The rate of RNA synthesis in shoot apices of the short-day (SD) plantChenopodium rubrum was compared in three successive dark periods required for flowering.³²P label was used for fractionation of RNA on slabs of polyacrylamide gels on mioroscopic slides. Incorporation of³²P and³H-uridine into apices was followed using histoautoradiography under identical conditions for oomparison. The lowest rate of synthesis was found on the seoond day of induction. A slight increase was usually observed in the third dark period, possibly linked with the first anatomical and/or morphological changes whioh have appeared due to mduction. Most of the label was found in ribosomal BNA in this case. After the plants wero transferred from light to darkness RNA synthesis in the shoot apex decreased within three hours. There was good agreement between results obtained by eleotrophoresis and by histoautoradiography. All previous observations which have been obtained using cytophotometry and histoautoradiography were confirmed.
The rate of synthesis of BNA in shoot apices is paralleled by that in primordial leavea. The marked rise of RNA synthesis during the inductive period, found in some planta requiring only single inductive cycle, was not established inChenopodium. A different pattern of RNA synthesis between plants flowering after one and after several dark periods, is suggested in discussion.

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.

The cells of some pea, tobacco and haplopappus callus strains reveal considerable variability in nuclear morphology (polymorphous nuclei, differences in nucleolar size and number, enlarged chromocentres) and chromosome counts. The specific features in the nuclear morphology of callus cells are related with some pecularities in the reproduction activity of these cells (amplification, amitosis, fragmentation, various deviations from normal mitosis) under cultural conditions including both the definite action of the culture system and the absence of the regulatory control by the intact organism.

The effects of various ammonium salts and amino acids on nitrite reductase (NIR) induction in isolated pea roots cultured in media containing nitrate or nitrite and either exogenous sucrose or no sugar were investigated. Thg aim of these investigations was to determine if the NIR level is subject to end-product control. The results showed that even though some ammonium salts and casamino acids can depress NIR level under certain conditions this inhibition cannot be interpreted in terms of direct end-product inhibition of NIR synthesis because their effects were dependent on the character (anion) and toxicity of the respective ammonium salt, on the presence of exogenous sucrose in the induction medium, and on the inducer of NIR. NH₄HCO₃ inhibited NIR induction at those concentrations which were toxic to the roots, ammonium phosphates hampered NIR induction only in roots exposed to nitrite in media containing sucrose, while casamino acids slightly depressed NIR induction only in roots exposed to nitrate and exogenous sucrose. The results further show that the basal (noninduced) NIR level changes little even under strongly toxic conditions.

Amaranthus viridis is a quantitative SD plant in which inflorescence primordia are initiated under both 24- and 8-h photoperiods after 12 and 10 days, when 8 and 7 leaves are differentiated, respectively. Photoperiod plays a non-determinate role, whereas the maturity of plants linked with the attainment of minimum leaf number is significant and of primary importance in floral induction. This is further confirmed by the more or less identical nature of changes in the total enzyme activity and isoenzyme patterns of peroxidase, esterase and alkaline and acid phosphatase under the two photoperiods. These changes occur once the minimum vegetative growth has been achieved prior to the reproductive transformation, irrespective of the photoperiod, pointing to the activation of a general common pathway of events leading to floral induction.

The increased activity of GOT (E.C.2.6.1.1.) and GPT (E.C.2.6.1.2.) transaminases in maize seedlings found as a marker of genotype opaque-2, was investigated in extirped sprouts of both genotypes, normal and opaque-2. The enzymatic activity was determined in three maize samples from breeding experiments, each sample consisting of a genotype pair, normal and opaque-2, collected from segregating ears of maize plants in the S₁ generation. The seedlings were aseptically grown for 7 days in two variants of cultivation, intact seedlings and sprouts extirped after 4 days of germination.
In the intact seedlings of genotype opaque-2 an increased activity of GOT and GPT, as compared to the intact normal plants, was observed. The extirpation of the sprouts enhanced GOT and GPT activity in the sprouts of both genotypes. However, in extirped sprouts the GOT and GPT activity was found to be still higher in the genotype opaque-2 as compared with the sprouts of normal genotype. Thus it seems that the increased transaminase activity in the sprouts of genotype opaque-2 is genetically determined. The increase does not result from an induction of enzyme synthesis through the supply of amino acids translocated from the endosperm to the sprouts. The absolute level of transaminases in the different breeding samples is dependent on the parenteral lines, the relative level of GOT and GPT activities is higher in the genotype opaque-2.

Growth correlations in the shoot apical meristem during transition to flowering were studied in a quantitative long day plant,Brassica campestris L. cv. Ceres, requiring only one long day for floral initiation. During photo-inductive exposure of the plants, an overall increase in cell number could be observed at the shoot apex concomitant with promotion of leaf initiation. Release from apical dominance and decline in relative growth rate of leaf primordia are reported as early effects of photo-induction. With the onset of floral differentiation, production of new leaf primordia had stopped altogether. Maximum increase in RNA concentration could be noticed in axillary meristems following photoperiodic treatment, whereas in vegetative plants the highest RNA concentration was found in leaf primordia. The significance of these changes occurring during transition to flowering is discussed.

In cotyledons ofChenopodium rubrum L. polydisperse RNA is synthesized in the region of the low molecular weight RNAs during photoperiodic induction. After short-time labelling the rate of 4s RNA synthesis was always higher in induced plants than in plants having obtained a light-break in the middle of the dark period. When glucose was added to the nutrient medium during the dark period of a single photoperiodic cycle the rate of nucleic acid (NA) synthesis was higher in non-induced plants than in induced ones at the termination of the dark period. In plants induced by two cycles in the absence of glucose the rate of NA synthesis at the termination of the second dark period was higher in induced than in non-induced plants. This difference is due to the differential kinetics of NA synthesis during darkness. In plants induced in the presence of glucose the peak of the rhythm in NA synthesis was advanced by 4 h relative to that found in plants induced in the absence of sugar. Thus, the termination of the dark period coincided with the negative slope of the oscillation in plants induced in the presence of glucose, while in plants having obtained a light-break NA synthesis decreased only slightly after having attained its peak. In plants induced in the absence of glucose the termination of the dark period coincided with the peak in the rhythm in NA synthesis. The rhythm in NA synthesis of the cotyledons during the dark period of an inductive cycle is out of phase with the rhythm in flower initiation.

The relationship between photoperiodically changed growth of leaves, cotyledons, hypocotyl, roots and flowering has been investigated inChenopodium rubrum. It was found that all the growth characteristics recorded in leaves and cotyledons,i.e. length, area, dry weight and chlorophyll content, were inhibited during three inductive photoperiods (16 h darkness, 8 h light-SD) as compared with control plants grown under continuous illumination. Similarly, the cessation of root elongation and a decrease in root dry weight were observed. On the contrary, the elongation and dry weight of hypocotyl are stimulated by SD. The degree of the effect exerted by SD on the growth of different organs depends both on actual growth stage and the number of SD photoperiods. Increased relative rate of growth of roots and cotyledons was recorded in plants transferred after SD treatment to continuous illumination. However, this rise possesses only transitional character and the relative growth rate of treated plants equals that of control ones afterwards. The above growth changes are discussed as a possible modifying factor of floral differentiation.

GA₃ as well as SA increase the protein content of the stem and the leaves at 1 day under both 8- and 24-h photoperiods. A new protein band with Rm 0.47 seems to be associated with floral bud initiation as it develops within 1-3 days in the stem as well as in the leaves of plants exposed to inductive treatments regardless of whether the induction is caused by 8-h photoperiods or by treatment with GA₃ of SA under 24-h photoperiods. Another band with Rm 0.23 developed only in the stem of water-as well as GA₃- or SA-treated plants under 8-h photoperiods. It may possibly be associated with extension growth.

The reversal of the cytogenetic effects of 5-fluordeoxyuridine, which are the gradual decrease of the mitotic index and the induction of chromosomal fragments was followed after addition of different concentrations of thymidine. Thymidine in the concentration 10⁴ M reversed both the effects of FUdR completely. The concentration of thymidine 10⁵ M partly reversed both the decrease of the mitotic index and the induction of chromosomal fragments. Uridine also caused some decrease of the cytogenetic effects of 5-fluordeoxyuridine.

The action of exogenously applied hormones in the induction of morphogenesis inLimnophila chinensis (Osb.) Merr. tissue culture has been demonstrated. Stem expiants were grown on Murashige and Skoog's medium containing various levels of kinetin, gibberellic acid and indole-3-acetic acid. Formation of roots, shoots (normal or abnormal), plantlets and friable, hard or nodulated calluses depended largely on the hormone levels used. The formation of normal shoots and roots were stimulated by treatment with kinetin. GA₃ treatment stimulated the bud differentiation but inhibited the root initiation. A combination of kinetin and GA₃ gave variable results.

Treflan has the ability to induce polyploidy inAllium cepa root tips. The frequency of polyploidy was reduced after allowing the roots to recover, which indicates that the process of polyploidy is a reversible one, if we apply Treflan for a short time (4 h). As soon as the chemioal is removed from the cells, they go on with their normal way in division. It was found that the concentration 350 × 10^-5 ml Treflan per 100 ml water is an effective concentration in producing polyploidy in the roots ofAllium cepa.
Treflan induced C-tumours in root tips ofAllium cepa. Two types of enlargement were noticed, complete swelling along the majority of root tips and complete swelling with a certain enlargement immediately after 2 mm terminal tips. The histological examination of these zones revealed that swellings were due to cell enlargement both in meristematic and in tumour zones. This conclusion was evidenced by the results obtained from the nucleoplasmic index.

Anatomical-histological investigations of slide series revealed that the secondary vascular formations appearing scattered in callus tissue culture derived from a stem piece containing mycoplasma-like organisms (MLO) (potato witches' broom agent) have no connection with the vascular tissues of the original stem piece. 77% of reconstituted tobacco plants (Nicotiana glauca Grah.) from these callus cultures were infected with MLO. When diseasedNicotiana glauca scions are grafted on healthy tomato plants as stocks, MLO transmigration into the stocks is realized in all cases ten days after grafting; at that time, the scions and stocks are united only by a zone of a few layers of parenchymatous cells without any indication of vascular tissue union between the two components. In both cases, the MLO transmigration could not be realized by passage through sieve pores in sieve tubes. Our results bring indirect evidence of MLO transmigration through parenchymatous cells and thus support the hypothesis of MLO passage through the plasmodesmata.

A technique is described for the establishment of infectious axenic callus cultures originated from stems of tobacco plants suffering from a mycoplasma disease, potato witches' broom. When stem pieces from a diseased tobacco plant (Nicotiana glaucaGrah.) are culture on a modified nutrient medium according toMurashige andSkoog (1962) with added kinetin (0.64 mg or 2.56 mg 1 000 ml^-1) and IAA (2 mg or 4 mg 1 000 ml^-1), the pathogen persists and spreads in the newly formed callus tissue. The presence of the pathogen was proved by implantation of callus slices into stems of tomato plants which indicate the disease very reliably. Reconstituted tobacco plants too recovered the infectious agent of potato witches' broom; 27 plants from the 35 reconstituted plants were diseased. Similar results were obtained with Crimean yellows. Calli were cultured and subcultured from 1 to 18 monthsin vitro and some infectious cultures are maintained and their investigation goes on. We suppose that kinetin plays a very important role for infectivity preservation in callus cultures.

The cytokinin-autonomous strain (A_s) of tobacco callus differs from the original cytokinin-dependent strain (D) and from the cytokinin- and auxin-autonomous strain (A₄) by a significantly lower activity of glucose-6-phosphate dehydrogenase (G-6-PDH). Changes in the total G-6-PDH activity were associated with differences in the number of G-6-PDH isozymes. The A_s strain contained only one isozyme, four isozymes were found in D and A₄ strains.

Barley embryos were treated by 0.1% colchicine for 30 min. Samples of root tips were fixed after 4 h, 8 h and 12 h. In the first sample,c-metaphases, normal metaphases and anaphases were present jointly. Inc-metaphases, chromosomes sometimes tended to make two groups with 7 chromosomes in each. In anaphases, lagging chromosomes, tripolar and multipolar anaphasos were found. No chromosomal aberrations were detected in anaphases and metaphases. No chromosome disturbances were found in root tips sampled 8 h and 12 h after colchicine treatment.

The participation of the genotype and of organ specifity effect in the quality of morphogenetic response (callogenesis, bud and root formation) of primary maize explants has been investigated. The presence of synthetic auxins - especially 2,4-D at 1 to 5 mg 1^-1 conc. - in cultivation medium was essential for both callus formation and continuous growth of tissue and suspension cultures. Anatomic structure of callus cultures is permanently heterogeneous, their growth is ensured by the action of meristems of the type found in root tips, and by repeated callogenesis from malformed roots. Adventive buds and plants could be regenerated only from cultures of embryonal origin (of one line). The presence or absence of the endosperm gene "opaque" did not influence callogenesis intensity in cultures of isolated embryos; however the morphogenetic response was clearly "line specific".

The three plant types ofAmaranthus namely,A. caudatus f.albiflorus, A. caudatus f.caudatus andA. tricolor var.tristis are qualitative short day plants with critical photoperiods 16.0, 15.5 and 15.0 h, respectively. Gibberellins A₃, A₄₊₇ and A₁₃ affect extension growth, leaf differentiation and floral induction differently. Thus, in all the three plant types ofAmaranthus, whereas, GA₃ and G₄₊₇ enhanced extension growth, GA₁₃ was completely ineffective under both, 24- and 8-h photoperiods. None of the three gibberellins could affect the leaf differentiation. In all the three plant types, flowering was promoted by GA₁₃ and not by other gibberellins tried. GA₁₃ caused promotion was manifested in two manners, firstly by lowering the critical dark period requirement in each inductive cycle, and secondly by shortening the total period taken for the initiation of inflorescence primordia under inductive photoperiods. The floral induction by gibberellins inAmaranthus is contrary to the gibberellin-anthesin concept of Chailakhyan. It is suggested that gibberellins other than GA₃ may be playing an important role in floral morphogenesis of short day plants.

Formation of stems both in callus tissue isolated from hypocotyl and in apical meristems culture of carnation plants (Dianthus caryophyllus L. ev. "Grenadin white, yellow, scarlet red, dark red and pink") was evokedin vitro using chemically defined medium. The rooted stems were transferred into pots and cultivated under natural conditions.

It is wellknown that following the amputation, or darkening of one cotyledon in decapitated flax seedlings, the opposite remaining, or illuminated, cotyledon exerts a stimulatory effect on the growth of its axillary bud. For the induction of this stimulating effect a 21-72 h continuous darkening of the cotyledon is sufficient. Endogenous gibberellins take part in the stimulation effect of the illuminated cotyledon, since their level in the illuminated cotyledon increases as early as 12-48 h following the darkening of the opposite cotyledon. The apical part of the cotyledon has a higher growth stimulatory effect on the growth of the cotyledonary axillary bud than the basal half. This again is associated with endogenous gibberellins the level of which is higher in the apical half of the cotyledon than in the basal one.
Upon removal of the root and hypocotyl base in decapitated flax seedlings deprived of one cotyledon, the remaining cotyledon loses its stimulatory influence, so that the bud of the amputated cotyledon grows more vigorously (Dostál 1955). In this growth correlative phenomenon the root may be substituted by cytokinin BA applied in the form of a 0.1-1.0 per cent paste onto the remaining cotyledon, for again in this case the bud of the preserved cotyledon grows more vigorously. Following the decapitation of the axillary of the amputated cotyledon in decapitated pea seedlings with an intact root and deprived of one cotyledon, the axillary of the remaining cotyledon grows more intensively than the serial of the removed one. If the plants operated on in the same way are deprived of the root, the serial of the removed cotyledon gains a correlative growth predominance. If the plants deprived of root are cultivated at the same time in a solution of BA (10-20 mg 1^-1), the correlative predominance is acquired by the axillary of the remaining cotyledon. In growth correlations between cotyledons and their axillary buds in pea seedlings the root may thus be substituted by exogenous cytokinin, as well.

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.

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.

The effect was studied of chloride ions, added in the form of different salts, on nitrate reductase (NR) level in excised pea roots, on anaerobic nitrite production in an assay medium lacking both nitrate and n-propanol, on nitrate content in the roots, and on in vivo NR activity determined in an assay medium containing 5% n-propanol. The presence of Cl in nitrate containing nutrient solutions resulted in lower NR levels, however counterions supplied together with Cl tended to modify slightly this general trend. The negative effect of Cl ions was also apparent, when Cl ions were applied before nitrate ions. Anaerobic nitrite production in the medium lacking both nitrate and n-propanol was not influenced by chloride ions. Nitrate content in the roots was reduced in the presence of chloride both at 3 mM and 15 mM NO₃ in nutrient solutions; however, at 16 mM NO₃, nitrate content in the roots exoeeded even in the presence of 15 mM Cl nitrate content in those root segments which were cultivated in a nutrient solution with 6 mM nitrate, which is the concentration at which NR reaches the level of saturation in excised pea roots. The results obtained suggest that a special induction nitrate pool exists in plant cells besides the storage and metabolic nitrate pools.

A study was made on certain problems connected with the application of methods of stigmatal and placental pollinationin vitro in the snapdragon.
Germinable pollen without microbial contamination could be obtained only from the flowers which were left, after surface sterilization, on the stem till the dehiscence of anthers. The germination of pollen, and especially the growth of pollen tubes was better in a 0.3 M lactose solution with 10^-3 per cent H₃BO₃ than in usual sucrose media.
The seedsin vitro could be obtained only after normal pollination and the successive artificial cultivation of the entire pistil (the method of stigmatal pollinationin vitro). The seeds did not differ either in their morphology or in their size from those developed under natural conditions, they were viable and without dormancy.
Though the pollinated pistils were cultivated in the medium without growth substances, callus formation from uncovered ovules and placenta was observed in some cases. A dependence was revealed of the proliferation on pollination and on the cultivar employed.
When using the method of placental pollinationin vitro we failed to obtain seeds. The main difficulty in the use of this method seems to consist in the fact that pollen tubes are not capable of growing from the surface of ovules and placenta into the micropyle.

Beginning with the second inductive cycle the rate of nucleic acid (NA) synthesis in cotyledons and apical buds ofChenopodium rubrum is higher at the end of the dark period or 4h following transfer of the plants to light in induced plants than in non-induced ones. This is due to an increase in all NA fractions. The greatest difference between NA synthesis in induced and non-induced plants was observed at the end of the second (or sometimes third) inductivecycle. In the subsequent cycles the difference decreased or disappeared eventually. During photoperiodic induction NA synthesis shows a diurnal rhythm with a peak at the end of the dark and at the beginning of the light period. Rhythmicity of NA synthesis is endogenous. The period length of the endogenous oscillation is about 18 h. Interruption of the dark period by light causea amplitude of the first oscillation to be reduced and delays the appearance of the second peak. NA synthesis did not show rhythmicity in plants grown in continuous light. The significance of the observed phenomena for photoperiodic induction is being discussed.

In this report, B₉ treatment had no effect on the growing of rosette biennialScrophularia vernalis L.; it inhibited or slowed stem elongation. Applications of GA₃ to B₉ treated plants produced a significant increase of stem elongation, in relation to GA₃ treated plants. Plants treated with only GA3 failed to flower; otherwise, the flowering of vernalized plants was not altered by GA₃. Thus, B₉ effect on flowering was tested by using GA₃. B₉ by itself induced flowering, it increased inflorescence formation in vernalized plants without altering stem growth pattern in the most of cases. The induction or the stimulation of flowering brought about by B₉ was not reversed by GA₃; we may thus hypothesize that flowering by B₉ oannot be traced back to gibberellin biosynthesis.

D-Glucose and several other sugars are taken up by slices of callus tissue ofNicotiana tabacum and are both oxidized to CO₂ and incorporated into tissue components. The uptake as well as the subsequent metabolism are slow, the transport process being characterized by lack of specificity, no hyperbolic saturation, no influence of metabolic or transport inhibitors and hardly any dependence on pH. The apparent activation energy of the process was about 42 kJ mol^-1. It appears that the transport proceeds by simple diffusion through narrow intercellular spaces and possibly hydrophilic pores and that only about 10% of the tissue volume is accessible to the sugars in question.

In two branched plants ofImpatiens balsamina with intact apex and leaves floral buds are induced only in the branch which is either exposed to 8-h (inductive) photoperiods or receives GA₃ treatment if maintained under 24-h (non-inductive) photoperiods. GA₃ induces floral buds on the treated branch even if the leaves on that branch are removed, showing that while leaves are essential for photoperception, these are not neoessary for GA₃ to cause induction. The effect of the inductive photoperiods or GA₃ treatments to a branch is not transmitted to the other branch which is treated with water and is maintained under non-inductive photoperiods even when the latter is defoliated but is transmitted if the apioal or both the apical and axillary buds on the branch receiving inductive photoperiods or GA₃ treatment are excised. It, therefore, appears that the existence of strong sinks in the form of axillary and apical buds on the treated branch prevents the transmission of photoperiodic as well as GA₃ effects to the other branch in this plant.

Tissue culture was isolated from the stem ofPopulus alba L. 'pyramidalis'. Callus formation was observed since November till March (1974),i.e. till the formation of calluses suitable for further subeultivation. The most vigorous growth was obtained with the callus culture cultivated on the nutrient medium of DIAZ-COLONet al. (1972) on which more than 11 g of fresh matter was produced after 30 d at the end of the first year of cultivation in darkness, with inoculum weight 1.5-1.8 g. A mild decrease in growth rate of the tissue culture was observed after the first year of cultivation. When illuminated, the originally yellow calluses turned green. The morphological and anatomical structure of the callus culture is also described and cell shape and cell size evaluated.