The effect of lateral illumination on growth oscillations of pine seedling hypocotyls (Pinus silvestris L.)

Phototropic sensitivity of forest wood seedlings to lateral illumination was proposed as an early assay for distinquishing various ecotypes of woody plants of the same species. Statistical analysis showed that results were significantly influenced by heterogenity of experimental material caused by an interference of phototropic movements and natural oscillations of hypocotyl. Both movements of pine seedlings (Pinus silvestris L.) were registered by phase photography and their mechanism was analyzed. The apical part of growing hypocotyl illuminated from above oscillates in a space spiral with frequency 3.3 h at mean growth rate 0.66 mm h^?1. The mean size of spiral amplitudes is 2.9 mm. The oscillation rhythm is disturbed after the lateral illumination and a phase shift was observed as a result. A new oscillation rhythm with frequency 3.9 h and mean growth rate 0.69 mm h^?1 was stabilized after a period of time equal to one nutation turn. Oscillation amplitudes were increased to 4.3 mm. In addition to the radiation intensity the appearance of the phototropic response to light pulse is first of all effected by the actual position of the apical part of hypocotyl in the course of endogenous circumnutations. A uniform plant material for the early assay may be obtained by selection of seedlings which are at the beginning of lateral illumination in the same phase of nutation rhythm. Under such conditions the deviations of longitudinal axis of oscillating spirals characterize the actual phototropic curvature.     

Growth and nutation parameters of primary root of pedunculated oak (Quercus robur L.)

The elongation of primary roots of pedunculated oak (Quercus robur L.), which is associated with nutation oscillations, was registered by a time lapse cinematography. The following characteristics were expressed graphically: relative growth rate [mm h-¹], frequency [rev. h-¹] and amplitudes [mm] of nutation oscillations, their direction and location of nutation curvatures. The growth-oscillation mechanism is interpreted as a part of the geocontrol system of the root and was evaluated on the basis of its physiological efficiency. Comparison of nutation periods showed that the control system of pedunculated oak is three times more and two times less efficient than that of pea and maize, respectively. Comparison of the oscillation amplitudes of growing roots gives similar results.     

Circumnutation oscillations of the hypocotyl and hypocotyl hook formation: A comparison of Norway spruce (Picea abies (L.)Karst.) and Scots pine (Pinus silvestris L.) seedlings

Circumnutation oscillations of hypocotyls were studied in Norway spruce and Scots pine using time-lapse cinematography. It was found that the circumnutations were specific in species. The analysis of hypocotyl ontogeny (100 h) revealed a different duration of the phases I, II and III of nutation oscillations in the first and second taxon. From the quantitative point of view it can be stated that during the phase I of nutation oscillations the Norway spruce hypocotyls exhibited higher frequency activity (corrections in direction of the hypocotyl by an elongation-controlling system corrected by a feedback mechanism), while in the phase III of nutation oscillations higher growth rate and larger amplitudes were observed in the Scots pine hypocotyl when compared with those of Norway spruce. The hypocotyl hook appeared only in Norway spruce.     

Interactions of photo- and geotropism with periodical oscillations of growing pea root (Pisum sativum L.)

Oscillation movements of primary roots of pea seedlings were investigated after low- and high-energy irradiation (10^?2 and 10⁶ W m^?2) which was applied continuously and in pulses at intervals of 3 and 30 min. Oscillation amplitudes of control roots grown in darkness were lower (1 mm) than those of irradiated ones recorded in our previous experiments. The elongation rate of roots grown under scattered and low-energy irradiation as well as their mean oscillation amplitude (2.6 mm) and frequency (8 h) correspond to the standard values recorded in previous experiments. The same effects on root growth and oscillations had scattered electronic flash irradiation. Side-irradiation did not influence the oscillation movements of growing roots. Their frequency and oscillation amplitudes were not changed. Their longitudal axis was, however, deflected 2.75° from the vertical one due to the negative phototropic response of the root. The growth oscillations may be, according to present results, interpreted as consequences of tropic growth controlling system. External factors, like irradiation, may influence oscillation parameters,i.e. their amplitudes and after application of side-irradiation also the direction of longitudinal axis of oscillation spirals. The resultant trajectory is composed from movements which resulted from geotropic and phototropic responses of the roots.     

Nutation rhythm of growing pine hypocotyl (Pinus silvestris L.) interferred with a phototropic stimulus

The interaction of nutation oscillation and phototropic curvature mechanisms of growing pine hypocotyl(Pinus silvestris L.) was investigated. Nutation parameters of growing hypocotyl are not affected by continuous lateral illumination. Frequency and nutation amplitudes undergo changes which are dependent on plant development and are nearly the same when plants are illuminated from one side or from above. Lateral illumination, however, induces a phase shift. This deviation from a normal nutation rhythm usually disappears after a period of time equal to three nutation revolutions,i.e. after 12 h. The positive phototropio response of nutating hypocotyl is dependent on the phase in which the hypocotyl is exposed to the lateral illumination:(a) nutation perihelium,(b) ahelium and(c) nutation interphase. These response-variants are compared with the expected gradients of plant hormones at the opposite sites of the hypocotyle. Analysis of location of the individual movement reactions(i.e. oscillations as a component of the geocontrol system and phototropic curvature) together with the determination of the zone of maximum elongation indicates the existence of different receptory centres of these two movement systems. In the case of the phototropic response reaction it is possible to consider the presence of different photo- and biochemical mechanisms at least for a part of this system.     

An auxin-resistant mutant ofNicotiana plumbaginifolia Viv. is impaired in 1-naphthaleneacetic acid-induced hyperpolarization of hypocotyl cell membranes in intact seedlings

The electrical response to the synthetic auxin 1-naphthaleneacetic acid (1-NAA) ofNicotiana plumbaginifolia wild type and the monogenic, dominant auxinresistant mutant R25 was studied. Membrane potentials were continuously recorded in hypocotyl cells of light-grown, intact seedlings, and the time course of the response to 1-NAA addition was followed. Wild-type cells responded to ? 10^?5 M 1-NAA with a delayed, transient hyperpolarization. The R25 cells hyperpolarized significantly only in response to 1-NAA at 10^?3 M, and with maximal amplitudes lower than those recorded with the wild type. In contrast, the two genotypes reacted similarly in terms of kinetics and amplitude to 10^?5 M fusicoccin, which rapidly and strongly hyperpolarized the cells, and to 10^?3 M benzoic acid, which induced rapid and weak hyperpolarization. The resting membrane potentials of the wild type and R25 were also not significantly different. Unlike wild-type hypocotyls, those of R25 ceased elongating before the time chosen for the electrophysiological measurements, but control experiments performed at a time when the elongation of both genotypes had terminated indicated that the difference in electrical response to auxin is independent of hypocotyl growth. The inefficiency of 1-NAA in inducing hyperpolarization of R25 hypocotyl cells suggests a defect at an early step in auxin action.     

Diurnal oscillatory movements of growing leaves of tobacco

The analysis of diurnal oscillatory movements of tobacco leaves was used in the diagnosis of viral infection of plants. The oscillatory helices circumscribed by a growing leaf of a healthy plant were regular, but some deviations, particularly in the transition points, were recorded. In order to make clear the cause of these irregularities of trajectory, the course of elongation of leaf petiole and blade in relation to localization and shift of zones of elongation during ontogenesis was analysed. The present analysis is similar to that described by the author's earlier experiments with pea roots. Oscillatory curves circumscribed by petiole, projected on a horizontal plane, were compared with curves circumscribed by the blade tip. The analysis of the leaves of different ages enabled us to study this process in dependence on growth rate. It was confirmed that oscillations are a result of elongation; the extent of oscillations is quantitatively dependent on the growth rate. An analysis of the zones of growth showed that in petiole the active meristems are localized near to its base while in the leaf lamina they move gradually during the ontogenesis from the apical to the basal part of the leaf blade. Active meristems of young rapidly growing leaves are localized approximately in the middle of the blade while those of old leaves were found in close proximity to the base of the lamina. The growth rate of petiole can be expressed in hundreds of mm per hour (4.8×10^?2 mm h^?1); half of this value was recorded for its apical part. The growth rate of leaf blade was found approximately ten times higher (3.2×10^?1 mm h^?1). The oscillatory movements of growing leaf consists of two integrate components: of oscillations originating in the base of the petiole and of oscillations of leaf blade the centrum of which is localized in the basal third of the blade. The arrangement of the experiments did not enable us to determine to what extent the phototropic response of leaf blade participates in leaf movements. The movements of leaves of an intact plant are evidently affected by rhythmic stem oscillations. Stem is an integral part of a system which participates in the transfer of information in plants.     

Diurnal and ultradiurnal oscillations of growing organs within the framework of the information system of the plant

The analysis of growth and movements of seedling organs of kidney bean (Phaseolus vulgaris L.) provides a pattern of periodic phases of activity and relaxation. The existence of a central organ which would control the phase relationships, is not anticipated in the integrity of the plant. The cyclic activity of individual organs shows itself by growth associated with oscillation movements. One and the same organ may simultaneously accomplish oscillatory movements with a diurnal and ultradiurnal frequency. These rhythms originate during the organ development; the first pair of kidney bean leaves at first executes oscillation movements with a diurnal frequency and only after it is fully developed it exhibits a diurnal cycle with the photophil phase upwards and the scotophil downwards, the oscillations with an ultradiurnal oycle being maintained. The movements of the two leaves are synchronous, but there occur short sections with a desynchronous cycle. Simultaneously with these oscillations, in which the leaf petiole takes part, the adult leaf performs oscillatory movements perpendicular to the longitudinal leaf axis, the so-called side swings, controlled by periodical changes of the joint attaching the leaf blade. Their frequency is practically identical with that of the ultradiurnal cycle. Thus the periodic growth activity of the kidney bean results in growth oscillations passing in the diurnal cycle with a frequency of 0.043 rev.h^-1, their ascending and descending phases consisting of periodical ultradiurnal oscillations in cycles of 0.73–0.59 rev.h^-1. The epicotyl growth shows a similar pattern: into the basic diurnal nutation cycle with a frequency of 0.042 rev.h^-1 ultradiurnal oscillation cycles are incorporated having a similar frequency to that revealed in leaves (0.69–0.64 rev.h^-1). The diurnal oscillatory cycles belong to a system established on the basis of periodicity of day and night and other geophysical cycles. The ultradiurnal rhythmic oscillations are presumed to be an expression of the geocontrol system of root and shoot growth direction and orientation of the organ in space. The shape of their trajectories in bean leaves is contradictory to this; they are not spatial helices, as the kybernetic model would presuppose, but have a vertical, upwards and downwards course in one plane. Since these oscillatory movements with an ultradiurnal cycle cease after petiole excision from the stem and after shoot apex amputation, one may presume that they are coupled with the low-frequency oscillatory system of the epicotyl.     

Signal transmission during gravitropic curvature of primary roots of Zea mays

Primary roots of Zea mays cv. Ageotropic are nonresponsive to gravity and elongate approximately 0.80 mm h^?1. Applying mucilage-like material (K-Y Jelly) to the terminal 1.5 cm of these roots induces graviresponsiveness and slow elongation 28% (i.e. from 0.80 to 0.58mm h^?1). Applying mucilage-like material to one side of the terminal 1.5 cm of the root induces curvature toward the mucilage, irrespective of the root's orientation to gravity. Applying a 2-mm-wideband of mucilage-like material to a root's circumference 8 to 10 mm behind the root cap neither induces gravicurvature nor affects elongation significantly. Similarly, applying mucilage-like material to only the root cap does not significantly affect elongation or graviresponsiveness. Gravicurvature of mutant roots occurs only when mucilage-like material is applied to the root/root-cap junction. Reversing the caps of wild-type and mutant roots produces gravitropic responses characteristic of the root cap rather than the host root. These results are consistent with the suggestion that gravitropic effectors are growth inhibitors that move apoplastically through mucilage between the root cap and root.     

Effect of root tip amputation on spiral oscillations of the growing hypocotyl with radicle of the pea (Pisum sativum L.)

In order to interpret the mechanism of elongation growth of the hypocotyl with radicle in the first stages of germination of the pea seed (Pisum sativum L.) a cybernetic model utilizing feed-back as a mechanism of correcting the wrong direction of growth was proposed (Spurný 1966, 1967). In the present study, the effect of amputation of the root tip as the control centre on the trajectory of the growing radicle was investigated. The results have shown that the hypocotyl grows, elongates—the rate of growth being slightly lower than that of the standard, but that no spiral oscillations at all are executed by the organ after amputation of the root tip. This finding appears to confirm the applicability of the proposed cybernetic model, for amputation of the root tip means that not only the control block is eliminated, but also that the channel of feed-back impulses to zone of elongation is interrupted.     

GROWTH PATTERNS IN NUTATING AND NONNUTATING SUNFLOWER (HELIANTHUS ANNUUS) HYPOCOTYLS

Dark-grown sunflower (Helianthus annuus L.) seedling hypocotyls (15–30 mm) were marked with two rows of lanolin-coated resin beads, and the events of the following 24 hr, in physiological darkness, were recorded on time-lapse video. Nutational movement of the hypocotyl, followed for 20–24 hr for each of 21 seedlings, was found to have a mean period of 153 ± 26 min (ca. 24 C). Displacement of each bead, with time, was measured with a microcomputer-controlled video analyzer, and relative elemental elongation rate and relative growth rate analyses were carried out to determine the spatial and temporal distribution of growth. Relative elemental elongation rates were plotted against distance and time to produce “growth landscapes.” A strongly nutating seedling showed periodic fluctuations in local growth rates that alternated between values of 0.0 hr^−-1 and >0.12 hr^−-1 near the hypocotyl hook. Nearer the base, maximum growth rates were lower but local periodic changes still were evident. Seedlings, in which nutation appeared during the time period analyzed, showed non-synchronous pulses of growth along the axis. With nutational development, these local growth fluctuations became synchronized along each side and phased (usually 180° out of phase) with the coordinated growth fluctuations along the opposite side. In some seedlings the changes from low to high local growth rates occur nearly simultaneously over two-thirds of the active region. In others, basipetally traveling waves of growth are suggested by the growth landscapes.     

Influence of cobalt on soybean hypocotyl growth and its ethylene evolution

Development of dark-grown “Clark” soybean (Glycine max [L.] Merr.) seedlings is abnormal at 25 C but normal at 20 and 30 C. At 25 C, hypocotyls swell and fail to elongate normally; lateral root formation and seedling ethylene evolution are enhanced.

Co²⁺ promoted hypocotyl elongation of etiolated “Clark” soybean seedlings by 28% when grown at 25 C. The same growth-promoting concentration reduced hypocotyl thickness and primary root elongation by 28 and 43%, respectively. Co²⁺ inhibited ethylene production both of intact seedlings and of apical 1-centimeter hypocotyl segments with attached epicotyls and cotyledons by 65 and 60%, respectively. These results suggest that Co²⁺ exerts its effects on the hypocotyl growth by inhibiting ethylene production, and also confirm our previous conclusion that abnormal ethylene production at 25 C is responsible for the inhibition of hypocotyl elongation and for its swelling.

     

The changes in the growth pattern of organs ofChenopodium rubrum photoperiodically induced to flowering

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.     

Analysis of Cell Division and Elongation Underlying the Developmental Acceleration of Root Growth in Arabidopsis thaliana

To investigate the relation between cell division and expansion in the regulation of organ growth rate, we used Arabidopsis thaliana primary roots grown vertically at 20°C with an elongation rate that increased steadily during the first 14 d after germination. We measured spatial profiles of longitudinal velocity and cell length and calculated parameters of cell expansion and division, including rates of local cell production (cells mm⁻¹ h⁻¹) and cell division (cells cell⁻¹ h⁻¹). Data were obtained for the root cortex and also for the two types of epidermal cell, trichoblasts and atrichoblasts. Accelerating root elongation was caused by an increasingly longer growth zone, while maximal strain rates remained unchanged. The enlargement of the growth zone and, hence, the accelerating root elongation rate, were accompanied by a nearly proportionally increased cell production. This increased production was caused by increasingly numerous dividing cells, whereas their rates of division remained approximately constant. Additionally, the spatial profile of cell division rate was essentially constant. The meristem was longer than generally assumed, extending well into the region where cells elongated rapidly. In the two epidermal cell types, meristem length and cell division rate were both very similar to that of cortical cells, and differences in cell length between the two epidermal cell types originated at the apex of the meristem. These results highlight the importance of controlling the number of dividing cells, both to generate tissues with different cell lengths and to regulate the rate of organ enlargement.     

Stimulation of root hair elongation in Arabidopsis thaliana by low phosphorus availability

Low phosphorus availability stimulates root hair elongation in many plants, which may have adaptive significance in soil phosphorus acquisition. We investigated the effect of low phosphorus on the elongation of Arabidopsis thaliana root hairs. Arabidopsis thaliana plants were grown in plant culture containing high (1000 mmol m^?3) or low (1 mmol m^?3) phosphorus concentrations, and root hair elongation was analysed by image analysis. After 15d of growth, low-phosphorus plants developed root hairs averaging 0.9 mm in length while high-phosphorus plants of the same age developed root hairs averaging 0.3 mm in length. Increased root hair length in low-phosphorus plants was a result of both increased growth duration and increased growth rate. Root hair length decreased logarithmically in response to increasing phosphorus concentration. Local changes in phosphorus availability influenced root hair growth regardless of the phosphorus status of the plant. Low phosphorus stimulated root hair elongation in the hairless axr2 mutant, exogenously applied IAA stimulated root hair elongation in wild-type high-phosphorus plants and the auxin antagonist CM PA inhibited root hair elongation in low-phosphorus plants. These results indicate that auxin may be involved in the low-phosphorus response in root hairs.     

A novel optical interference technique to measure minute root elongations of Japanese red pine (Pinus densiflora Seibold & Zucc.) seedlings infected with ectomycorrhizal fungi

This study presents a new technique to measure root elongation of Japanese red pine (Pinus densiflora Seibold & Zucc.) seedlings with very high sensitivity in the order of sub-nanometer by using a novel optical interference method called statistical interferometry. The principle of the statistical interferometry is based on the statistics of a speckle field, which is generated when a rough surface is illuminated by a laser light. The technique facilitates to obtain minute root elongation measurements in the order of sub-seconds. The root elongation behavior of Pinus densiflora seedlings infected with ectomycorrhizal fungi, Pisolithus sp. (Ps) and Cenococcum geophilum Fr. (Cg), was investigated in comparison with that of an uninfected control. In the experiments, two points on a root with the separation of 3 mm were illuminated by laser beams and the elongation was measured continuously by analyzing speckle patterns successively taken by a CCD camera. The root elongation rate (RER), measured as the length of root elongation per second per millimeter (mean ± S.D.) for Ps-infected, Cg-infected and uninfected seedlings were 10.85 ± 2.41, 5.54 ± 1.43, and 2.41 ± 1.01 nm s^?1 mm^?1, respectively. We found that the RERs of seedlings infected with ectomycorrhizal fungi were significantly higher than that of the uninfected seedlings, and the seedlings infected with Ps fungi showed the highest RER. We conducted another experiment to observe two-dimensional root growth, in which the growth measurements were obtained for 4 months. From this experiment, we observed that root growth of ectomycorrhizal fungi infected seedlings were higher than that of the uninfected seedlings. The evaluation of results from these two techniques proves that the proposed statistical interferometry is much faster and very sensitive technique, where the time required for growth monitoring is 10⁷ times less than the other. We can conclude that, at the scale of either very short time or relatively long time, the symbiotic relationship between root and ectomycorrhiza has a positive effect steadily on the root elongation.     

I. Cytochrome oscillations in continuous culture: Action of chloramphenicol on Saccharomyces carlsbergensis and Candida utilis

Oscillations in the A, B and C group cytochromes of Saccharomyces carsbergensis and Candida utilis have been observed in continuous culture following the addition and removal of chloramphenicol. With S. carlsbergensis reproduction of these oscillations proved difficult due to the fermentative nature of the yeast. With C. utilis at D = 0.1 h⁻¹ the oscillations in teh A group cytochromes were statistically significant and 3–6 times the standard deviation observed in steady sate. The oscillations provide evidence of a feedback loop controlling cytochrome synthesis and have a period of the same order of that predicted by Goodwin for the protein (enzyme) component of his model (viz. 6–8 h). With C. utilis the period of these oscillations was a function of the growth rate varying from approx. 10 h at a growth rate of 0.1 h⁻¹ (mean generation time 6.9 h) to 3 h at a growth rate of 0.3 h⁻¹ (mean generation time 2.3 h).     

Spiral feedback oscillations of growing hypocotyl with radicle inPisum sativum L.

Cinematographic records of longitudinal growth showed that hypocotyl with radicle inPisum sativum L. undergoes spiral oscillations during growth. This phenomenon can be characterized by the following time-space limits:

1. (1)
   Curvature of the hypocotyl with radicle takes place always in the zone of most rapid elongation (Fig. 2, 8).     
   
   

Gibberellin-Dependent Root Elongation in Lactuca sativa: Recovery from Growth Retardant-Suppressed Elongation with Thickening by Low concentration of GA3

Higher concentrations of growth retardants, ancymidol and AMO-1718,were required to suppress root growth than hypocotyl growthin lettuce seedlings. Gibberellic acid (GA₃) counteracted theeffect of these growth retardants, but complete recovery ofroot growth was obtained only in a narrow range of growth retardantconcentrations. A much lower concentration of GA₃ (1 nM) wasneeded for recovery of root growth from ancymidol suppressionthan that for hypocotyl growth (100 (µM). GA₃ synergisticallypromoted root growth at moderate concentrations (10–100nM) with either ancymidol or AMO-1618. Ancymidol not only suppressed root elongation but also causedthickening of the elongation zone of the root, actions whichGA₃ completely canceled. Microscopic observation showed theseeffects were mainly due to the lateral expansion and shorteningof epidermal and cortical cells. Growth kinetics of roots recorded by a computer-regulated rhizometerindicated that the lag times of both growth suppression by ancymidoland growth recovery by GA₃ were about 4 h. -Naphthaleneacetic acid (NAA) did not counteract ancymidol suppressionof root and hypocotyl growth. These results support the concept that gibberellins play anindispensable role in root elongation at an extremely lowerconcentration than in hypocotyl elongation. (Received January 7, 1987; Accepted May 14, 1987)     

Diurnal changes in shoot water dynamics are synchronized with hypocotyl elongation in Arabidopsis thaliana

We recently demonstrated the circadian clock modulated water dynamics in the roots of a small model plant, Arabidopsis thaliana, by the Nuclear Magnetic Resonance (NMR) microimaging technique. Our developed technique was able to visualize the water distribution that depended on differences in the ¹H signal among region in the shoot, such as the shoot apex, the hypocotyl and the root shoot junction. Water content in the shoot increased during periods of light in comparison with dark periods, and continued through the early stage of seedling growth until the dark period. When the water content changed, elongation and/or movement occurred in the hypocotyl, and these events were synchronized. The water dynamics of the shoot also displayed an opposite phase with the root water dynamics.