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.     

Endogenous auxins in apical buds ofChenopodium rubrum L. after application of (2-chloroethyl) trimethylammonium chloride (CCC)

Apical buds ofChenopodium rubrum from plants treated with CCC contain more endogenous auxins than buds from control plants, the level of these compounds increasing with the application of rising concentrations of the retardant. An especially marked increase was observed in the level of substance “X” which on chromatographic separation runs in the zone of tryptamine or its derivative. Since it has been shown in previous experiments that the inhibitory effect of CCC on flowering ofChenopodium rubrum may be reversed by indole-3-acetic acid (IAA) it is believed that the increase in auxins after application of CCC does not concern biologically active substances immediately available to the plant. It seems more likely that inactive precursors are involved which cannot be converted to the active substance in the presence of CCC, possibly due to blocking of the pertinent enzyme. If we assume that the wheat coleoptile used in the auxin bioassay in our experiments contains the pertinent enzyme it might convert the inactive precursors to active substances and, therefore, exhibit a growth stimulation even though the substances concerned would not necessarily be active in the buds from which they were extracted.     

The control of flower initiation by gibberellin in Helianthus annuus L. (sunflower), a non-photoperiodic plant

The involvement of gibberellins in the control of flowering of sunflower was studied by direct application of GA₃ to the apex of the plants, analysis of the endogenous levels of gibberellin-like substances at different plant ages, and indirectly by the application of paclobutrazol, an inhibitor of gibberellin synthesis. GA₃ speeded-up flower initiation and floral apex development. The time of GA₃ application was more critical than the amount of GA₃ applied. The endogenous levels of gibberellin-like compounds increased significantly by day 15 after sowing. The application of paclobutrazol markedly delayed floral initiation and this effect was also depedent on plant age. Both GA₃ and paclobutrazol had their greatest effects between 10 and 20 days after sowing suggesting that an increase in gibberellins in that time period plays a role in floral initiation.     

Gibberellins in the control of photoperiodic flower transition in Pharbitis nil

The role of gibberellins in the photoperiodic flower induction of short-day plant Pharbitis nil has been investigated. It has been found that the endogenous content of gibberellins in the cotyledons of P. nil is low before and after a 16-h-long inductive dark period. During the inductive night the content of gibberellins is high at the beginning of darkness and about the middle of the dark period. Exogenous GA₃ when applied to the cotyledons of non-induced plants does not replace the effect of the inductive night but it can stimulate the intensity of flowering in plants cultivated on suboptimal photoperiods. GA₃ could also reverse the inhibitory effect of end-of-day far-red light irradiation on P. nil flowering. 2-Chloroethyltri-methylammonium chloride (CCC) applied to the cotyledons during the inductive night also inhibited flowering. GA₃ could reverse the inhibitory effect of CCC. The obtained results strongly suggest that gibberellins are involved in the phytochrome controlled transition of P. nil to flowering. Their effect could be additive to that of photoperiodic induction.     

Indole auxins in spinach plants grown in long and short days

The endogenous auxin-like substances were analyzed in the shoot extracts of young spinach seedlings, exposed to photoperiodic induction. At least eight indole auxins were found. One of them was identified as tryptophan, the other one is most probably IAA. The plants grown in long days had a higher level of ether soluble auxins than the controls in short days. Separate extractions of plants after each of the eight inductive days showed that the auxin content was not constant, but subjected to irregular oscillations. However, parallel oscillations were also found in control plants grown in short days. Staminate plants were found to contain more endogenous auxins than the pistillate ones. It is concluded that the quantitative changes in auxins during the photoperiodic induction are probably not related to flowering, but to some other growth process, common to all plants in that phase of growth. The higher level of auxins in staminate plants may be the cause of their faster elongation before the onset of flowering.     

Effects of exogenous cytokinins on flowering of the short-day plantChenopodium rubrum L.

Kinetin at a concentration from 3.10^-6 M to 1.10^-3 M was applied to the plumule ofChenopodium rubrum plants during photoperiodic induction. Different levels of induction were compared (one and three short days). The higher concentrations of kinetin applied to induced plants inhibited flower formation. The rate of leaf initiation was increased under these treatments. Lower concentrations of kinetin (from 3.10^-6 M to 1.10^-5 M) usually promoted lateral bud formation and flowering. The step-wise application of kinetin revealed that the inhibitory effect on flowering had been restricted to the inductive period. The effects of kinetin, benzyladenine and trans-zeatin were compared in plants partially induced by two short days. High concentrations always inhibited flowering. Benzyladenine was the most effective in this respect. Root removal diminished the inhibitory effects of cytokinins on flowering as was stated with benzyladenine. It is assumed that endogenous cytokinins play a role in the regulation of organogenetic activity of the stem apical meristem. Depending on the photoperiodic conditions, they presumably exert their activity by maintaining the vegetative functions of the apex.     

Action du CCC et du B 995 sur la mise à fleur d'une plante bisannuelle,l'Œnothera biennis

Summary Oenothera biennis L. is a typical biennial plant with an absolute cold requirement for subsequent floral initiation under long-day conditions. Flowering of vernalized Oe. biennis is associated with transition from a rosette habit to the formation of a long flower-bearing stem. Vernalization in Oe. biennis consists of two consecutive stages: (1) preparation for flowering; (2) preparation of stem elongation. At the end of the second stage, the level of endogenous gibberellin-like substances is sufficient to allow the stem elongation which is requisite for subsequent floral development.Applications of (2-chloroethyl)trimethylammonium chloride (CCC) via the roots, when made after the cold treatment, have no effect on internodal elongation and subsequent flowering of vernalized Oe. biennis. Relatively small amounts of CCC applied onto the apical bud of the rosette promote stem growth. However, 750 g of CCC act as a growth inhibitor and delay the beginning of stem elongation but this retardant has no effect on the number of flower buds.In contrast, N-dimethylaminosuccinamic acid (B 995), when applied after a cold treatment, delays the beginning of stem growth, this delay increasing with greater amounts of applied B 995. However, with relatively small amounts of B 995, the rate of later stem growth increases rapidly so that treated plants flower at the same time as controls.If gibberellic acid (GA₃) is applied on the apical bud just at the same time as B 995, it reverses the effects of the growth retardant, even when the amount of B 995, applied by itself, is sufficient to inhibit entirely stem elongation and flowering.B 995 is particularly effective if the preceding cold treatment was just sufficient for effective vernalization. If the cold treatment was extended beyond this time, a greater amount of retardant was required to obtain the same degree of stem growth inhibition.B 995 probably acts by interference with the metabolism of endogenous gibberellin-like substances. The delay of floral initiation in vernalized Oe. biennis by B 995 is a consequence of the retarded internode elongation; if this retardation exceeds a certain limit, the plant is devernalized and exhibits a cold requirement identical with the original one.     

Gibberellins are not essential for photoperiodic flower induction of Pharbitis nil

The influence on photoperiodic flowering of (2-chloroethyl)trimethylmmonium chloride (CCC), an inhibitor of gibberellin (GA) biosynthesis, was studied in the short-day plant Pharbitis nil cv. Violet. The cotyledons contained high levels of endogenous bioactive gibberellins, whereas in the plumules and first leaves the levels were low or undetectable. The first leaf responded to a single'dark treatment by inducing flowering when it was 10 mm or wider. Similar seedlings, but without cotyledons, were used as the assay plants to study the effect of CCC on photoperiodic flowering. Treatment with CCC had no effect on flowering of seedlings without cotyledons, although stem elongation was inhibited. By contrast. CCC inhibited flowering of the intact seedlings with cotyledons. Gibberellic acid applied to the shoot apex or to the first leaf promoted flowering in the CCC-treated seedlings without cotyledons. The results indicate thai gibberellins are not essential for the flower induction process in leaves, but that they promote flower initiation and/or later processes in the shoot apices.     

On the interaction of growth retardants with IAA and kinetin

In the pea test a highly positive response to the treatment with IAA reversed to a negative one or became 5 to 6 times weaker when CCC was applied together with IAA. In cultivating pea seedlings, following their decapitation, for two days in a 0.25 per cent CCC solution and then in water, growth of their cotyledonous axillaries (cotylaries) were inhibited. This inhibitive action of CCC could be made ineffective when the seedlings, following two-days’ cultivation in the CCC solution, were grown further in kinetin solutions (0.37–3 mg per 1). Cotylaries of decapitated pea seedlings, when grown in kinetin solutions were inhibited. With kinetin solutions of 6–12 mg/l a strong inhibition also occured in the growth of roots at the apical parts of which spherical swellings were developing. The CCC supplied to the roots of intact etiolated pea seedlings is translocated acropetally into the stem at a rate of about 5 cm per hour. Decapitation of the plant causes retardation of this transport, yet a coat of 0.00001–1% IAA or kinetin paste produces acceleration of the stream. Existence of an antagonism between CCC and IAA, demonstrated earlier, was found holding true also for B-9 (N, N-dimethyl-aminesuccinamic acid) and IAA, as the inhibitive action of B-9, 0.06% solution on the growth of lettuce hypocotyls was reduced to a highly significant degree when the plants were supplied with B-9 together with IAA at a concentration of 10 mg/l.     

光皮树花芽分化过程中内源激素含量变化的研究

以4年生光皮树嫁接苗为材料,采用酶联免疫吸附法（ELISA）测定同龄开花光皮树与不开花光皮树叶内4种内源激素生长素（IAA）、赤霉素（GA3）、玉米核苷素（ZR）、脱落酸（ABA）含量的动态变化,研究成花过程中叶片内源激素含量的变化与成花的关系。研究结果表明：在芽分化时期开花光皮树和未开花光皮树叶内4种内源激素含量动态变化存在显著差异。有花芽分化的光皮树保持相对较低和稳定IAA、GA3含量,ABA含量高且变化幅度较大,ZR含量相对较低,但随着花的形成含量逐渐升高;而在相同的生长季内,无花光皮树IAA、GA3含量先升高后降低,ABA含量先逐渐升高,然后下降,而ZR含量呈现出先逐渐升高然后降低,再升高再下降的变化趋势。因此,4种内源激素含量的动态变化影响光皮树的成花过程。     

Enhancement of lateral bud growth in Coleus blumei BENTH. by (2-chloroethyl) trimethylammonium chloride (CCC)

The relationship of GA to apical dominance in Coleus was examinedby substituting 1 % IAA, in lanolin, for the shoot apex of CCC-treated,control and GA-treated plants containing, theoretically, hyponormal,normal and hypernormal GA levels, respectively. The greatestinhibition of lateral bud growth was obtained in the treatmentcombining 1 % IAA and 100 ppm GA, suggesting that GA may beimportant in the apical dominance of Coleus. CCC inhibited main axis growth, reduced the level of endogenousGA and caused a marked release of lateral buds from apical dominance. The significant stimulation of lateral bud growth by CCC couldnot be ascribed to reduced endogenous GA since it was not reversedby exogenous GA, or by GA plus IAA, whereas 100 ppm GA overcamethe inhibition of main axis growth by CCC. It was also shownthat the CCC stimulation was not a result of compensatory growth,that is, enhanced lateral bud growth resulting from reducedapical bud growth. The CCC effect on lateral buds was interpretedas involving a system independent of auxin and GA or else apossible immobilization of auxin in addition to inhibition ofGA biosynthesis. (Received December 5, 1967; )     

多效唑调节水稻植株生长的作用机理

多效唑处理水稻植株后使株高降低和分蘖增加,植株体内的GA类物质和IAA含量相应下降,乙烯释放率显著增加;用外源GA_3和氮肥能逆转多效唑的抑制作用而使株高增加,同时GA类物质含量也明显增加。因此,多效唑不仅调节体内的GA类物质含量,还能通过调节IAA含量和乙烯释放率而影响植物生长。     

Effects of uniconazole and GA3 on cold-induced stem elongation and flowering of Raphanus sativus L.

The effects of gibberellin (GA) on cold-induced stem elongation and flowering of Japanese radish (Raphanus sativus L.) were investigated using application of GA3 and a GA-biosynthesis inhibitor, uniconazole (UCZ). UCZ very strongly inhibited stem elongation and delayed flowering, and the inhibition and delay were completely reversed by GA3 application. These results suggest that GA is necessary not only in the stem elongation but also in the flowering. When cold treatment (CT) was conducted on the plants whose GA level was lowered by UCZ, GA3 applied after CT completely reversed the delay of flowering. Thus low GA level probably did not retard cold induction. Microscopic observation of apical meristem showed that UCZ delayed flowering by delaying the shift from vegetative to dome-shaped meristem. This result suggested that low GA level delayed floral evocation. Consequently it was suggested that low GA level retarded physiological process involved in long day induction or in floral evocation, resulting in delay of floral evocation.     

Exogenous regulators in the mycelium ofPleurotus ostreatus after exogenous application

The mycelium of the fungusPleurotus ostreatus was cultured on a synthetic medium with growth regulators. Best growth was observed on media with 100 ppm IAA, 200 ppm GA₃ and 200 ppm of kinetin. Growth of terminal hyphae and the appearance of colonies differed characteristically. Auxin bio-assays showed that the content of substances in younger mycelium slowly increased during cultivation without conspicuous changes in the medium. After exogenous application of IAA the content of auxins in younger mycelium increased and the content of IAA decreased gradually in the culture medium. Gibberellin bio-assays showed their content in the mycelium fell during cultivation and while it increased in the medium. Application of GA₃ to the medium leads to a significant increase in these substances in the young mycelium and in the medium their level remains the same. Cytokinin bio-assays showed their content to vary during cultivation. An addition of kinetin to the nutrient medium results in an increase of cytokinins in the mycelium while in the medium the level of cytokinins is maintained. Addition of one substance thus influences the levels of other endogenous regulators.     

Action of inhibition by fluorodeoxyuridine and its reversal by thymidine on the nucleic acid fractions of the overground parts of photoperiodically inducedChenopodium rubrum plants

The nucleic acid fractions obtained by chromatography on MAK columns were compared in 4 variants ofChenopodium rubrum plants treated in different ways during floral induction. The first variant was normally induced to flowering. The second one was inhibited by application of FUDR to the apical bud on the third day of induction. In the third variant the inhibitory effect of FUDR was reversed by application of THY 24 h after FUDR treatment. In the fourth variant THY was applied 24 h after the termination of induction at a time when it was no longer able to reverse the inhibition of flowering. In plants treated with FUDR, a decrease in DNA and RNA synthesis was observed. After reversal of the inhibitory effect of FUDR by THY, DNA synthesis remained somewhat lower than in the control variant but RNA synthesis reached the same level as in the controls or even surpassed it. In plants to which THY was applied at a time when it was no longer possible to reverse flowering, the results obtained from different experiments were not identical. In some experiments nucleic acid synthesis remained lower than in the variant in which THY restituted flowering. In other experiments nucleic acid synthesis was fully restituted and reached the same level as in the control variant. This phenomenon is explained by the different size of the experimental plants at the beginning of the different experiments. The results are discussed with respect to the relation of the dynamics of nucleic acids to growth and development of the plants.     

Gibberellins and the floral transition in Sinapis alba

The putative role of gibberellins in the transition to flowering was investigated in Sinapis alba , a caulescent long-day (LD) plant. It was observed that: (1) physiological doses of exogenous gibberellins (GA₁, GA₃, GA₉) do not cause the floral shift of the meristem when applied to plants grown in short days but have some positive effect on the flowering response to a suboptimal LD; no inhibition was observed in any case; (2) GA-biosynthesis inhibitors (prohexadione-Ca and paclobutrazol) considerably inhibit stem growth but have some negative effect on flowering only when a suboptimal LD is given; and (3) the floral transition induced by one 22-h LD does not correlate with any detectable change in GA content of the apical bud, of the leaves, and of the phloem exudate reaching the apex. Taken together, these results suggest that GAs do not act as a major signal for photoperiodic flower induction in Sinapis .     

The Effect of Growth Substances on Growth of Excised Embryo Shoot Apices of Wheat in Vitro

Excised embryo shoot apices of wheat were grown on media containingeither indolylacetic acid (IAA), kinetin, gibberellic acid (GA),or adenine. GA and adenine stimulate the activity of the apicalmeristem whereas IAA and kinetin do not. No combination of anytwo substances significantly stimulated growth when comparedwith the most active substance present. GA can overcome theinhibitory effect of IAA or kinetin. The possible function ofthese substances in the apical meristem is discussed.     

Changes in the content of endogenous auxins in apical buds ofchenopodium rubrum L. Induced with respect to the endogenous rhythm in capacity to flower

The content of endogenous auxins was examined in apical buds ofChenopodium rubrum plants induced by a photoperiodic cycle of 16h darkness and 8h light followed by a dark period of various duration so as to correspond with either maximal or minimal flowering response in the endogenous rhythm in capacity to flower initiated by the photoperiodic treatment. Apical buds of potentially generative plants contained less auxins than apical buds of plants which remained in the vegetative state. Apical buds from plants treated with kinetin (1. 10^-3 M) and therefore remaining in the vegetative state showed an auxin level comparable to that of untreated plants exhibiting minimal flowering response irrespective of the duration of the second dark period. Plants cultivated on a sucrose solution (0.6 M) during the second dark period became generative even at the normal minimum of flowering. The auxin content of the apical buds was low, similarly as in untreated plants induced for a period leading to maximal flowering response. On the other hand, apical buds from plants grown on sucrose solution during a dark period leading to the manifestation of maximal flowering response showed a relatively high auxin content comparable to that found in untreated plants which had obtained a more extended induction by three photoperiodic cycles. The results are discussed with respect to the possible role of endogenous auxins in the regulation of the changes in growth correlations occurring in the shoot apex during photoperiodic induction and in the expression of the competence to flower.     

The effect of plant growth regulators and their interaction with electric current on winter wheat development

The aim of the experiments presented here was twofold. On the one hand, to study the influence of plant growth regulators, i.e. auxins (IAA and NAA), cytokinins (kinetin, zeatin and zeatin riboside), gibberellins: (GA₃) and polyamines (spermidine, spermine and putrescine) on the generative development of winter wheat plants, while on the other to study the interaction of plant growth regulators with electric current in affecting the efficiency of the flowering of wheat plants. Winter wheat var. Grana was subjected to a short vernalization (2 weeks at 5°C + 10 days at 10°C) on Murashige and Skoog medium containing growth regulators at different concentrations. For selected substances, seedlings were additionally treated with electric current by applying a constant voltage for a given period of time (30 V for 30 s or 1.5 V for 1 h) with the anode or the cathode inserted into apical leaves and the reference electrode into the media. After that, the seedlings were transferred to a glasshouse (20/17°C) where they were grown until heading was achieved. All the substances studied stimulated the generative development of winter wheat, in contrast to non-treated plants (control), especially at their highest concentrations. Spermidine, kinetin and GA₃ were the most effective in this process as they stimulated 100–80% of generative plants (whereas for the control the percentage stood at only 30% plants). The studied regulators also increased the rate of generative development, i.e. shortening the length of the vegetative phase (by about 30% in comparison with the control). Moreover, these substances treatments decreased the fresh mass of both seedlings and flowering plants and increased the number of spikelets in the ear. The electric current treatment interacted with the applied substances: the anode insertion into the leaves generally increased, whereas the cathode insertion decreased, the inductive effect of the growth regulators on generative development. However, the inhibition of the flowering process was more pronounced than the induction. Moreover, the passage of electric current shortened the time to heading. This decrease in the length of the vegetative phase was more visible after the anode was inserted into the leaves. The least noticeable effect of the electric current interaction with plant growth regulators on both the generative development and the time of heading was observed for auxins.