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.     

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.     

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.     

The rate of cell division in the shoot apical meristem during photoperiodic induction and transition to flowering

Cell division contributing to longitudinal growth of the shoot apex was investigated inChenopodium rubrum in segments marked by the axils of leaf primordia. Plants treated with two short days (16h of darkness and 8h of light) were compared with two non-induced controls (cultivated in continuous light or treated by alternations of 8 h of darkness and 4 h of light for two days). During the short-day treatments the rate of cell division contributing to the longitudinal growth decreases in all segments of the shoot apex irrespective of whether the darkness was given in inductive or non-inductive photoperiods. The rate of cell division contributing to longitudinal growth increases in the upper internodes of the shoot apex after the termination of the photoperiodic treatment and transfer of the plants to continuous light. However, cell division remains inhibited in the lowest segment of the shoot apex. This inhibition in the differentiating parts of the shoot apical meristem is a direct consequence of photoperiodic induction. It is supposed that this inhibition is related to evocation similarly as the well-known phenomenon of stimulation of cell division in the apical dome.     

HISTOCHEMICAL AND AUTORADIOGRAPHIC STUDIES OF FLORAL INDUCTION IN CHENOPODIUM ALBUM

Gifford , Ernest M., Jr. , and Herbert B. Tepper . (U. California, Davis.) Histochemical and autoradiographic studies of floral induction in Chenopodium album. Amer. Jour. Bot. 49(7): 706–714. Illus. 1962.—Chenopodium album was induced to flower using short-day photoperiods. Changes in the chondriome, starch, total protein, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), and histone distribution in cells of vegetative and inflorescence shoot apices were studied. The distal cells of the vegetative apex (especially the axial tunica cells) possess larger nucleoli and vacuoles, less granular mitochondria, and more differentiated plastids than do other cells of the apex; the distal cells stain lightly with dyes that indicate the presence of DNA and histone. RNA is distributed relatively uniformly in the shoot apex; the cells at sites of leaf initiation and young leaf primordia contain slightly higher concentrations of RNA than the axial cells of the shoot apex. Protein is uniformly distributed throughout the vegetative as well as the inflorescence apex. Upon induction, the chemical and morphological differences between cells in the shoot apex gradually disappear. RNA concentration of cells in the apex increases, reaching a maximum after 4 inductive cycles. Protein concentration of cells also increases, but this increase lags behind that of RNA.     

STEREOLOGICAL STUDY OF ULTRASTRUCTURAL CHANGES IN THE SHOOT APICAL MERISTEM OF NICOTIANA TABACUM DURING FLORAL INDUCTION

Shoot apices of a short-day sensitive line of Nicotiana tabacum L. cv. NCTG-22 have been examined by electron microscopy for ultrastructural changes which occurred in the central zone over a 17-day period during the transition from vegetative to reproductive growth. Plants were grown in controlled-environment chambers of the NCSU Phytotron and exposed to an inductive photoperiod after a 6-wk juvenile phase. Ultrastructural changes were investigated from photomicrographs using a semi-automatic stereological procedure and a microcomputer. After exposure to only one inductive cycle cell and nuclear cross sectional areas in short-day plants were significantly larger than in long-day controls. Subsequently, under short days cross sectional areas of cells, nuclei, vacuoles and proplastids decreased, while mitochondrial cross sectional area and relative volume increased during the course of the induction period. In induced apices as cross sectional areas and relative volumes of vacuoles and proplastids decreased, their profile numbers increased. The reduction in cross sectional areas of cells and most organelles was associated with an increase in rate of leaf initiation and size of the apical dome. The demand for sufficient energy input to maintain the surge in growth and activity preceding floral initiation was reflected by the significant increases in cross sectional area, profile numbers and density of the mitochondria population. Even though the transition period is quite long for Nicotiana, cells and organelles in the central zone were observed to progress through similar changes in morphology that are known to occur in Sinapis and Xanthium which exhibit a more rapid and absolute response to photo-induction.     

Floral determination in the terminal bud of the short-day plant Pharbitis nil

Temporal and spatial aspects of floral determination in seedling terminal buds of the qualitative short-day plant Pharbitis nil were examined using a grafting assay. Floral determination in the terminal buds of 6-day-old P. nil seedlings is rapid; by 9 hr after the end of a 14-hr inductive dark period more than 50% of the induced terminal buds grafted onto uninduced stock plants produced a full complement of flower buds. When grafted at early times after the end of the dark period the terminal buds of induced plants produced three discrete populations of plants: plants with no flowers, plants with two axillary flowers at nodes 3 and 4 and a vegetative terminal shoot apex, and plants with five to seven flowers including a terminal flower. The temporal relationship among these populations of plants produced by apices grafted at different times indicates that under our conditions, the region of the terminal bud that will form the axillary buds at nodes 3 and 4 becomes florally determined prior to floral determination of the region of the terminal bud giving rise to the nodes above node 4.     

Bi-directional inflorescence development inArabidopsis thaliana: Acropetal initiation of flowers and basipetal initiation of paraclades

In this study we investigated Arabidopsis thaliana (L.) Heynh. inflorescence development by characterizing morphological changes at the shoot apex during the transition to flowering. Sixteen-hour photoperiods were used to synchronously induce flowering in vegetative plants grown for 30 d in non-inductive 8-h photoperiods. During the first inductive cycle, the shoot apical meristem ceased producing leaf primordia and began to produce flower primordia. The differentiation of paraclades (axillary flowering shoots), however, did not occur until after the initiation of multiple flower primordia from the shoot apical meristem. Paraclades were produced by the basipetal activation of buds from the axils of leaf primordia which had been initiated prior to photoperiodic induction. Concurrent with the activation of paraclades was the partial suppression of paraclade-associated leaf primordia, which became bract leaves. The suppression of bract-leaf primordia and the abrupt initiation of flower primordia during the first inductive photoperiod is indicative of a single phase change during the transition to flowering in photoperiodically induced Arabidopsis. Morphogenetic changes characteristic of the transition to flowering in plants grown continuously in 16-h photoperiods were qualitatively equivalent to the changes observed in plants which were photoperiodically induced after 30 d. These results suggest that Arabidopsis has only two phases of development, a vegetative phase and a reproductive phase; and that the production of flower primordia, the differentiation of paraclades from the axils of pre-existing leaf primordia and the elongation of internodes all occur during the reproductive phase.     

Correlations between Growth and Flowering in Chenopodium amaranticolor: I. Initiation of Leaf and Bud Primordia

Vegetative plants of Chenopodium amaranticolor were inducedto flower by exposure to 2, 6 or continuous short days (SDs)and the effect of such treatments on organogenesis at the apexof the main stem followed by means of dissections. The mostoutstanding responses to SD treatment were (I) an immediateelongation of the apex, (2) a stimulation of the rate of initiationof leaf primordia, and (3) a promotion of the rate of initiationof axillary bud primordia. In response to as few as 2 SDs, therate of initiation of leaf primordia increased from 0.47 toa maximum of 3.70 per day and the rate of initiation of axillarybud primordia immediately increased from 0.47 to 1.35 per day. Precocious initiation of axillary bud primordia led to the formationof double ridges. The results indicate double ridges to be homologouswith vegetative axillary buds; although they normally developedinto reproductive tissues, they passed through a period of vegetativegrowth following minimal induction to flowering by exposureto 2 SDs. The rate and degree of flowering were highest in plants whichreceived the longest period of SDs, but the differences in finalflowering response were greater than the differences betweenthe initial responses at the apices. The effect of SDs was thusnot confined to an initial stimulation of organogenesis; a prolongedexposure to SDs must have enhanced the subsequent developmentof double ridges into flower primordia. The results are discussed in relation to previous findings andthe general conclusion drawn that the initiation of double ridgesis very widely accompanied by a stimulation of apical growth.It is suggested that inductive conditions remove a general growthinhibition and that the resultant stimulation of apical growthmight lead to the initiation of double ridges.     

The ABSCISIC ACID-INSENSITIVE 3 (ABI3) gene is expressed during vegetative quiescence processes in Arabidopsis

The ABSCISIC ACID-INSENSITIVE 3 ( ABI3 ) gene of Arabidopsis thaliana (L.) Heynh is known to play an important role during seed maturation and dormancy. Here, we present evidence suggesting an additional role for ABI3 during vegetative quiescence processes. During growth in the dark, ABI3 is expressed in the apex of the seedlings after cell division is arrested. The 2S seed storage protein gene, a target gene of ABI3 in seeds, is also induced in the arrested apex under similar darkness conditions. In addition, β -glucuronidase expression under the control of the ABI3 promoter is abolished by treatments that provoke leaf development in the dark [sucrose and abscisic acid (ABA) biosynthesis inhibitors] and induced by treatments that prevent leaf development (darkness and ABA). Furthermore, ABI3 expression is absent in apices of dark-grown de-etiolated ( det 1 ) and abi3 mutants, both known to develop leaves or leaf primordia in the dark. The fact that the expression of the ABI3 gene is only observed in a fraction of the analysed plants suggests that ABI3 is probably only one of the components of a molecular network underlying quiescence. In addition to the expression of ABI3 in apices of dark-grown seedlings, the ABI3 promoter confers expression in other vegetative organs as well, such as the stipules and the abscission zones of the siliques. In conclusion, apart from its role in seed development, ABI3 might have additional functions.     

The RING Finger Protein Nt RCP1 Is Involved in the Floral Transition in Tobacco(Nicotiana tabacum)

The transition from the vegetative phase to the reproductive phase is a major developmental process in flowering plants.The underlying mechanism controlling this cellular process remains a research focus in the field of plant molecular biology.In the present work,we identified a gene encoding the C3H2C3-type RING finger protein Nt RCP1 from tobacco BY-2 cells.Enzymatic analysis demonstrated that Nt RCP1 is a functional E3 ubiquitin ligase.In tobacco plants,expression level of Nt RCP1 was higher in the reproductive shoot apices than in the vegetative ones.Nt RCP1-overexpressing plants underwent a more rapid transition from the vegetative to the reproductive phase and flowered markedly earlier than the wild-type control.Histological analysis revealed that the shoot apical meristem of Nt RCP1-overexpressing plants initiated inflorescence primordia precociously compared to the wild-type plant due to accelerated cell division.Overexpression of Nt RCP1 in BY-2 suspension cells promoted cell division,which was a consequence of the shortened G2 phase in the cell cycle.Together,our data suggest that Nt RCP1 may act as a regulator of the phase transition,possibly through its role in cell cycle regulation,during vegetative/reproductive development in tobacco plant.     

Histochemical study of vegetative and floral bud formation in tobacco stem expiants

Primary explants from the inflorescence stem of tobacco and primary explants from the stem of vegetative plants, cultivatedin vitro under the same conditions, display different morphogenetic ability. The former give rise mostly to floral buds, whereas the latter exclusively to vegetative ones. Histological and histochemical analyses of both original andin vitro cultivated explants were made. They showed differences in chlorophyll content and alcohol dehydrogenase (AD) activity of the original explants reflecting their different metabolic status. Bud primordia were initiated in the superficial meristematic layer derived from epidermal tissues. Floral or prefloral apices were characterized by a strong AD activity in all cells of the meristem, while in vegetative apices AD activity was restricted to their uppermost parts. A high rate of procambium differentiation connected with leaf primordia formation was typical of vegetative buds. A higher concentration of glucose (5 %) enhanced cell division in explants, which is also correlated with a higher AD activity. The significance of vascular tissues for differentiation of vegetative buds is discussed. Presented at the International Symposium “Plant Growth Regulators” held on June 18-22, 1984 at Liblice, Czechoslovakia.     

The Structure of the Plumule of Nelumbo Nucifera Gaertn. and the Nature of its Scale

The structure of the plumule of Nelumbo nucifera Gaertn. and its feature covered with scale are seldom seen in dicotyledon. The fact that the plumule possesses scale is even more uncommon. This particular phenomenon is investigated by observing the differentiation of the plumule apex and the development of the leaf organs. After the seed is formed, the embryo has two young leaves and a terminal bud covered with scale. In the bud it has already differentiated the 3rd and the 4th leaf primordium and a shoot apex, the differentiation of which is very complex. So the structure of the plumule passes through 4 plastochrons altogether. It is made clear through observation and analysis that, before the 4th leaf primordium is formed, the transforma- tions of the shoot apex of the embryo in each plastochron are fundamentally alike. After the 4th leaf primordium is developed, the shoot apex becomes complex and there appear 3 different active cell regions which become the bases of vegetative bud of the seeding apex. The development of these 3 active cell regions will be stated in “The Structure of the Vegetative Bud of Nelumbo nucifera Gaertn. and the Nature of its Scales.” The apices of the plumule are almost slightly domed in structure. As a rule, their width is from 95 to 107 μ. Their height is from 17 to 20 μ during one plastochron. Before the 3rd leaf initiation, the anatomical structure of apices is examined and the fol- lowing zones may be delimited: zone of tunica initials, zone of corpus initials, peripheral zone, and zone of rib meristems. It is frequently observed that the cell of corpus in subapical peripheral zone develops periclinal division, which is the initial cell of leaf primordium; Procambium will appear before the stage of the appearance of leaf buttress. The apex of the plumule is in an apical position, but when the seedling is formed, as the developing leaves are alternate, the directions of the shoot apex are changed, simultaneously the base part of the leaf encloses the axis, and the adaxial meristem also differentiates the scale which encloses the terminal bud, thus placing the bud in axillary of the leaf and forming a zigzag phenomenon of the axis of the seedling. Above the basal adaxial side of the leaf primordium develops the scale of the plumule with meristem periclinal division of closely attached protoderm as its base. So the scale of the plumule of Nelumbo nucifera Gaertn. and the axillary stipule are of the same origin. To sum up, the scale of the embryo of Nelumbo nucifera Gaertn. is differentiated from the adaxial meristem of the basal part of the leaf primordium, and is the derivative part of the leaf. It has the same function as the coleoptile of the monocotyledon. Whether they are homologous organs or not is still to be investigated.     

Rates of cell division in the young prefloral shoot apex ofChrysanthemum segetum L.

Summary The rate of cell division was determined by the colchicine induced metaphase-accumulation technique in the young prefloral shoot apex of the quantitative long-day plantChrysanthemum segetum L. growing under conditions favourable to flowering (16-hour photoperiod; 124Em^–2s^–1; 22 °C). Cell cycle duration was evaluated in relation to the location of the cells in the intact apex. The cell cycle durations were 53.5 hours, 47.4 hours, and 97.7 hours in the axial, lateral and subapical central cells respectively. Compared with previous results, these data give evidence of the major role played by the early increase in cell division rate of axial cells in the new pattern of the prefloral shoot apex at its initial stage of development. By comparison with the vegetative shoot apex, the cell cycle duration was preferentially shortened in the axial zone; it was only slightly altered in the lateral zone while it was lengthened in the vacuolating subapical central cells. In the three zones within the prefloral shoot apex, the duration of mitosis was constant (3.2 to 3.3 hours) and the same as in the vegetative shoot apex.     

The anatomy of the shoot apex ofHyoscyamus niger L. reversed from the generative to the vegetative state

A study was made of the anatomical structure of the shoot apices ofHyoscyamus niger L. in plants which were transferred from a long-day to a short-day regime after the initiation of the inflorescence. After a certain time these plants are reverted to the vegetative stage with the inhibition of the development of further flower buds and the renewed production of rosette leaves. The inflorescence apex consisted of a few superficial layers of cells and a corpus composed of slightly elongated cells. The anatomical structure of the apices which were reverted into the vegetative state resembled that of shoot apices in the intermediate stage. The apex had several layers of small cells, under which there was a group of small but irregularly arranged cells which passed into the rib meristem. The shoot apices of plants transferred from a long to a short-day regime at different time intervals after fulfilling the requirements of minimal photoperiodic induction thus, on the short day, display morphological and anatomical characteristics of various degrees of transition from generative to vegetative stage.     

INFLORESCENCE DEVELOPMENT IN BRASSICA CAMPESTRIS L.

Vegetative seedlings of the Ceres strain Brassica campestris L., a quantitative, long-day plant, were induced to flower by exposure to a 16-hr, long-day cycle. Cytohistological and cytohistochemical changes associated with inflorescence development were examined. Developing shoot apices were classified in vegetative, transitional, and reproductive stages. The vegetative apex possessed a biseriate tunica, central zone, peripheral zone and pith-rib meristem. The transitional stage at 48 hr was marked by an increase in size and by a stratification of the upper cell layers of the shoot apex with a concurrent decrease of apical cytohistochemical zonation. The reproductive stage was initiated at 58 hr by periclinal cell divisions in the 3rd and 4th cell layers of the flank region. Cytohistochemical zonation in the vegetative apical meristem was restored in the floral apex. An “intermediate developmental” phase was not observed between the vegetative and reproductive stage.     

AXILLARY AND DICHOTOMOUS BRANCHING IN THE PALM CHAMAEDOREA

In both Chamaedorea seifrizii Burret and C. cataractarum Martius each adult foliage leaf subtends one axillary bud. The proximal buds in C. seifrizii are always vegetative, producing branches (= new shoots or suckers); and the distal buds on a shoot are always reproductive, producing inflorescences. The prophyll and first few scale leaves of a vegetative branch lack buds. Transitional leaves subtend vegetative buds and adult leaves subtend reproductive buds. Both types of buds are first initiated in the axil of the second or third leaf primordia from the apex, P2 or P3. Later development of both types of bud tends to be more on the adaxial surface of the subtending leaf base than on the shoot axis. Axillary buds of C. cataractarum are similarly initiated in the axil of P2 or P3 and also have an insertion that is more foliar than cauline. However, all buds develop as inflorescences. Vegetative branches arise irregularly by a division of the apex within an enclosing leaf (= P1). A typical inflorescence bud is initiated in the axil of the enclosing leaf when it is in the position of P2 and when each new branch has initiated its own P1. No scale leaves are produced by either branch and the morphological relationship among branches and the enclosing leaf varies. Often the branches are unequal and the enclosing leaf is fasciated. The vegetative branching in C. cataractarum is considered to be developmentally a true dichotomy and is compared with other examples of dichotomous (= terminal) branching in the Angiospermae.     

Growth and Histological Changes During Transition to Dormancy in the Regeneration Buds of Hordeum bulbosum L.

Growth and histological changes in a regeneration bud of Hordeumbulbosum during transition to dormancy were studied. Activeformation of new-leaf primordia on the elongating apex accompaniedby arrest of cell division in the lower leaf primordia characterizedthe first period. When activity subsequently in the distal partof the bud decreased, exillary buds and root primordia werestill actively being produced in its basal part.     

The cell cycle in the shoot apex ofSilene during the first day of floral induction

Summary The length of the cell cycle was measured in the shoot apical meristem ofSilene coeli-rosa during the first day of an inductive photoperiod. The length of the cell cycle in the shoot apex of vegetative controls (those in short days) was about 18–20 hours. Exposure of plants to the long day resulted in an immediate shortening of the cell cycle to about 13 hours, roughly two thirds of that in short days. Measurements of the component phases of the cell cycle revealed that the shortened cycle in long days was the result of a decrease in the length of G 1 and perhaps S, whilst G 2 and M remained constant.     

Rates of Growth and Cell Division in the Shoot Apex of Silene During the Transition to Flowering

The growth rates of the shoot apex during and after floral inductionwere measured in Silene, a long-day plant. Plants were inducedto flower with 4 or more long days (LD) but not with 3 longdays or with short days (SD). The rate of increase of cell numberin the apical dome, above the youngest leaf pair, was exponentialand in plants given 3 LD remained the same as in plants in SD.In plants induced to flower with 7 LD, until the end of theinductive period the rate of increase of cell number in theapical dome remained the same as in plants in SD. Only whenthe apex began to enlarge as the first stage in the formationof the flower did the growth rate of the apical dome increase.The rates of increase of cell numbers in the apex correspondedto mean cell generation times of 20 to 33 h for plants in SD,for plants given 3 LD, and during the 7 days of induction forplants given 7 LD, and 6 to 8 h for induced plants when flowerformation was beginning. The distribution of cell division in the apex was examined bytreating plants with colchicine and noting in sections the positionsof the resulting metaphases. In vegetative apices and also inapices undergoing transition to flowering the whole of the apicaldome appeared to consist of cells dividing at a similar rate. The rate of leaf initiation during induction was the same asin vegetative, non-induced plants.