Changes in membrane potential inChenopodium rubrum during the course of photoperiodic flower induction

Electrophysiological processes were investigated in the reception organ of photoperiodism, cotyledons and first leaves, in a model short-day plantChenopodium rubrum L. (selection 374) within the dark inductive cycle for flowering. Membrane potential (E_m) was measured in cotyledon and first leaf mesophyll of intact plants. The E_m time-course was fairly similar during inductive dark or postinductive light period or in non-inductive continuous light and had a character of irregular oscillations. The most distinct oscillations were found during the postinductive light period. Changes in light régime at the beginning (light off) and the end of inductive dark period (light on) triggered marked transient E_m changes having a character of damped oscillations. Cortical root cells in intact plants did not react to switching light and darkness. Changes in E_m in reception organs during the inductive cycle could not be correlated with the formation and transport of floral stimulus or with reaching the induced state. Thus, the electrophysiological nature of floral stimulus has not been confirmed.     

The comparison between membrane and transorgan electric potentials inChenopodium rubrum: The methods

A new method of simultaneous measurement of membrane (E_m) and transorgan (E_tr)electric potential in intactChenopodium rubrum plants and changes in E_m and E_tr under various experimental conditions are described. E_m was measured in mesophyll leaf cells, and E_tr in the same plant as a potential difference between a first pair leaf, tip and roots. The two potentials differed distinctly, E_m averaging-156mV and E_tr -5mV. But the,-changes in E_m and E_tr had approximately the same magnitude and time-course after changing light and darkness or KCl concentration in solution flowing around the leaf or during the photoperiodic induetive cycle. In relation to the same electrode connection they had an opposite polarity. The nature of E_tr and its relation to E_m are discussed. The measurement of E_tr is much more stable and simpler than that of E_m. The similarity of E_m and E_tr time-courses justifies the use of E_tr measurement in electrophysiological studies as a replacement for E_m measurements especially in intact plants.     

Studies on the Transmission of Floral Effects of Photoperiod and Gibberellin from One Branch to the Other inImpatiens balsamina

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.     

Hydraulic architecture of sugarcane in relation to patterns of water use during plant development*

Hydraulic conductance was measured on leaf and stem segments excised from sugarcane plants at different stages of development. Maximum transpiration rates and leaf water potential (Ψ_L) associated with maximum transpiration were also measured in intact plants as a function of plant size. Leaf specific hydraulic conductivity (L_sc) and transpiration on a unit leaf area basis (E) were maximal in plants with approximately 0.2 m² leaf area and decreased with increasing plant size. These changes in Fand L_sc were nearly parallel, which prevented φ_L in larger plants from decreasing to levels associated with substantial loss in xylem conductivity caused by embolism formation. Coordination of changes in E and leaf hydraulic properties was not mediated by declining leaf water status, since φ_L increased with plant size. Hydraulic constrictions were present at nodes and in the node-leaf sheath-leaf blade pathway. This pattern of constrictions is in accord with the idea of plant segmentation into regions differing in water transport efficiency and would tend to confine embolisms to the relatively expendable leaves at terminal positions in the pathway, thereby preserving water transport through the stem.     

Effect of varying lengths of inductive and supplementary non-inductive photoperiods on vegetative growth and flowering of Impatiens balsamina

The photoperiodic requirement for flowering in Impatiens balsaminachanges with the length of the photoperiod. Floral buds wereinitiated with two 8 hr but with four 15 hr photoperiods andflowers opened with four 8 hr but twenty-eight 15 hr photoperiods.A part of the photoperiodic requirement for floral inductionin this plant can be substituted by LDs containing 4 or morehours of darkness (10). It indicates the identical nature ofthe floral stimulus produced during the dark period, whetherit forms a part of the inductive or non-inductive cycles. Theeffect of these supplementary non-inductive photoperiodic cyclesin causing floral bud initiation also depends on the lengthof the first inductive obligatory cycle. More floral buds andflowers were produced on plants exposed to 15 hr than 8 hr photoperiods,probably due to the higher number of leaves that were producedunder the former condition of weaker induction. The shorterthe dark period in the photoperiodic cycle, the weaker the induction,the slower the rate of extension growth but the more differentiationof leaves. ¹ Present address: Department of Biology, Guru Nanak Dev University,Amritsar-143005, India. (Received November 9, 1977; )     

Puzzle box,a tobacco line with flowers that mix floral and inflorescence characteristics

We characterized the development of a tobacco morphological mutant, puzzle box, previously obtained by selection for resistance to an inhibitor of an enzyme in the polyamine biosynthetic pathway. Puzzle box plants are shorter than wild type with smaller leaves and an irregular leaf plastochron. The diameter and shape of the puzzle box shoot apex are similar to wild type. In puzzle box plants, the terminal flower develops but not the cymose inflorescence. This flower has a variable phenotype with more than five sepals, five petals, and five stamens. The organs in the fourth whorl are produced by carpellike primordia and contain tissue biochemically similar to wild type transmitting tissue. These organs form a cylinder within which additional floral organs are produced. Some cultures of puzzle box buds and excised cylinders produce additional floral organs. The inflorescence and floral programs may be expressed together in puzzle box.     

Rapid interorgan communications in higher plants with special reference to flowering

Several examples of transmission of rapid signals within plants are described. Most of these signals may be inhibited by pretreating plants with LiCl, with inhibitors of ionic permeability, or with substances interfering with Ca²⁺. Accordingly, they are dependent on ionic changes. LiCl also prevents or delays the onset of flowering in long-day plants. In addition isolated organs or tissues most often do not react to an external stimulus as do similar organs or tissues within an intact plant. All these facts lead to the conclusion that plants are highly integrated organisms. Any change in the ionic equilibrium occurring in one part of these organisms has rapid consequences in other parts. This hypothesis may be applied to the floral process: after perception of the appropriate environmental conditions, the leaves modify the orientation of meristem differentiation by promoting the redistribution of ions (especially Ca²⁺, which has a “second messenger” function).     

Non-reversion of Impatiens in the absence of meristem commitment

Purple-flowered plants of Impatiens balsamina maintained floral development on transfer from inductive short days (SD) to long days (LD), a treatment in which red-flowered plants of Impatiens are known to revert to leaf production. An investigation into the non-reverting nature of purple-flowered plants was carried out to establish whether these plants achieved meristem commitment or whether their non-reverting state was controlled by the leaves. When the leaves that had unfolded during the inductive SD treatment were removed at the time of transfer to LD, the purple-flowered plant did revert. This result suggests that, as in red-flowered Impatiens, meristem commitment is absent, but that purple-flowered plants maintain flowering in LD conditions because of a more permanent supply of signal from their leaves than occurs in red-flowered plants. A working hypothesis is proposed to explain how a signal from the leaves can retain a controlling role during flower development.Key words: Floral commitment, Impatiens, floral reversion, floricaula.      

Promotory effect of GA13 on flowering ofAmaranthus — a short day plant

Abstact 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.     

Auxin-induced leaf blade expansion in Arabidopsis requires both wounding and detachment

Elevation of leaf auxin (indole-3-acetic acid; IAA) levels in intact plants has been consistently found to inhibit leaf expansion whereas excised leaf strips grow faster when treated with IAA. Here we test two hypothetical explanations for this difference in growth sensitivity to IAA by expanding leaf tissues in vivo versus in vitro. We asked if, in Arabidopsis, IAA-induced growth of excised leaf strips results from the wounding required to excise tissue and/or results from detachment from the plant and thus loss of some shoot or root derived growth controlling factors. We tested the effect of a range of exogenous IAA concentrations on the growth of intact attached, wounded attached, detached intact, detached wounded as well as excised leaf strips. After 24 h, the growth of intact attached, wounded attached, and detached intact leaves was inhibited by IAA concentrations as little as 1 µM in some experiments. Growth of detached wounded leaves and leaf strips was induced by IAA concentrations as low as 10 µM. Stress, in the form of high light, increased the growth response to IAA by leaf strips and reduced growth inhibition response by intact detached leaves. Endogenous free IAA content of intact attached leaves and excised leaf strips was found not to change over the course of 24 h. Together these results indicate growth induction of Arabidopsis leaf blade tissue by IAA requires both substantial wounding as well as detachment from the plant and suggests in vivo that IAA induces parallel pathways leading to growth inhibition.     

Determination for inflorescence development is a stable state,separable from determination for flower development in Pisum sativum L. buds

Terminal meristems of Pisum sativum (garden pea) transit from vegetative to inflorescence development, and begin producing floral axillary meristems. Determination for inflorescence development was assessed by culturing excised buds and meristems. The first node of floral initiation (NFI) for bud expiants developing in culture and for adventitious shoots forming on cultured meristems was compared with the NFI of intact control buds. When terminal buds having eight leaf primordia were excised from plants of different ages (i.e., number of unfolded leaves) and cultured on 6-benzylaminopurine and kinetin-supplemented medium, the NFI was a function of the age of the source plant. By age 3, all terminal buds were determined for inflorescence development. Determination occurred at least eight nodes before the first axillary flower was initiated. Thus, the axillary meristems contributing to the inflorescence had not formed at the time the bud was explanted. Similar results were obtained for cultured axillary buds. In addition, meristems excised without leaf primordia from axillary buds three nodes above the cotyledons of age-3 plants gave rise to adventitious buds with an NFI of 8.3 ±0.3 nodes. In contrast seed-derived plants had an NFI of 16.5 ±0.2. Thus cells within the meristem were determined for inflorescence development. These findings indicate that determination for inflorescence development in P. sativum is a stable developmental state, separable from determination for flower development, and occurring prior to initiation of the inflorescence at the level of meristems.     

Gas Exchange and Phytoluminography of Single Red Kidney Bean Leaves during Periods of Induced Stomatal Oscillations: A Demonstration of an Integrated, Spatially Resolving Physiometric Technique

This report examines the capabilities of a new approach to the study of gas exchange and electron transport properties of single, intact leaves. The method combines conventional aspects of analysis with an image intensification system that records the spatial distribution of delayed light emission (DLE) over single leaf surfaces. The combined system was used to investigate physiological perturbations induced by exposure of single leaves of Phaseolus vulgaris cv `California Light Red' to a combination of SO₂ (0.5 microliters per liter) and ozone (0.1 microliters per liter). Exposure of one-half of a leaf to this combination induced DLE and stomatal oscillations, but only in the half of the leaf exposed to the combined gases. Examination of phytoluminographs taken during these oscillations revealed distinct leaf patches where the greatest changes in DLE intensity occurred. This phenomenon is interpreted to be evidence that control of stomatal activity of intact plant leaves occurs within discrete leaf areas defined within the vascular network.     

Characterization of leaf senescence and pod development in soybean explants

Excised soybean (Glycine max [L.] Merrill) cv Anoka leaf discs tend to remain green even after the corresponding intact leaves have turned yello on fruiting plants. We have found that explants which include a leaf along with a stem segment (below the node) and one or more pods (maintained on distilled H₂O) show similar but accelerated leaf yellowing and abscission compared with intact plants. In podded explants excised at pre-podfill, the leaves begin to yellow after 16 days, whereas those excised at late podfill begin to yellow after only 6 days. Although stomatal resistances remain low during the first light period after excision, they subsequently increase to levels above those in leaves of intact plants. Explants taken at mid to late podfill with one or more pods per node behave like intact plants in that pod load does not affect the time lag to leaf yellowing. Explant leaf yellowing and abscission are delayed by removal of the pods or seeds or by incubation in complete mineral nutrient solution or in 4.6 micromolar zeatin. Like chorophyll breakdown, protein loss is accelerated in the explants, but minerals or especially zeatin can retard the loss. Pods on explants show rates and patterns of color change (green to yellow to brown) similar to those of pods on intact plants. These changes start earlier in explants on water than in intact plants, but they can be delayed by adding zeatin. Seed dry weight increased in explants, almost as much as in intact plants. Explants appear to be good analogs of the corresponding parts of the intact plant, and they should prove useful for analyzing pod development and mechanisms of foliar senescence. Moreover, our data suggest that the flux of minerals and cytokinin from the roots could influence foliar senescence in soybeans, but increased stomatal resistance does not seem to cause foliar senescence.     

Floral stimulus movement in perilla and flower inhibition caused by noninduced leaves

Photoperiodic control of flowering in the short day plant Perilla involves the transmission of a floral stimulus from induced leaves to the shoot apex. We have studied the basipetal movement of this stimulus and of ¹⁴C-labeled assimilates in plants with an induced leaf (donor) grafted into the uppermost internode of a vegetative plant in which the axillary shoots at various nodes along the stem function as receptors.     

Juvenility and flowering of Brunonia australis (Goodeniaceae) and Calandrinia sp. (Portulacaceae) in relation to vernalization and daylength

Background and Aims

The time at which plants are transferred to floral inductive conditions affects the onset of flowering and plant morphology, due to juvenility. Plants of Brunonia australis and Calandrinia sp. were used to investigate whether Australian native ephemeral species show a distinct juvenile phase that can be extended to increase vegetative growth and flowering.

Methods

The juvenile phase was quantified by transferring seedlings from less inductive (short day and 30/20°C) to inductive (vernalization or long day) conditions at six different plant ages ranging from 4 to 35 d after seed germination. An increase in days to first visible floral bud and leaf number were used to signify the end of juvenility.

Key Results

Brunonia australis was receptive to floral inductive long day conditions about 18–22 d after seed germination, whereas plants aged 4–35 d appeared vernalization sensitive. Overall, transferring plants of B. australis from short to long day conditions reduced the time to anthesis compared with vernalization or constant short day conditions. Calandrinia sp. showed a facultative requirement for vernalization and an insensitive phase was not detected. Floral bud and branch production increased favourably as plant age at time of transfer to inductive conditions increased. Younger plants showed the shortest crop production time.

Conclusions

Both species can perceive the vernalization floral stimulus from a very young age, whereas the photoperiodic stimulus is perceived by B. australis after a period of vegetative growth. However, extending the juvenile phase can promote foliage development and enhance flower production of both species.     

Genetic Regulation of Development in Sorghum bicolor: IV. GA(3) Hastens Floral Differentiation but Not Floral Development under Nonfavorable Photoperiods

Sorghum bicolor (L.) Moench lines with genetic differences in photoperiod requirement were planted in the field near Plainview, Texas (about 34° northern latitude) around June 1 and treated with gibberellic acid (GA₃) solutions applied in the apical leaf whorl. GA₃ hastened the date of floral differentiation (initiation). The greatest responses to GA₃ were by 90M and 100M, the latest of the genotypes, for which floral initiation dates were hastened an average of 19.5 and 21.7 days, respectively, for the 4 years beginning in 1980. There were very small differences in dates of anthesis between control and GA₃-treated plants. Microscopic examination of apical meristems collected between the date of floral initiation of GA₃-treated plants and the later date of initiation of control plants revealed: (a) several morphological characteristics of floral differentiation in the apical meristem of treated plants, (b) consistent occurrence of vegetative morphology in control plants, (c) a few meristems from GA₃-treated plants that appeared to be regressing in floral development and thus possibly exhibiting dedifferentiation. Dedifferentiation of prepanicle primordia into leaves would explain the observed equal or greater number of leaves in GA₃-treated plants rather than the expected smaller number. It is apparent that the presence of a morphological differentiated floral meristem in sorghum does not drive subsequent floral development in the absence of inductive photoperiods. This further suggests that initial floral differentiation and subsequent floral development may be controlled separately in sorghum.     

Effect of ínductive photoperiod and gibberellin treatment on peroxidase enzyme system in relation to floral induction inImpatiens balsamina L

Gibberellins A₃ and A₁₃ cause floral induction inImpatiens balsamina, a qualitative short day plant, under non-inductive 24-h photoperiods (continuous illumination). However, the influence of the two inductive factors,i.e. gibberellins and short days (8-h photoperiods) on the peroxidase enzyme system is different. The total peroxidase activity decreases under both inductive and non-inductive photoperiods, with or without gibberellin treatment. The electrophoretic pattern of isoperoxidases changes only in response to gibberellin treatment. Under 24-h photoperiods, treatment with gibberellins A₃ and A₁₃ causes the appearance in the stem of three additional isoenzymes of peroxidase (Rm 0.50, 0.71 and 0.76). These bands do not appear in the leaves, which are non-essential for gibberellin-caused floral induction in this plant. Under 8-h photoperiods also, gibberellins induce the appearance of new isoenzyme bandsi.e. two in the stem (Rm 0.50 and 0.76) and one in the leaves (Rm 0.05). These may be correlated with the synergistic increase in the number of floral buds in these plants in response to simultaneous exposure to two inductive factors.     

Developmental stability in flowers of Clarkia tembloriensis (Onagraceae)

Developmental instability of floral traits is examined in four populations of Clarkia tembloriensis (Onagraceae) with different natural outcrossing rates. Developmental instability is estimated using fluctuating asymmetry (FA) and within plant variance. The results are coupled with those from a previous study of leaf traits. In the first experiment, flowers were collected from the same growth chamber-grown plants that had been previously used to estimate leaf developmental stability in two C. tembloriensis populations. These populations differed in FA for only one floral trait, long filament length. After adjusting for organ size differences, we found floral FA values were about half those of leaves. These are the first quantitative data indicating that flowers are more developmentally stable than leaves. In a second experiment, greenhouse grown plants from two other C. tembloriensis populations (one highly outcrossing and one predominantly self-pollinating) did not differ significantly in floral FA or in within-plant variance of floral traits, though earlier studies of the same populations revealed significant differences in FA of leaf traits. In both experiments, FA values of different floral traits were uncorrelated. We attribute the lack of significant differences in floral stability between populations to the greater canalization of floral organs and to the magnification of measurement error that occurs when calculating FA. We also found that the shorter styles of selfers are the greatest difference in flower form between predominantly self-pollinating and predominantly outcrossing populations of C. tembloriensis.     

The leaf as the site of gibberellin action in flower formation in Bryophyllum daigremontianum

Summary The long-short-day plant Bryophyllum daigremontianum can be induced to flower by transfer from long to short days (LDSD), or by gibberellin (GA) application under SD. Application of GA to mature leaves of intact or partially defoliated plants induces flowering more effectively than when applications are made to the youngest leaf pair and the shoot tip.Mature leaves on de-budded plants in SD are induced to produce floral stimulus by GA application, as demonstrated by grafting LD receptor scions onto the debudded plants, or by grafting SD leaves treated with GA onto receptor stocks in LD. This shows that GA applied to Bryophyllum in SD exerts its flower-promoting effect in the leaves.The minimal number of SD necessary for flower formation in Bryophyllum is approximately 15, both in case of photoinduction by the shift LDSD, and after GA treatment in SD. It is concluded that the LD part of photinduction establishes a high level of endogenous GAs in the leaves which is a prerequisite for production of floral stimulus under subsequent SD.Work supported by the United States Atomic Energy Commission, Contract No. AT(11-1) 1338.     

EFFECTS OF LEAF EXCISION ON FLOWERING OF XANTHIUM APICAL BUDS IN CULTURE UNDER INDUCTIVE AND NONINDUCTIVE PHOTOPERIODS

Apical buds of Xanthium were grown in aseptic culture under short-day cycles known to induce flowering in the intact plants or under “light-break” conditions known to prevent flowering. The total light provided in each 24-hr cycle was the same under the two photoperiods. Various numbers of leaves were excised from the apical buds. Excision of leaves did not change the response to photoperiod: even with all leaves excised the apical buds cultured under short-day conditions reached the same average floral stage as the control buds, and those under light-break conditions all remained vegetative. Fresh weight was not significantly changed by the excisions, either. However, excision of the young leaves resulted in an increase in the number of new leaves developed by the apical bud during the two-week culture period.