5-Fluorodeoxyuridine inhibition of photoperiodically induced flowering inChenopodium rubrum L.

Flowering in the short day plantChenopodium rubrum was inhibited by 5-fluoro-deoxyuridine (FDU) at a concentration of 4×10^?6 M and higher when applied during photoperiodic induction or immediately afterwards. This inhibition is always accompanied by a general reduction of growth (e.g. a decrease in the first leaf length). The mitotic activity within the shoot apex is completely blocked by FDU application during the photoperiodic treatment. The floral induction (evocationsensu Evans) was not cancelled in this situation as was revealed when reversing the FDU effect by thymidine application. One day after the end of the photoperiodic treatment (the plants were transferred to continuous light again) the FDU inhibition becomes irreparable. The results indicate that DNA synthesis and hence the mitotic activity are not obligatory prerequisites for photoperiodic floral induction inChenopodium. Low concentrations of FDU may promote flowering under suboptimal floral induction.     

The incorporation of uridine-3H into the shoot apices of photoperiodically induced and non-induced plants ofChenopodium rubrum L.

The influence of photoperiodic induction on the incorporation of uridine-³H into the shoot apices ofChenopodium rubrum was studied using the technique of autoradiography. No increase in uridine incorporation was detected either during induction lasting three days or immediately after its termination. Pyroninophylia likewise did not rise. However, changes in uridine incorporation related to morphogenetic activity during leaf formation and later during differentiation of inflorescences were well marked. The distribution of label in the nucleus immediately after three inductive cycles shows the ratio of extranucleolar to nucleolar incorporation to be higher in non-induced control plants than in induced ones. Data from literature pointing to an activation of RNA synthesis during transition to flowering are discussed and compared with other systems where ontogenetic changes are accompanied by marked changes in RNA synthesis. It is assumed that the activation of RNA synthesis after induction is connected mainly with the activation of growth. However, inChenopodium rubrum photoperiodic induction proceeds together with limited growth and without activation of RNA synthesis.     

Changes in organ growth ofChenopodium rubrum due to suboptimal and multiple photoperiodic cycles with and without flowering effect

The growth changes of cotyledons, leaves, hypocotyls and roots due to photoperiodic induction in short day plantChenopodium rubrum were investigated in relation to flowering. Six-day old plants were induced by photoperiods with a different number of dark hours. We found that the degree of inhibition which occurred during induction in the growth of leaves, cotyledons and roots similarly as the stimulation of hypocotyl is proportional to the length of dark period. The photoperiods with 12, 16 and 20 dark hours bring about marked inhibition of growth and at the same time induce flowering in terminal and axillary meristems. The inhibitory effect of critical period for flowering,i.e. 8 dark hours, is not apparent in all criteria used and even the flower differentiation is retarded. The photoperiods of 4 and 6 dark hours did not affect growth and were ineffective in inducing flowering even if their number has been increased. The experiments with inductive photoperiod interrupted by light break have clearly shown that growth pattern characteristic for induced plants can be evoked in purely vegetative ones. Such statement did not exclude the possible importance of growth inhibition as a modifying factor of flower differentiation. We demonstrated that the early events of flower bud differentiation are accompanied by stimulation of leaf growth. The evaluation of growth and development of axillary buds at different nodes of insertion enabled us to quantify the photoperiodic effect and to detect the effects due to differences in dark period length not exceeding 2 hours.     

Inhibition of flowering ofChenopodium rubrum L. after the action of actinomycin D

Floral differentiation ofChenopodium rubrum is more AD-sensitive than growth of the vegetative organs. With a suitable combination of the manner of application and the concentration of AD used, selective inhibition of flowering can be attained without any effect on growth. The inhibition of flowering was greatest if AD acted during the first two days of photoperiodic induction. With later application its effect on flowering was weaker. RNA synthesized in the first days of photoperiodic induction to a considerable extent ensured its further course.     

Localization of starch in shoot apices of vegetative and photoperiodically induced plants ofChenopodium rubrutn

Starch was determined by means of IKI reaction in shoot apices ofChenopodium rubrum plants induced to flowering by two short days and in non-induced plants. Small starch grains were already observed in the meristematic cells at an age of four days after sowing. Larger grains were found in the subapical region of the apex. Heterogeneity increases during further growth of the plants in induced, as well as in non-induced vegetative plants. Starch disappears from the cells potentially giving rise to axillary buds, while the number and size of starch grains increase in cells from which leaf primordia will be formed. This metabolic specifity of leaf and bud primordia is preserved during morphological differentiation and applies to vegetative, as well as to prefloral apices of photoperiodically induced plants. The amount of starch in the different regions of the apex is linked rather with organogenesis than with the quantitative growth in the apex.     

Effect of different sugars on flowering ofChenopodium rubrum L. in dependence on the conditions of germination and initial growth

Flowering ofChenopodium rubrum seedlings fed different sugars at a concentration of 0.6 and 0.4 M, reap, during a single inductive cycle was stimulated or inhibited in dependence on the conditions of germination and initial growth. Plants allowed to germinate at alternating temperatures of 28 °C and 5 °C showed a slower initial growth and their development was stimulated by some sugars as compared to controls induced in the absence of sugars. Plants germinated at alternating temperatures of 32 °C and 5 °C exhibited a rapid initial growth and flowering was inhibited after induction in the presence of sugars. On the other hand, development proceeded more rapidly in control plants induced in the absence of sugars after germination at the higher temperature than after germination at the lower one. The differences between the two variants quoted above could be observed also after induction by two 16 h dark cycles. Glucose and sucrose were most effective in stimulating flowering under appropriate conditions of germination. Fructose was less effective and the action of maltose was very weak. Xylose, ribose and galactose were innocuous, while arabinose, glucoso-6-phosphate and mannitol were toxic to the plants. The sugars inhibited root growth in all cases and led to an increase in starch accumulation in the underground and overground plant organs. At a concentration of 0.6 M they mostly inhibited the length of the cotyledons and, especially, of the first leaf; at a concentration of 0.4 M growth of the overground organs was stimulated. The results are discussed with respect to the possible ohanges in photoperiodic sensitivity brought about by the rate of initial growth.     

Promotive effect of abscisic acid in flowering ofChenopodium rubrum as the result of decreasing apical dominance

Abscisic acid (ABA) was applied in a concentration of 1. 10^?3 M and 1. 10^?4 M to the quantitative SD plantChenopodium rubrum under various light regimes. ABA did not influence flowering in plants under continuous illumination, enhanced flowering in plants subjected to long days and inhibited it in plants induced by short days. It was concluded that ABA can not substitute for inductive treatment but its action may be additive to initial stages of reproductive morphogenesis (enhanced growth rate and branching of the apical meristem) as evoked by long days.     

Floral transition as a sequence of growth changes in different components of the shoot apical meristem ofChenopodium rubrum

The changes in cell division rate were studied in different components of the shoot apex ofChenopodium rubrum during short-day photoperiodic induction and after the inductive treatments. Induced and vegetative apices were compared. Accumulation of metaphases by colchicine treatment was used to compare the mean cell cycle duration in different components of the apex. A direct method of evaluating the increase in cell number obtained by anticlinal or periclinal divisions was applied if the corresponding components of induced and non-induced apices had to be compared. The short-day treatment prolonged the cell cycle more in the peripheral zone than in the central zone and still more in the leaf primordia. The importance of changing growth relations for floral transition was shown particularly if the induced plants were compared with the vegetative control with interrupted dark periods. Induced plants transferred to continuous light showed further changes in the rates of cell division. The cell cycle was shortened more in the central zone than in the peripheral zone,i.e. there was a further shift in growth relations within the apical dome. The cell cycle in the leaf and bud primordia was also shortened if compared with the vegetative control, the acceleration being stronger in the bud primordia. There was a subsequent retardation in cell division in the leaf primordia formed during and after the inductive treatment if the plants were fully induced. An inhibition of the oldest bud primordia was observed in fully induced apices, as well.     

Autoradiography of tritiated uridine in shoot apices ofChenopodium rubrum

The incorporation of uridine-5-³H into shoot apices ofChenopodium rubrum, plants was studied using autoradiography. The evaluation of the rate of incorporation into the nucleolus and the extranucleolar part of the nucleus as a function of the total radioactivity in the apex yields quantitative data on the distribution of labeling in these parts of the nucleus. Incubation of intact germinating plants in uridine-³H makes it possible to carry out chase experiments. Curves of uridine incorporation into the nucleolus and the extranucleolar part of the nucleus were obtained which demonstrated a non-linear course of incorporation. When incubating with uridine from 30 to 120 min the nucleolar/extranucleolar ratio of labelling was found to increase from 2 to 3. In chase experiments this ratio changed within three days from 3 to 1. Interpretation of these results in view of the function of RNA localized in different parts of the nucleus is discussed.     

The inhibition and stimulation of DNA synthesis in shoot apices ofChenopodium rubrum L. during photoperiodic induction of flowering

Three short-day inductive cycles bring about inhibition followed by transitional enhancement of growth, not only in roots and leaves but also in different zones of shoot apical meristem, as shown by measurement of DNA synthesis using³H-thymidine autoradiography. The first inductive cycle resulted in marked inhibition of the cells of the central zone (CZ), rib meristem (RM), and peripheral zone (PZ). Subsequent enhancement of DNA synthesis occurs in RM during the second inductive cycle, but in CZ only in the third cycle. The growth activation in PZ is counteracted by decrease in apical dominance which results in further inhibition of leaf primordia and increases in bud primordia. In plants induced only by one cycle, which later reverse the vegetative pattern of growth and differentiation, increased DNA synthesis in RM and CZ was not observed. The significance of inhibitory and stimulatory processes in particular zones of the shoot apex is discussed considering flower morphogenesis.     

Time-dependent reversal of the inhibitory effect of 5-fluorode-oxyuridine on flowering ofChenopodium rubrum L. by thymidine

Flower initiation induced by three inductive photoperiods inChenopodium rubrum L. was fully inhibited by treating the shoot apex with a 5 μl drop of 1×10^?5 m 5-fluorodeoxyuridine (FDU). This inhibition may be reversed by thymidine applied simultaneously with or after FDU treatment at any time during photoperiodic induction. One day after the end of induction the inhibition caused by FDU is irreparable even by increasing thymidine concentrations. It is concluded that photoperiodic floral induction may take place inChenopodium even if DNA synthesis is suppressed.     

Thyroxine can mimic photoperiodically induced gonadal growth in Japanese quail

Summary Pharmacological doses of thyroxine are able to mimic the effects of long photoperiods in Japanese quail. In birds maintained on short daylengths thrice-weekly injections of 100 g thyroxine cause full testicular maturation at rates not greatly different from those seen if quail are exposed to long days. Thyroxine stimulates increases in the secretion of FSH and LH, in pituitary prolactin content and in the hypothalamic content of Gn-RH. The effects are dose-dependent. If female quail kept on short daylengths are given thyroxine their ovaries develop and they lay eggs. In castrated male quail on short days thyroxine causes a ten-fold increase in circulating LH within a week. Thyroxine injections are also capable of maintaining quail in a photorefractory state even when they are transferred to short daylengths. The results suggest that thyroxine mimics long days by acting high in the photoneuroendocrine system and does not simply act to facilitate hormone secretion per se. This is in line with growing evidence in mammals and birds that parts of the photoperiodic machinery are sensitive to thyroid hormones.Abbreviations Gn-RH gonadotropin releasing hormone - FSH follicle stimulating hormone - LH luteinizing hormone - T ₄ thyroxine     

Growth correlations and rna synthesis in different parts of the shoot apical meristem ofChenopodium rubrunt L. induced to flowering

Uridine-³H incorporation and RNA concentration were investigated in different parts of the shoot apical meristem ofChenopodium rubrum using autoradiography and cytophotometry. A single inductive cycle was sufficient to bring about postinductive first events in the shoot apex but not for complete flower differentiation. The initial activation of RNA synthesis manifested itself in all zones of the apex. The first increase was more conspicuous in the peripheral than in the central zone. The indications of the first events in the apices after a single inductive cycle disappear prior to morphological reversal to the vegetative state. Induction by three short days led to rapid flower differentiation. The increase in RNA synthesis and concentration was most conspicuous in the central zone in this case. The ratio of RNA synthesis and content between bud and leaf primordia (B/L) also change in relation to photoperiodic induction. In vegetative plants the B/L ratio was low while after induction it increased. The shifts in activity of RNA synthesis observed in the shoot apical meristem are related to the changes in growth activity of the different parts of the apex. The growth ratios in the apices bear the character of growth correlations. The change in the growth correlations following photoperiodic induction together with the total activation of RNA synthesis are considered to represent one of the first events of the transition to the reproductive state.     

Rhythmic changes in ribonuclease activity in relation to nucleic acid synthesis in cotyledons ofChenopodium rubrum L. and to floral induction of the plants

Ribonuclease (RNAse) activity was investigated in cotyledons ofChenopodium rubrum plants subjected to various conditions of illumination (photoperiodic induction, continuous light, induction cancelled by interrupting the dark period by a light-break). At the end of the dark period of the single inductive cycles RNAse activity of induced plants was inferior to that of plants grown in continuous light. At the end of the first two cycles the activity was lowest after the interruption of the dark period by light. The investigation of the enzyme in 6h intervals showed rhythmic changes in activity to occur in induced plants. Enzyme activity followed a pattern opposed to this of nucleic acid (NA) synthesis in the cotyledons. In plants from continuous light the enzyme activity did not show any rhythm and in plants having obtained a light-break during the inductive period the rhythm was less distinct than in the induced ones. The period length of the endogenous rhythm of NA synthesis in the cotyledons is about half as long as this of flowering and the peaks of flowering coincide with the throughs of NA synthesis.     

Changes in the level of endogenous cytokinins in apical buds ofChenopodium rubrum L.

CCC (2-chloroethyl)trimethylammonium chloride applied to plants ofChenopodium rubrum during floral induction led to an increase in the level of endogenous cytokinins in the apical buds. Application of gibberellic acid or indole-3-acetic acid at concentrations reversing the effect of CCC reduced the level of cytokinins. After simultaneous treatment with both CCC and one of the growth substances this reduction was less pronounced. From the comparison bf the present results, as well as of those published in previous papers it follows that in apical buds ofChenopodium rubrum there exists a mutual interaction between gibberellins and cytokinins. Under certain conditions both these groups of hormones may substitute for each other in flowering. IAA seems to affect flowering by regulating the level of both gibberellins and cytokinins.     

Induction of flowering by 5-azacytidine in some plant species: relationship between the stability of photoperiodically induced flowering and flower-inducing effect of DNA demethylation

The flower-inducing effect of 5-azacytidine, a DNA demethylating reagent, was examined in several plant species with a stable or unstable photoperiodically induced flowering state under non-inductive photoperiodic conditions. The long day plant Silene armeria , whose flowering state is stable and the short day plant Pharbitis nil , whose flowering state is unstable were induced to flower by 5-azacytidine under a non-inductive condition. Thus, the replacement of photoinduction by 5-azacytidine treatment is not specific to Perilla frutescens . On the other hand, 5-azacytidine did not induce flowering in Xanthium strumarium whose flowering state is stable and Lemna paucicostata whose flowering state is unstable. Thus, epigenetics caused by DNA demethylation may be involved in the regulation of photoperiodic flowering irrespective of the stability of the photoperiodically induced flowering state.     

Endogenous abscisic acid in relation to photoperiodically induced bud dormancy

Summary The abscisic acid contents of birch, maple and sycamore plants growing under long and short photoperiods were measured by gas-liquid chromatography. No increase was observed in the abscisic acid content of extracts when plants were transferred to dormancy-inducing conditions.A preliminary account of this work was presented at the XI International Botanical Congress, 1969.     

Fluctuations of Uridine Incorporation in the Shoot Apex ofChenopodium rubrum L. during Photoperiodic Induction

Uridine incorporation into the shoot apex of the short-day plantChenopodium rubrum was investigated during a 16 h period of darkness and the following transfer to light. Uridine incorporation during this single inductive cycle was compared to incorporation under non-inductive conditions of continuous light. After transfer of the plants from light to darkness RNA synthesis was reduced to about half after the first two hours. This occurred not only when the plants were precultivated in continuous light but also after an interruption of the dark period by light for 31/2 h. The low level of uridine incorporation was maintained for the whole duration of the dark period. Incorporation regained its initial level after exposure of the plants to light irrespective of the duration of the preceding dark period. After this immediate rise of uridine incorporation in plants transferred from darkness to light a slight temporary decrease was observed in light. In darkness the decrease of incorporation into the nucleoli was still more marked than the reduction of overall incorporation. After the termination of the dark period incorporation into the nucleolus rose slowly and extranucleolar incorporation was relatively enhanced during the first 10 h of light in induced plants. The fluctuations of RNA synthesis observed in the shoot apex during photoperiodic treatment may be regarded as a necessary condition for the transition from the vegetative to the reproductive state.     

Development of the shoot apex ofChenopodium rubrum L. after photoperiodic induction in the cotyledon stage

Development of the shoot apex up to floral differentiation was investigated in the short-day plantChenopodium rubrum. The changes occurring in the apex from energence until full opening of the cotyledons (Figs 1–4), development during photoperiodic induction (Figs. 5–8), as well as the resulting floral differentiation (Figs. 9–10) are described. It was aimed at excluding the influence of plastochron changes on the interpretation of ontogeny of the shoot apex. For that reason two planes of longitudinal sections and two plastochron stages were compared. In young plants zonation does not become fully evident prior to floral differentiation. The anatomical structure of the shoot apex does not change substantially during the first two inductive cycles which proved to be obligatory under the given experimental conditions. The changes occurring during two further inductive cycles correspond to the total activation of the meristems as manifested by the growth and branching of the apex preceeding floral differentiation proper.     

Effect of (2-chloroethyl) trimethylammonium chloride (CCC) and gibberellic acid (GA3) on the flowering ofChenopodium rubrum L.

U rostliny krátkého dne Chenopodium rubrum L. se dá kvetení indukovat íty?mi 16hodinovými cykly tmy ji? 5 dní po vyklí?ení. Aplikace CCC v koncentraci 2.10^-3mvpr?běhu indukce kvetení zadr?uje a toté? piatí pro GA₃ i tehdy, je-li podáván ve velmi nízkých koncentracích (0,1 a? 0,01 mg/l). Av?ak sou?asná aplikace obou těchto látek v uvedených koncentracích vede k úplné reversi inhibice. Po p?enesení rostlin z média, obsahujícího CCC, na ?istý ?ivný roztok, inhibi?ní ú?inek CCC rychle zmizí a p?echází v slabou stimulaci. Na?e výsledky ukazují, ?e giberelin se zú?astňuje proces? kvetení i u rostliny krátkého dne.