Investigations of the Geotropic Curvature of the Avena Coleoptile

The geotropic reaction in Avena coleoptiles is studied as a function of the stimulation time. The direction of the stimulation with respect to the vascular bundles must be defined when studying geotropic responses. It is found that the threshold time to evoke geotropic response is less than half a minute, i.e., at least ten times lower than the presentation time usually reported in the literature. An extrapolation procedure can be used to give a so-called extrapolated presentation time t_b, which is intimately related to the logarithmic part of the geotropic response curve and has a physical meaning in the reciprocity rule. The problem of the duration of the true threshold time for stimulation with 1 g is discussed. An experiment indicates that it is not necessary for mass particles (“statoliths”) to settle on the lateral cell wall in order to start the geotropic reaction chain. The slope of the logarithmic part of the geotropic response curve is independent of the transverse force applied to the coleoptiles. Support is given to the view that the slope is determined by the number of sedimenting mass particles.     

Entrainment of Geotropic Oscillations in Hypocotyls of Heliunthus annuus– An Experimental and Theoretical Investigation

A theory for the geotropic movements of the Heliunthus unnuus hypocotyl has been given earlier by Johnsson et ul. In the present paper this theory is tested by extensive experiments. The magnitude as well as the rate of geotropic curvature are found to be in agreement with the theoretical predictions. Further, the geotropic reaction time, defined as the time between the start of geotropic stimulation and the start of the geotropic reaction is determined as a function of the stimulation angle with respect to the plumb line. A slight variation found in the reaction time is included in the theory.     

Reciprocity in the activation of geotropism in oat coleoptiles grown on clinostats

Summary The activation of geotropism in oat coleoptiles, grown on horizonta clinostats, i.e., without the tropic influence of gravity, shows a reciprocal relationship between force and time. Two methods were used to approximate geotropic presentation time. In one, this parameter was estimated by extrapolation to zero response from the linear relationship, response = a+b log stimulation time. In the other, stimulation times of very short as well as longer durations were used; under these conditions, the response curve shows two distinct rates, with the lower rate for stimuli of brief duration. The intersection of the two rate-segments of the response curve was taken as the presentation time. Both methods show reciprocity for the activation of geotropism, but yield significantly different reciprocity constants. The ability of the coleoptile to sense gravity is not affected by gravity compensation. With agar as a growth medium, the magnitude of response to gravity is greater than with sand. However, coleoptiles grown in sand are more sensitive to geotropic activation.Work supported by the U. S. Atomic Energy Commission and the National Aeronautics and Space Administration.     

Versuche zur Analyse der positiven und der negativen geotropischen Reaktionen von Keimwurzeln

Summary In the present work the influence of moist air, of sand and of several solutions on the geotropic behaviour of primary roots is studied. The course of the geotropic movement is the result of a concerted action of positive and negative reactions the intensity and duration of which differ in roots of various species.In pea roots the negative movement appears only during a short stage of development. No direct relation exists between the speed of elongation and the appearance of the negative reaction.Primary roots of Zea mays and of Pisum arvense are indifferent to thigmotropic stimuli. The negative movement has, at least in pea roots a smaller mechanically effective force than the positive movement. Therefore the negative reaction does not appear in sand because it cannot overcome the mechanical resistance of the granular medium.In liquid media pea roots react in another way than in air: here the negative reaction begins later, but it has then more influence on the course of the geotropic curvature.The influence of different cations on the geotropic behaviour and on the elongation of the roots can be understood as a combined action of the osmotic effect and of the specific ionic permeability.In pea roots the negative reaction, which appears during the time from 3 to 6 hours after the induction also depends on a definite level of turgor.Primary roots of Zea mays, which grow in a relatively large angle to the vertical line do not lose their geotropic sensibility. They react like plagiotropic organs.In pea roots relations exist between the development of the positive and the negative reaction and the presence of the cotyledons and the tip of the root.Both reactions are induced at the same time by the gravitional stimulus. Their reaction times, however, are different.The root tip is necessary for the induction of both reactions. The negative curvature also appears when the tip is cut off before the end of the reaction time.The course of the geotropic movement of primary roots is compared with the geotropic behaviour of rhizomes. As a possible explanation of both kinds of reactions a two-hormone-hypothesis is discussed.     

Geoperception in the lentil root cap

Previous analysis showed that, in its initial phase, the geotropic response of Lens culinaris L. roots cannot be explained by a simple action by sliding, pressure or contact of amyloplasts on a sensitive surface located along the longitudinal wall. In this study another mode of action is tested by considering the following parameters as functions of the roots inclination: (1) the distance (d) which the amyloplasts move; (2) their number of contacts (mean c) with parietal cytoplasm; (3) the variable (sin alpha) of the transversal component of the statolith weight (mean M x g sin alpha). It is shown that the initial rate of curvature (mean V), at the various angles, is related to the sedimentation of the amyloplasts by the equation mean V = a log b mean d mean c sin alpha (where a and b are constants). The results obtained prove that the geotropic stimulation is dependent upon the sine of the angle (alpha) of the root inclination and explain the sine rule deviation. The role of statoliths is discussed in the light of recent literature on growth inhibitors which are involved in the geotropic reaction.     

The Geotropic Curvature in Roots of Norway Spruce (Picea abies) Containing Anthocyanins

Seedlings of Norway spruce (Picea abies L.) have been found to synthesize anthocyanins in the root tips as well as in the hypocotyls upon irradiation with white light when kept at 4°C for 6–8 days. In addition, it has also been found that the elongation and the geotropic curvature of spruce roots are dependent on the light conditions. The course of the geotropic curvature in spruce roots containing anthocyanins has been followed during a period of 5 h, in which the seedlings were geotropically stimulated continuously in the horizontal position. When the stimulation was performed in white light and in darkness at 21°C, significantly larger curvatures were observed in the roots pretreated at 4°C in darkness than in the roots containing anthocyanins. The specific curvature (curvature in degrees per mm elongation), however, was approximately the same in both types of roots stimulated in white light. This was due to a retarded elongation of the roots pretreated with light at 4°C and containing anthocyanins. A smaller difference in elongation rate between roots with and without anthocyanins was observed in the dark than in the light, but even in the dark the anthocyanin-containing roots grew more slowly than roots without anthocyanins. In order to find out if it is the anthocyanin content or the illumination which affects the elongation and geotropic curvature in the roots, a series of similar experiments was performed using cress seedlings grown at 4°C in light or darkness. Roots of cress seedlings cultivated under conditions which would induce anthocyanin formation in spruce roots exhibited the highest geotropic responses both in light and darkness as compared to cress seedlings grown at 4°C in darkness. As in the case of spruce roots an increase in elongation was observed in cress roots illuminated during the geotropic stimulation. These similarities in the behaviour made it relevant to compare the development of the geotropic curvature in cress and spruce roots.     

Redistribution of Endogenous Indoleacetic Acid, Abscisic Acid and Gibberellins in Geotropically Stimulated Ribes nigrum Shoots

The presence of IAA, ABA and gibberellins in extracts of shoots of Ribes nigrum was demonstrated by gas-liquid chroma-tography (GLC) for both IAA and ABA and by the lettuce hypocotyl assay for gibberellins. Quantitative estimation of the three substances in extracts from upper and lower halves of shoots which had been kept horizontal, and which showed negative geotropic response after 4 h, indicated a redistribution of hormones during the geotropic stimulation. The ratio of the hormones in lower and upper halves was 3.8:1 and 2.8:1 for IAA and giberellins respectively, whereas the ratio of ABA in upper and lower halves was 2.1:1. There is, however, no evidence for the participation of gibberellins and ABA in the early development of negative geotropic curvatures, since shoots of intact Vicia faba seedlings treated with 100 mg/1 solution of GA₃, ABA and 10 mg/1 IAA for 30 min prior to geotropic stimulation, developed negative geotropic curvatures, although shoots pretreated with 50 and 100 mg/1 IAA did not develop curvature.     

A Theory for Circumnutations in Helianthus annuus

A theory is given for circumnutations in plants, especially hypocotyls of Helianthus annuus, which were used as experimental material The theory is based on the lateral auxin transport, which arises when a gravitational force component acts on the plant. With a suitable time delay between stimulus and response, oscillations or circumnutations should arise. It is possible to describe these oscillation phenomena by the solutions of a differential equation, derived in this paper. The time delay has a central role in this equation. The time delay is assumed to be identical with the geotropic reaction time for the hypocotyls. The ratio between the periodic time for the circumnutations and the reaction time for geotropic curvatures was found to be approximately constant in the temperature region investigated (namely 15–40°C), which supports the theory. Different methods of recording the circumnutations were used, 8 mm film camera technique being the most frequently employed. The introduction of a weighting function for describing the plants “memory” of the stimulation makes it possible to relate the periodic time of the circumnutations to the reaction time for geotropic curvatures. The necessity of this weighting function as well as of the time delay in the equations is emphasized. An explanation of the “Fünfphasen-bewegungen”, reported in the literature, is presented.     

Kinetics of stress relaxation properties of oat coleoptile cell wall after geotropic stimulation

This study describes the stress relaxation of the cell wall of oat (Avena sativa) coleoptiles after different periods of geotropic stimulation. The upper and lower tissues (with respect to gravity) of geotrophically stimulated coleoptiles exhibit different wall properties. The lower tissues are less resistant to deformation than the upper. The ratio of stress to strain is significatly less in the lower than in the upper tissue. Similarly, the relaxation time and the minimum relaxation time, derived from the Maxwell model which describes the physical characteristics of polymers, are also shorter in the lower tissue. However, the maximum relaxation time shows no difference between the upper and lower tissues of a geotropically stimulated coleoptile. The differences between the tissues begin at about 8 minutes after the commencement of stimulation, similar to the time for the initiation of dictyosome redistribution, and precede the onset of geotropism. The above responses of the cell wall of the lower tissue are similar to those induced by indoleacetic acid. The parameters of wall properties of the coleoptiles of both the control and the geostimulated fluctuate rhythmically with time. The periodic changes in wall properties of the coleoptile are compared to other cyclic physiological phenomena.     

The Mechanism of Geoperception in Lentil Roots

The sediment of the amyloplasts in the statocytes of lentilroots was analysed in two different samples of seedlings. Theroots of the first sample (HP) were grown in a horizatal position,whereas thoes of the second sample (VP+20) were grown in a horizontalposition and then exposure to give a response). It is demonstratedthat the statolish can almost enter into contact with the plasmamembrane lining the longitudinal wall of the statocytes of theHP sample. However, these organelles in the VP+20 sample areable to provoke a geotropic stimulation though they are sedimentedat a distance of 0.1350–0.2600 µm from the plasmalemma.From these data it is concluded that the cytoplasmic structureswhich may play a role in the geotropic stimulation are: (1)the endoplasmic reticulum located along the longitudianl wallof the statocytes and (2) the microtubules or the plasmalemmaif the action of the statoliths is transmitted by the parietalcytoplasm. The large aggregations of endoplasmic reticulum whichare situated in the distal part of the central root cap cellswould not have any role in the perception of gravity by roots.These results are discussed in the light of recent work showingthe action of a growth inibitor in the geotropic reaction ofroots.     

The Optimum Angle of Geotropic Stimulation and its Relation to the Starch Statolith Hypothesis

Roots which are turned from their normal direction to directions at various angles with the plumb line develop the largest geotropic curvatures during a subsequent klinostat rotation period when the stimulation angle is well above the horizontal. In experiments with roots of Lepidium sativum L., the optimum is located at 120 to 140° when the stimulation time is between 2 and 15 min. If this fact is to be explained by the movements of amyloplasts in the root cap cells, one would expect roots which bad been kept inverted before the stimulation (so that the moveable amyloplasts are accumulated in the opposite end of the cells) to show an optimum angle well below 90°. — Pre-inversion of the roots did suppress the curvatures produced by stimulation at angles larger than 90° when measured after 10 to 30 min of klinostat rotation. This suppression may be taken as a support for the starch statolith hypothesis, since the amyloplasts in pre-inverted roots placed at angles exceeding 90° have a restricted opportunity to slide along the cell walls compared to non-inverted roots placed at the same angles. In pre-inverted roots measured after a period of klinostat rotation, however, no optimum was found at angles below 90°. When the stimulation time was 3.75 min, the response curves were nearly symmetrical about 90°. Stimulation for 15 min, on the other hand, resulted in curvatures which were much larger (although suppressed in comparison with non-inverted roots) when the stimulation angle was 165° than when it was 15°. During the 15 min stimulation period itself, however, pre-inverted roots curved 0.3° when stimulated at 15, but only 3.4° at 165°. This small difference was very highly significant and is in agreement with the starch statolith hypothesis insofar as the amyloplasts in pre-inverted roots placed at 15° have the greatest opportunity to slide along the cell walls. The lack of further development (and the actual decrease) of their curvatures during the subsequent klinostat rotation must then be due to other, depressing, factors, summarily designated as tonic. At angles above 90°, the tonic factors are either absent or even enhancing. Tbe tonic effects cannot be explained by amyloplast movements.     

Movement of Cell Organelles and the Geotropic Curvature in Roots of Norway Spruce (Picea abies)

The geotropic development in roots of Norway spruce [(Picea abies (L.)] H. Karst, has been followed by light and electron microscopy and compared with the movement of cell organelles (statoliths) in the root cap cells. The geotropic curvature develops in two phases: (a) an initial curvature in the root cap region, which results in an asymmetry in the extreme root tip and which appears after about 3 h stimulation in the horizontal position; and (b) the geotropic curvature in the basal parts of the root tip, which after 8 h is distributed over the entire elongation zone. A graphic extrapolation, based on measurements of the root curvatures after various stimulation periods, indicates a presentation time in the range of 8 to 10 min. The root anatomy and ultrastructure have been examined in detail in order to obtain information as to which organelles may act as gravity receptors. The root cap consists of a central core (columella) distinct from the peripheral part. The core contains three to four rows of parenchymatic cells each consisting of 15 to 18 storeys of statocyte cells with possibly mobile cell organelles. Amyloplasts and nuclei have been found to be mobile in the root cap cells, and the movement of both types of organelles has been followed after inversion of the seedlings and stimulation in the horizontal position for various periods of time at 4°C and 21°C. Three-dimensional reconstructions of spruce root cap cells based on serial sectioning and electron microscopy have been performed. These demonstrate that the endoplasmic reticulum (ER)-system and the vacuoles occupy a considerable part of the statocyte cell. For this reason the space available for free movement of single statolith particles is highly restricted.     

The Physiology of Plant Nutation: I. NUTATION AND GEOTROPIC RESPONSE

The nutation of plant organs has been considered to be eitherthe result of a geotropic feedback loop or produced by an endogenousoscillator. Observations have been made of the angular displacementof Phaseolus seedlings during response to a gravitational stimulus.Nutational oscillations have been observed during the correctivemovement of the stimulated plants in many cases. Geotropic movementcan, however, occur in plants not exhibiting nutation. Theseobservations are considered to support the hypothesis that themovements of nutation in Phaseolus have an endogenous originunconnected with geotropic reactions. A theoretical model isproposed to account for the observed nutational behaviour ofbean seedlings.     

Application of Fine Grain Processing and Condenser Illumination Enlarging to Photomicrography

Photomicrographs involving great resolution are usually made by means of long initial projection. This involves the use of heavy, often cumbersome, apparatus designed to eliminate vibration. This paper evaluates the possibility of using an intermediate projection distance equal to or greater than 160 mm., which is considered the minimum efficient distance, and recording the initial negative image in very fine grain. Then by critical enlarging a positive image is obtained which closely approaches the resolution obtainable by the finest long projection equipment. The initial short projection permits elimination of vibration difficulties attending long exposures so that critical printing at magnifications above 2000 × will give consistently good results, provided fine grain technic is mastered by the operator.     

Entrainment of Geotropic Oscillations in Hypocotyls of Helianthus annuus– An Experimental and Theoretical Investigation

The effects of periodically repeated geotropic stimulations (geotropic pulses) on sunflower hypocotyls are studied. The experimental results agree with predictions from a theory for geotropic movements of the hypocotyl. A sunflower hypocotyl performs circumnutations around or across the plumb line with a period of about 2.5 hours. These oscillations can be entrained to periodically repeated geotropic pulses with a period in a region around 2.5 hours. The limits of this “region of entrainment’ for geotropic pulses are studied. In the region of entrainment resonance curves, i.e., the amplitude of the oscillations plotted as a function of the period of the geotropic pulses, are obtained. The phase differences (measured as time differences) between the geotropic pulse rhythms and the resulting oscillations are also studied. Beat phenomena are predicted and recorded outside the region in which the plants can be entrained. The results are discussed in relation to oscillating phenomena of circadian type (i.e., with a period close to 24 hours), and many similarities are pointed out.     

Ventilation required to entrain small particles from leaves

Particles are blown from leaves when the wind at the height of the particles exceeds a minimum which is about 5 m/sec for some fungal spores. In the moderate winds typical within a canopy of leaves, the minimum is attained at spore height during brief changes in wind or puffs before the boundary layer grows to particle height. The requisite change in speed to remove spores occurs over a sizeable area only when the speed changes abruptly in a short distance in the direction of the wind.     

Cell elongation in the grass pulvinus in response to geotropic stimulation and auxin application

Summary Horizontally-placed segments of Avena sativa L. shoots show a negative geotropic response after a period of 30 min. This response is based on cell elongation on the lower side of the leaf-sheath base (pulvinus). Triticum aestivum L., Hordeum vulgare L. and Secale cereale L. also show geotropic responses that are similar to those in Avena shoots. The pulvinus is a highly specialized organ with radial symmetry and is made up of epidermal, vascular, parenchymatous and collenchymatous tissues. Statoliths, which are confined to parenchyma cells around the vascular bundles, sediment towards the gravitational field within 10–15 min of geotropic stimulation. Collenchymatous cells occur as prominent bundle caps, and in Avena, they occupy about 30% of the volume of the pulvinus. Geotropic stimulation causes a 3- to 5-fold increase in the length of the cells on the side nearest to the center of the gravitational field. Growth can also be initiated in vertically-held pulvini by the application of indole-3-acetic acid, 1-naphthaleneacetic acid or 2.4-dichlorophenoxyacetic acid. 2.3.5.-triiodobenzoic acid interferes with growth response produced by geotropic stimulation as well as with the response caused by auxin application. Gibberellic acid and kinetin have no visible effect on the growth of the pulvinus. Polarization microscopy shows a unique, non-uniform stretching of the elongating collenchymatous cells. Nonelongated collenchymatous cells appear uniformally anisotropic. After geotropic stimulation or auxin application, they appear alternately anisotropic and almost isotropic. Such a pattern of cell elongation is also observed in collenchyma cells of geotropically-stimulated shoots of Rumex acetosa L., a dicotyledon.Abbreviations 2.4-D 2.4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA l-naphthaleneacetic acid - TIBA 2.3.5-triiodobenzoic acid     

The Effect of Light on the Geotropic Responses of Phycomyces Sporangiophores

The geotropic responses of Phycomyces sporangiophores were studied under varying intensities of illumination, using a low speed centrifuge and a fixed beam of blue light. This light has a strongly inhibitory effect on the transient geotropic response, reducing it to 36 per cent of its magnitude in darkness. The inhibition does not vary systematically with light intensity over a range of 400-fold. The light sensitivity of the transient geotropic response thus differs from the light-growth response system, which shows the same growth rate in light and darkness. By contrast, the slower long term geotropic response is enhanced by light of moderate intensities, but is strongly inhibited by high intensities. At and above a mean intensity of about 1 µw/cm², the long term response is completely removed. If the intensity is lowered from an inhibitory level, either to darkness or to a low level, the geotropic response appears after a time lag of 20 minutes. Furthermore an increase in intensity from one level to another, both levels normally enhancing, results in a transient reversal in the long term geotropic response, also after a time lag of 20 minutes. Thus it is suggested that light is acting at some intermediate step in the long term geotropic sensory system, a step that normally requires 20 minutes for completion.     

The Change of Tropism in Polyporus brumalis Stipes and the Effect of Directional Stimuli on Pileus Differentiation

The directional influences of light and gravity upon the developmentof pileate and epileate sporophores of Polyporus brumalis, producedin pure culture, have been investigated. Growth in the dimiticfruit-body has a considerable subapical component which in thestipe is responsible for tropistic curvature. The stipe is competentto react negatively to unilateral gravitational and positivelyto unilateral photic stimulation throughout development, butwhen both stimulioperate phototropism masks geotropism. If illuminatedfrom one side the growing epileate stipe is strongly positivelyphototropic but as the pileus reaches a characteristic diameterthe stipe becomes negatively geotropic. Experiments with changingdirection of illumination and with artificial pilei of blackpaper suggest that the change of tropism is explicable by theshading action of the expanding pileus on the sub-pilcal photoperceptiveand photoreactive region of the stipe. Developing sporophores continuously rotated with reference tofixed directions of gravitational and light stimulation andothers with stipes inverted as a result of illumination frombelow during development have in common that the morphologicallyupper surface of the pileus always develops towards and approximatelyat right angles to the direction from which the maximum lightintensity is received. Normal but inverted stipes and dissepimentsshow no tendency to geotropic reorientation.     

Studies on the Geotropism of Roots: I.GROWTH-RATE DISTRIBUTION DURING RESPONSE AND THE EFFECTS OF APPLIED AUXINSI

Experiments are described in which the normal geotropic responsesof the roots of Pisum sativum seedlings have been compared withthose obtained in the presence of auxins (indole-3-acetic acidand 2:4-dichlorophenoxyacetic acid) in the external medium.The courses of positive curvature resulting from short exposures(40 minutes) and also subsequent recovery phenomena on a horizontalklinostat have been followed. A photographic recording techniqueallowed the determination of absolute growth-rates of both upperand lower sides of the root during the course of each experiment. Positive curvature started at its maximum rate (0.30–0.32deg./min.) after a reaction time of 11.5 minutes and continuedconstant at that rate for about 60 minutes after stimulationceased. Recovery took place at a similar rate of curvature andwas complete after a further 150–200 minutes. During thephase of positive curvature overall root growth-rates were considerablyreduced and were slowly restored to normal during recovery. Low concentrations (1 part in 10¹¹) of both auxins increasedthe rate of positive curvature by 30–40 per cent. andshortened the reaction time roughly in proportion. The growth-ratesof both sides of the root were increased to the same extentduring both curvature and recovery. High concentrations (10^–8IAA and 3.10^–8 2: 4-D)reduced the rates of curvature by 50 per cent., lengthened thereaction time, and inhibited the growth of both sides of theroot during both curvature and recovery. Neither concentration of either auxin otherwise affected thetime course of response and recovery. It is suggested that geotropic response is due to the de novoproduction of an endogenous inhibitor in the extending cellsof the lower side of the root whence it may later spread tothe upper side. The complete independence of the growth actionsof this inhibitor and of the applied auxins suggests that itis not indole-3-acetic acid or any similar compound. Recoverymay be very largely independent of both inhibitor and auxinsand due to the action of another growth factor limiting celllength. The implications of these findings and of the attendant theoriesare fully discussed.