of all axillary flower buds 2000 mg/l is required. Inhibition of flower formation is also caused by B 995 application via plumules or cotyledons, even if made at the end of the inductive night. B 995 treatment always results in short, thick internodes and dark-green leaves.Transport of ¹⁴C-labeled B 995 from cotyledons to plumules and roots takes place during a 16-hour dark period. However, very little label moves from a treated to an untreated cotyledon. Application of B 995 to one of the two cotyledons results in flower inhibition, although the untreated cotyledon produces sufficient flower hormone to induce optimal flower formation. It is concluded therefore that in the short-day plant Pharbitis B 995 does not affect flower hormone production, but rather inhibits floral initiation by interfering with the action of the hormone in the shoot apex.Inhibition of flower formation by B 995 can be completely overcome by application of gibbrellin A₃ to the plumulus before the long nigh. A dose of 0.01 g GA₃/apex is sufficient to re-establish flowering, but much more GA₃ is required to restore internode length equal to that of the control. Indole-3-acetic acid and naphthalene acetic acid are totally inactive in overcoming B 995 inhibition of flower formation and growth.The growth rate of Pharbitis plants treated with B 995 and continuously grown in long-day conditions is initially low, but reaches the same level as in untreated plants approximately 25 days after treatment. ¹⁴C-labeled B 995 applied to cotyledons accumulates to a high degree in roots and in the basal part of the shoots. ¹⁴C-B 995 is metabolized very slowly and persists therefore in Pharbitis plants for prolonged periods of time.     

Effect of Low-intensity Red and Far-red Light and High-intensity White Light on the Flowering Response of the Long-day Plant Lemna gibba G3

The long-day plant Lemna gibba L., strain G3 exhibits a relatively low sensitivity to short, white-light interruptions given during the dark period of a short-day cycle. However, the plants are fairly sensitive to low-intensity red light treatments given during a 15-hour dark period on the third day of a 2LD-(9L:15D)-2LD-7SD schedule. Far-red light is almost as effective as red light, and attempts to reverse the red light response with subsequent far-red light treatments have not been successful. Blue light proved to be without effect. When plants were grown on a 48-hour cycle with 15 minutes of red light every 4 hours during the dark period, the critical daylength was reduced from about 32 hours to slightly less than 12 hours.

Continuous red light induced a fairly good flowering response. However, as little as 1 hour of white light each day gave a significant improvement in the flowering response over that of the continuous red light control. White light of 600 to 700 ft-c was more effective than white light of 60 to 70 ft-c. The white light was much more effective when divided into 2 equal exposures given 8 to 12 hours apart. These results suggest an increase in light sensitivity with regard to flower induction about 8 to 10 hours after the start of the light period.

     

Floral initiation in strawberry: Spectral evidence for the regulation of flowering by long-day inhibition

Summary Floral initiation in strawberry cv. Cambridge Favourite, a facultative short-day plant, was inhibited by a daylength extension with red light (R) during the second half of a 16-hour night but not during the first half, and by far-red light (FR) in the first half but not during the second. Mixed R plus FR light was inhibitory to flowering at both times. This change in sensitivity to R and FR light in the evening and morning resembles the pattern for flower induction in long-day plants but differs from the pattern for flower inhibition in several other short-day plants, examples of which are given. These experiments afford further support for the hypothesis that the control of flower initiation in strawberry depends on the production of a flower inhibitor by leaves exposed to long photoperiods.Abbreviations R red - FR far-red - SD short day - LD long day - SDP short-day plant - LDP long-day plant     

Inhibitory Effect of Methyl Jasmonate on Flowering and Elongation Growth in Pharbitis nil

The effect of methyl jasmonate (JA-Me) on the floral bud formation and elongation growth in the short-day plant Pharbitis nil was investigated. The placing of 4-day-old seedlings of P. nil in a solution of JA-Me for a period of 24 h before an inductive (16 h or 14 h of darkness) night led to a dramatic reduction in the number of flower buds formed by the plant. Plants treated with JA-Me also totally lost their capacity to form a generative terminal bud. JA-Me applied after photoinduction does not inhibit flowering. Gibberellic acid (GA3) partly reverses the inhibitory effect of JA-Me. Plants treated simultaneously with JA-Me and GA3 formed about 3 flower buds more than plants treated with JA-Me only. JA-Me at a concentration of 10-7 M stimulates slightly, but at higher concentrations it inhibits root growth and shoot growth. A distinct lack of correlation between the effect of JA-Me on inhibition of flowering and shoot and root growth was noted. This indicates the independent action of JA-Me in controlling both processes.     

EXPERIMENTS ON THE PHOTOPERIODIC RESPONSE IN PINEAPPLE

Gowing , Donald P. (Pineapple Research Institute of Hawaii, Honolulu.) Experiments on the photoperiodic response in pineapple. Amer. Jour. Bot. 48(1): 16–21. 1961.—The initiation of flowering of ‘Smooth Cayenne’ pineapple plants is neither strictly a response to photoperiod (day lengths of 10 hr. 51 min.–13 hr. 24 min.) nor to a minimum temperature (minima from 50° to 72°F. in different areas) under natural Hawaiian conditions. Depending on the kind of planting material used and the time of planting, natural initiation of flowering may take place any month of the year. Slips planted in the fall generally initiate flowering in December of the following year. However, exposure of an 8-mo.-old slip-planting to a day length of 8 hours for 40 days starting Sept. 8 induced flowering irrespective of night temperatures from about 60 to 80°F. Interruption of the dark period by illumination at 30 ft.-c. from midnight to 1 a.m. suppressed the inductive effect. Lowering the night temperature to 60°F. was, of itself, non-inductive. Field-grown, 11-mo.-old plants treated in place responded similarly, in that 25 periods of 8-hr. day length starting Sept. 5 induced 60% of the plants to flower, and the night illumination suppressed the inductive effect as before. Daily application of 0.12 mg. of the major native pineapple auxin (indole-3-acetic acid) at the beginning of the dark period had no detectable effect on the short-day treatment, and similar application of an antiauxin (4-chlorophenoxyisobutyric acid) did not affect the suppression of flowering by the light-break. Supplemental illumination of field-grown 12-mo. plants to provide a photoperiod of more than 15 hr. daily from Nov. 4 to Jan. 30 did not suppress the natural initiation of flowering which occurred in early December (day length about 10 hr. 50 min.). ‘Smooth Cayenne’ pineapple is therefore a quantitative, but not an obligate, short-day plant.     

Ethylene and ABA interactions in the regulation of flower induction in Pharbitis nil

Hormones are included in the essential elements that control the induction of flowering. Ethylene is thought to be a strong inhibitor of flowering in short day plants (SDPs), whereas the involvement of abscisic acid (ABA) in the regulation of flowering of plants is not well understood. The dual role of ABA in the photoperiodic flower induction of the SDP Pharbitis nil and the interaction between ABA and ethylene were examined in the present experiments. Application of ABA on the cotyledons during the inductive 16-h-long night inhibited flowering. However, ABA application on the cotyledons or the shoot apices during the subinductive 12-h-long night resulted in slight stimulation of flowering. Application of ABA also resulted in enhanced ethylene production. Whereas nordihydroguaiaretic acid (NDGA) - an ABA biosynthesis inhibitor - applied on the cotyledons of 5-d-old seedlings during the inductive night inhibited both the formation of axillary and of terminal flower buds, application of 2-aminoethoxyvinylglycine (AVG) and 2,5-norbornadiene (NBD) - inhibitors of ethylene action - reversed the inhibitory effect of ABA on flowering. ABA levels in the cotyledons of seedlings exposed to a 16-h-long inductive night markedly increased. Such an effect was not observed when the inductive night was interrupted with a 15-min-long red light pulse or when seedlings were treated at the same time with gaseous ethylene during the dark period. Lower levels of ABA were observed in seedlings treated with NDGA during the inductive night. These results may suggest that ABA plays an important role in the photoperiodic induction of flowering in P. nil seedlings, and that the inhibitory effect of ethylene on P. nil flowering inhibition may depend on its influence on the ABA level. A reversal of the inhibitory effect of ethylene on flower induction through a simultaneous treatment of induced seedlings with both ethylene and ABA strongly supports this hypothesis.     

Evidence for a Flowering Inhibitor Produced in Long Days in Kalanchoe blossfeldiana

Experiments with Kalanchoe blossfeldiana are described in whichperiods of short-day treatment were interrupted by intercalatedlong days or light breaks during long dark periods. The effectsof 24-hour dark periods preceding and following such intercalatedlong days were also investigated. The results of these experiments have shown that: Single longdays intercalated between numbers of short days have a positiveinhibitory effect on flower initiation and are not merely ineffective.The inhibitory effect expressed as the number of inductive cyclesannulled is approximately additive, provided the long days areinterspersed with short days, but not if several long days aregiven consecutively. On the average 1 long day is capable ofannulling the flower-promoting effect of about 1 short days.To a first approximation flower numbers in Kalanchoe increaseexponentially with the number of inductive cycles given—upto at least 12 short days; the inhibitory effect of long daysinterspersed with short days also fits an exponential curve;i.e. the inhibition is roughly proportional to the amount ofprevious photo-periodic induction. A light break of as littleas 30 seconds' duration given in the middle of a long dark periodis as inhibitory as a long day. If followed by a long dark periodthe inhibition of an intercalated long day is almost completelyneutralized; a long dark period preceding it has no such effect. These results have been interpreted as due to the interactionof a flowering inhibitor with a reaction leading to flowering.A mechanism involving competitive inhibition of an adaptivelyformed enzyme has been described as a possible example of thekind of reaction which could account for the results presented.     

The response of growth and circadian rhythm to cycle length and photoperiod

Summary The growth of bean plants (Phaseolus vulgaris L., cv Red Kidney) is inhibited by cycle lengths of 36 and 48 hours. Maximal inhibition occurs when the length of the light period is equal to or shorter than 3/6 of the cycle length. The inhibition does not occur when the photofraction is 5/6 or longer. The rhythmic leaf movement in beans can be entrained to a 30-hour cycle with a photofraction of 3/6 or less. No entrainment occurs to 36-or 48-hour cycles, but such cycles with photofraction of 3/6 or less cause an irregular course of the rhythm. When the photofraction is 5/6 or greater, the leaf movement proceeds as in continuous light, independent of cycle length. In continuous light the rhythm persists for at least 12 days. The parallel response of growth and circadian rhythm to cycle length and photoperiod suggests that a circadian oscillation is involved in the growth process. It further indicates that the response of these phenomena to cycle length and photoperiod involves the same basic timing mechanism.With 4 Figures in the Text     

Photoperiodic counter of diapause induction in Pseudopidorus fasciata (Lepidoptera: Zygaenidae)

Induction of larval diapause is a photoperiodically controlled event in the life history of the moth Pseudopidorus fasciata. In the present study, the photoperiodic counter of diapause induction has been systematically investigated. The required day number (RDN) for a 50% response was determined by transferring from a short night (LD 16:8) to a long night (LD 12:12) or vice versa at different times after hatching, The RND differed significantly between short- and long-night cycles at different temperatures. The RDN for long-night cycles at 20, 22, 25 and 28 degrees C was 11.5, 9.5, 7.5 and 8.5 days, respectively. The RDN for short-night cycles was 3 days at 22 degrees C and 5 days at 20 degrees C indicating that the effect of one short night was equivalent to the effect of 2-3 long nights effect. Night-interruption experiments of 24h photoperiods by a 1 h light pulse showed that the most crucial event for the photoperiodic time measurement in this moth was whether the length of pre-interruption (D(1)) or the post-interruption (D(2)) scotophases exceeded the critical night length (10.5 h). If D(1) or D(2) exceeded 10.5 h diapause was induced. The diapause-averting effect of a single short-night cycle (LD 16:8) against a background of long nights (LD 12:12) showed that the photoperiodic sensitivity was greatest during the first 7 days of the larval period and the highest sensitivity was on the fourth day. Both non-24 and 24 h light-dark cycle experiments revealed that the photoperiodic counter in P. fasciata is able to accumulate both long and short nights during the photosensitive period, but in different ways. The information from short-night cycles seems to be accumulated one by one in contrast to long-night cycles where six successive cycles were necessary for about 50% diapause induction and eight cycles for about 90% diapause. These results suggest the accumulation of long-night and short-night cycles may be based on different mechanisms.     

Circadian Rhythmicity in Photoperiodic Regulation of Regeneration in Begonia Leaves

The mechanism of photoperiodic regulation of regeneration in Begonia leaves has been studied by the night interruption technique in 24, 48, and 72-h cycles. The response to 30 min red light interruptions in 48 and 72-h cycles indicated a circadian rhythm in red light sensitivity with typical photophile and scotophile phases. In 24-h cycles two types of response patterns were observed. With a main photoperiod of 3 h the usual response pattern with only one light-sensitive phase near the middle of the dark period was found, whereas with 8-h photoperiods two light-sensitive phases were observed as previously reported by Zimmer in Begonia flowering studies (Gartenbauwissenschaft 38: 57, 1973). Reversion studies with FR indicate that the reactions are mediated by phytochrome. The results are discussed in relation to alternative hypotheses for photoperiodic timing.     

Independent effects of jasmonates and ethylene on inhibition of <Emphasis Type="Italic">Pharbitis nil</Emphasis> flowering

Interactions between methyl jasmonate (JA-Me) and ethylene in the photoperiodic flower induction of short-day plant Pharbitis nil were investigated. Both JA-Me and gaseous ethylene applied during the inductive long night caused a decrease in the number of flower buds generated by P. nil. Application of ethylene did not affected niether the level of endogenous jasmonates in the cotyledons during the 16 h long inductive night, nor the inhibitory effect of JA-Me on the flowering of P. nil accompanied by variations in ethylene production. The application of acetylsalicylic acid (aspirin)—a jasmonate biosynthesis inhibitor—slightly stimulated flowering. Our results have shown that the mechanisms of P. nil flower inhibition by jasmonates and ethylene are independent.     

Inhibitory Effect of Methyl Jasmonate on Flowering and Elongation Growth in Pharbitis nil

The effect of methyl jasmonate (JA-Me) on the floral bud formation and elongation growth in the short-day plant Pharbitis nil was investigated. The placing of 4-day-old seedlings of P. nil in a solution of JA-Me for a period of 24 h before an inductive (16 h or 14 h of darkness) night led to a dramatic reduction in the number of flower buds formed by the plant. Plants treated with JA-Me also totally lost their capacity to form a generative terminal bud. JA-Me applied after photoinduction does not inhibit flowering. Gibberellic acid (GA3) partly reverses the inhibitory effect of JA-Me. Plants treated simultaneously with JA-Me and GA3 formed about 3 flower buds more than plants treated with JA-Me only. JA-Me at a concentration of 10-7 M stimulates slightly, but at higher concentrations it inhibits root growth and shoot growth. A distinct lack of correlation between the effect of JA-Me on inhibition of flowering and shoot and root growth was noted. This indicates the independent action of JA-Me in controlling both processes.