A digital sensor to measure real-time leaf movements and detect abiotic stress in plants

Plant and plant organ movements are the result of a complex integration of endogenous growth and developmental responses, partially controlled by the circadian clock, and external environmental cues. Monitoring of plant motion is typically done by image-based phenotyping techniques with the aid of computer vision algorithms. Here we present a method to measure leaf movements using a digital inertial measurement unit (IMU) sensor. The lightweight sensor is easily attachable to a leaf or plant organ and records angular traits in real-time for two dimensions (pitch and roll) with high resolution (measured sensor oscillations of 0.36 ± 0.53° for pitch and 0.50 ± 0.65° for roll). We were able to record simple movements such as petiole bending, as well as complex lamina motions, in several crops, ranging from tomato to banana. We also assessed growth responses in terms of lettuce rosette expansion and maize seedling stem movements. The IMU sensors are capable of detecting small changes of nutations (i.e. bending movements) in leaves of different ages and in different plant species. In addition, the sensor system can also monitor stress-induced leaf movements. We observed that unfavorable environmental conditions evoke certain leaf movements, such as drastic epinastic responses, as well as subtle fading of the amplitude of nutations. In summary, the presented digital sensor system enables continuous detection of a variety of leaf motions with high precision, and is a low-cost tool in the field of plant phenotyping, with potential applications in early stress detection.

An inertial measurement unit is capable of measuring dynamic and complex plant organ movements in real-time, and is suitable for early abiotic stress detection.     

An automatic device to record the force necessary to compensate plant movements

A device, which automatically and continuously keeps moving plants in fixed positions, has been constructed. The apparatus is based on optical detection of any movements of the plant organ under study. The plant organ is kept in the desired position by means of wires, and the force necessary to apply to the wires to achieve this is recorded. The force reflects any tendency of the plant organ to move. The system is controlled by an Apple II computer. The device can work in one or two dimensions and record compensation forces in a wide range. In the experiments mentioned below, the magnitude of the forces necessary to keep the plants in a fixed position was of the order of 10^-4N.
Circadian leaf movements of Oxalis regnellii Mig. were studied in the device. The leaf rhythm continued, although the leaflets were clamped and the light input on the leaf therefore constant. Circumnutation of hypocotyls of Helianthus annus L. cv. Californicus were drastically reduced in amplitude when the hypocotyls were kept in vertical position by the wires. Since the gravitropic input signal to this system was zeroed by the equipment, the results demonstrated that in the absence of gravitropic inputs the circumnutation reactions drastically diminish. This confirmed that circumnutations of these hypocotyls are influenced by gravity. Finally, the apparatus was used to study phototropic reactions: By clamping phototropically stimulated coleoptiles of Avena sativa L. cv. Seger no gravitational stimulations were involved and the phototropic reaction without interference from gravity could be studied.     

A simple and inexpensive system for monitoring jaw movements in ambulatory humans

A simple and inexpensive method for recording vertical movements of the human mandible relative to the maxilla is presented. Measurements are made from accelerometers and a Hall-effect device temporarily glued to the upper and lower anterior teeth. The accelerometer signals are integrated once to give velocity and a second time to give position. Movements of the mandible relative to the maxilla are obtained by integrating the difference between the two accelerometer signals. The (relative) velocity and position records derived in this way are linear, but subject to drift when the jaw is stationary. Steady mandibular position is obtained from the Hall-effect system, but this signal must be corrected for its inherent non-linearity. This device can record rapid movements of the mandible even when the head is unrestrained, and interferes minimally with normal jaw movements.     

Comparative analysis of diadochokinetic movements.

Tests of diadochokinesia are an inherent part of a neurological examination. Various quantifying methods have been proposed to increase the objectivity, sensitivity, and reliability of such examinations. The methods used and analyses performed, however, differ substantially between tasks. We used a three-dimensional, ultrasound-based recording device to continuously record joint angles during three diadochokinetic movements, avoiding any external constraints of the movements. Alternate pronation and supination of the forearm, tapping with the whole hand and with the index finger in isolation were analyzed in a sample of 63 healthy control subjects. The most sensitive measure for capturing effects of gender, sex, and active hand was frequency. The right hand was faster than the left in all tasks, tapping performance declined with increasing age, and male subjects were faster than females in forearm diadochokinesia. Other measures that characterize speed of movement such as maximum angular velocities and accelerations did not yield comparable sensitivity in detecting the same statistical effects. However, angular velocity achieved the highest test-retest reliability for forearm diadochokinesia, while frequency was reproduced in the tapping tasks. Additional measures characterizing symmetry of the angular velocity profiles and intraindividual variability were shown to be largely independent of movement speed. Examples in neurological patients showed that the data define a valuable standard against which pathological performance can be precisely evaluated. In addition, the different measures captured dissociable aspects of motor performance that may further help to characterize the deficit and adjust therapy.     

Assessment of inertial and gravitational inputs to the vestibular system

A new device for the assessment of instantaneous angular and linear accelerations of the head is presented, which is based on four linear tri-axial accelerometers suitably attached to the head by an helmet. A procedure for reproducible helmet placement and calibration is given. A method is also illustrated to work out the different linear accelerations sensed by the vestibular organs in the left and right labyrinths and the components of the angular acceleration sensed by their semicircular canals. The computation is based on few individual parameters describing the helmet position with respect to external landmarks and on the average internal position and orientation of the vestibula. The purpose is to study the components of internal inertial forces, which represent the primary inputs to the vestibular system devoted to equilibrium and oculomotor control. The system is designed to be of easy application during rehabilitation exercises and in clinical environment during diagnostic and therapeutic manoeuvres. The prototype is tested with simple free movements such as "yes", "no", and gait.     

Analysis of the middle latency evoked potentials to angular acceleration impulses in man

The middle latency vestibular evoked potential (ML-VsEP) recorded with scalp electrodes in man in response to impulses of angular acceleration is dominated by a forehead positive peak at about 15 ms and a negative peak at about 20 ms; the peak amplitude of this component is about 30 μV. This is followed by slower, smaller amplitude activity. The latency of this initial peak is similar to the latency of the vestibulo-ocular reflex (VOR) in monkeys. The present study was undertaken to elucidate the possible relation between the ML-VsEPs and VOR. This included recordings from forehead-mastoid electrodes (sites used to record VsEP) and other scalp electrodes and the recording of potentials due to eye movement: the electro-oculogram. Direct recording of eye movements was also conducted using an infra-red reflection device in those experiments in which the head was not moved. The recordings were conducted in man during vestibular stimulation eliciting VsEPs, during voluntary eye movements and during caloric and optokinetic stimulation. These experiments indicated that the 15–20 ms component of the ML-VsEP was not due to movements of the eye (corneoretinal dipole). The large amplitude 15–20 ms component of the ML-VsEP was similar in general magnitude, waveform, polarity, duration and rise time to the highly synchronous pre-saccadic spike (neural and/or myogenic) which precedes nystagnys and voluntary saccades. It therefore probably represents vestibular-initiated electrical activity in motor units of the extra-ocular muscles which then produce anti-compensatory saccades.     

SUN-TRACKING AND RELATED LEAF MOVEMENTS IN A DESERT LUPINE (LUPINUS ARIZONICUS)

Both diaheliotropic (sun-tracking) and paraheliotropic (cupping) leaf movements are described for the Arizona Lupine [Lupinus arizonicus (Wats.) Wats.]. The leaf movements are shown to be non-circadian in nature. Evidence is presented that an active K⁺ ion transport mechanism is involved in these turgor-related leaf movements. Increasing concentrations of lanthanum, a known ion transport inhibitor, showed increasing inhibition of both leaf movements. Increasing concentrations of other salts did not inhibit either leaf movements, instead there was an increase in the cupping leaf movement (elevation of the leaflets) which is shown to be a water stress response.     

The simulation of aerial movement--III. The determination of the angular momentum of the human body

A method is presented for determining the angular momentum of the human body about its mass centre for general three-dimensional movements. The body is modelled as an 11 segment link system with 17 rotational degrees of freedom and the angular momentum of the body is derived as a sum of 12 terms, each of which is a vector function of just one angular velocity. This partitioning of the angular momentum vector gives the contribution due to the relative segmental movement at each joint rather than the usual contribution of each segment. A method of normalizing the angular momentum is introduced to enable the comparison of rotational movements which have different flight times and are performed by athletes with differing inertia parameters. Angular momentum estimates were calculated during the flight phases of nine twisting somersaults performed on trampoline. Errors in film digitization made large contributions to the angular momentum error estimates. For individual angular momentum estimates the relative error is estimated to be about 10% whereas for mean angular momentum estimates the relative error is estimated to be about 1%.     

Differentially Phased Leaf Growth and Movements in Arabidopsis Depend on Coordinated Circadian and Light Regulation

In contrast to vastly studied hypocotyl growth, little is known about diel regulation of leaf growth and its coordination with movements such as changes in leaf elevation angle (hyponasty). We developed a 3D live-leaf growth analysis system enabling simultaneous monitoring of growth and movements. Leaf growth is maximal several hours after dawn, requires light, and is regulated by daylength, suggesting coupling between growth and metabolism. We identify both blade and petiole positioning as important components of leaf movements in Arabidopsis thaliana and reveal a temporal delay between growth and movements. In hypocotyls, the combination of circadian expression of PHYTOCHROME INTERACTING FACTOR4 (PIF4) and PIF5 and their light-regulated protein stability drives rhythmic hypocotyl elongation with peak growth at dawn. We find that PIF4 and PIF5 are not essential to sustain rhythmic leaf growth but influence their amplitude. Furthermore, EARLY FLOWERING3, a member of the evening complex (EC), is required to maintain the correct phase between growth and movement. Our study shows that the mechanisms underlying rhythmic hypocotyl and leaf growth differ. Moreover, we reveal the temporal relationship between leaf elongation and movements and demonstrate the importance of the EC for the coordination of these phenotypic traits.     

Critical illness VR rehabilitation device (X-VR-D): Evaluation of the potential use for early clinical rehabilitation

We present a new critical illness VR rehabilitation device (X-VR-D) that enables diversified self-training and is applicable early in the rehabilitation of severely injured or ill patients. The X-VR-D consists of a VR program delivering a virtual scene on a flat screen and simultaneously processing commands to a moving chair mounted on a motion system. Sitting in the moving chair and exposed to a virtual reality environment the device evokes anticipatory and reactive muscle contractions in trunk and extremities for postural control. In this study we tested the device in 10 healthy subjects to evaluate whether the enforced perturbations indeed evoke sufficient and reproducible EMG muscle activations.

We found that particular fast roll and pitch movements evoke adequate trunk and leg muscle activity. Higher angular velocities and higher angles of inclination elicited broader EMG bursts and larger amplitudes. The muscle activation pattern was highly consistent between different subjects and although we found some habituation of EMG responses in consecutive training sessions, the general pattern was maintained and was predictable for specific movements. The habituation was characterized by more efficient muscle contractions and better muscle relaxation during the rest positions of the device. Furthermore we found that the addition of a virtual environment to the training session evoked more preparatory and anticipatory muscle activation than sessions without a virtual environment.

We conclude that the X-VR-D is safe and effective to elicit consistent and reproducible muscle activity in trunk and leg muscles in healthy subjects and thus can be used as a training method.     

In situ Observations of Stomatal Movements

Kappen, L., Andresen, G. and L?sch, R. 1987. In situ observationsof stomatal movements.—J. exp. Bot. 38: 126–141. A device is described by which stomatal movements in situ canbe observed and recorded continuously in light and in darkness.It is mounted in a conditioned CO₂ exchange measuring chamberso that stomatal movements can be observed whilst CO₂ exchange(photosynthesis and respiration) of the same leaf is measured.Advantages and limitations are discussed. By this method itwas shown that stomata of Vicia faba although responding inthe same direction to environmental stimuli exhibited a widerange of pore widths. Responses to changes of air humidity andof CO₂ content were clearly evident when the leaves were exposedto light. Before stomata closed due to decreasing water vapourpressure differences between leaf and air they showed a markedwidening of the pore. An inverse response occurred when watervapour pressure deficit decreased. In darkness stomata did notrespond to such changes. Key words: Stomata, leaf gas exchange, microscopic observation     

Anatomical and physiological characteristics of the petiole of Abutilon theophrasti in relation to circadian leaf movements

Leaf movements in Abutilon theophrasti Medic, were monitored manually and by a continuous electronic recording device. Plants entrained to a daily regime of a 15 h light span followed by 9 h of darkness showed rhythmic movements that persisted under conditions of continuous illumination and constant temperature with a circadian period. The rhythmic change in orientation of the leaf from a near horizontal (day) to a near vertical (night) position was attributed to movement of the blade and not the petiole. The end of the petiole next to the blade functions as a joint or pulvinus. Anatomical confirmation of the existence of a pulvinus in the Abutilon leaf was provided by light microscopy. Vascular tissue in this region forms a solid cylinder with no pith, and the cortex is parenchymatous. In the main part of the petiole, the vascular tissue is arranged in four to six bundles, a pith is present, and the cortex contains a sub-epidermal ring of collenchyma. Both the functional and anatomical evidence indicate the presence of a pulvinus that functions in circadian leaf movements of Abutilon.     

Stomatal responses to changes in atmospheric humidity and water supply: Experiments with leaf sections of Zea mays in CO2-free air

Summary Leaf sections were exposed to CO₂-free air, thus excluding interference by the CO₂-sensitive system in the guard cells. Stomates did not close in response to change from moist to dry air, whether it passed over the leaf or was forced through the intercelluar spaces. In contrast, the stomatal apertures became narrower if the water potential in the liquid supplying the leaf was lowered. Of solutions with the same osmolality, those with the higher viscosity produced the larger responses.Transient stomatal movements in the opposite direction to the final response were observed upon any sudden change in the water status of the leaf sections, whether caused by varying the moisture content of the air passing around or through the leaf sections, or by varying the water supply. Increased load on the water supply caused temporary opening movements, while improvements in water supply led to closing movements of varying duration. When dry air was forced through the leaf sections, non-sinusoidal oscillations with large amplitudes were sometimes observed.It is concluded that the guard cells are tightly coupled to the water-supply system of the leaf and only indirectly to the conditions in the atmosphere by a negative feedback of transpiration on the water potential in the water-conducting system.     

The effect of lunisolar tidal acceleration on stem elongation growth,nutations and leaf movements in peppermint (Mentha × piperita L.)

• Orbital movement of the Moon generates a system of gravitational fields that periodically alter the gravitational force on Earth. This lunar tidal acceleration (Etide) is known to act as an external environmental factor affecting many growth and developmental phenomena in plants. Our study focused on the lunar tidal influence on stem elongation growth, nutations and leaf movements of peppermint.
• Plants were continuously recorded with time‐lapse photography under constant illumination as well in constant illumination following 5 days of alternating dark–light cycles. Time courses of shoot movements were correlated with contemporaneous time courses of the Etide estimates. Optical microscopy and SEM were used in anatomical studies.
• All plant shoot movements were synchronised with changes in the lunisolar acceleration. Using a periodogram, wavelet analysis and local correlation index, a convergence was found between the rhythms of lunisolar acceleration and the rhythms of shoot growth. Also observed were cyclical changes in the direction of rotation of stem apices when gravitational dynamics were at their greatest. After contrasting dark–light cycle experiments, nutational rhythms converged to an identical phase relationship with the Etide and almost immediately their renewed movements commenced. Amplitudes of leaf movements decreased during leaf growth up to the stage when the leaf was fully developed; the periodicity of leaf movements correlated with the Etide rhythms.
• For the fist time, it was documented that lunisolar acceleration is an independent rhythmic environmental signal capable of influencing the dynamics of plant stem elongation. This phenomenon is synchronised with the known effects of Etide on nutations and leaf movements.

     

Thermonastic leaf movements: a synthesis of research with Rhododendron

Thermonastic leaf movements in Rhododendron L. occur in response to freezing temperatures. These movements are composed of leaf curling and leaf angle changes that are distinct leaf movements with different responses to climatic factors. Leaf angle is controlled by the hydration of the petiole, as affected by soil water content, atmospheric vapour pressure, and air temperature. In contrast, leaf curling is a specific response to leaf temperature, and bulk leaf hydration has little effect. The physiological cause of leaf curling is not well understood, but the mechanism must lie in the physiology of the cell wall and/or regional changes in tissue hydration. Available evidence suggests that intercellular freezing is not a cause of leaf curling.
Manipulation experiments demonstrate that changes in leaf orientation in Rhododendron most likely serve to protect the leaves from membrane damage due to high irradiance and cold temperatures. In particular, the pendent leaves protect the chloroplast from photoinhibition. Leaf curling may serve to slow the rate of thaw following freezing, a common phenomenon in the Appalachian mountains of the U.S. The thermonastic leaf movements have a greater importance to plants in a dim environment because the potential impact to canopy carbon gain is greater than in high light environments.
These leaf movements have several implications for horticultural management. There seems to be a trade-off between water stress tolerance and freezing stress tolerance by leaf movements. Thermonastic leaf movements may be a major mechanism of cold stress tolerance in Rhododendron species. The actual physiological cause of leaf movement has not been elucidated and many more species need to be evaluated to verify the general importance of leaf movements to Rhododendron ecology and evolution.     

Heliotropic leaf movements in common beans controlled by air temperature

Heliotropic leaf movements were examined in common beans (Phaseolus vulgaris cv Blue Lake Bush) under outdoor and laboratory conditions. Heliotropic leaf movements in well-watered plants were partly controlled by temperature, and appeared to be independent of atmospheric humidity and CO₂ concentration. When environmental conditions were held constant in the laboratory, increased air temperature caused bean leaves to orient more obliquely to a light source. Ambient CO₂, intercellular CO₂, and net photosynthesis were not correlated with the temperature-induced changes in heliotropic movements, nor did they significantly affect these movements directly. The effect of air temperature on leaf movements need not be mediated through a change in leaf water potential, transpiration, or leaf conductance. Air temperature modified laminar orientation in light through its effect on tissue temperature in the pulvinal region, not that of the lamina or petiole. However, under darkness the temperature effects on leaf movements were not expressed. Active heliotropic movements in response to air temperature allowed lamina temperature to remain close to the thermal optimum of photosynthesis. This temperature effect underlies a commonly observed pattern of leaf movements under well-watered conditions: a tendency for leaves to face the sun more obliquely on hot days than cool days.     

Concurrent validity and reliability of a mobile tracking technology to measure angular and linear movements of the neck

The neck can be moved in six degrees of freedom. Current 3D-optoelectronic motion-capture systems capable of measuring these movements are inappropriate for use in clinical practice because they are stationary, expensive and time-consuming. We therefore developed a less complex 3D-tracking technology based on Steam®VR to measure six degrees of freedom in a clinical setting. The aim of this study was to assess the validity and reliability of this system.The developed prototype consists of two infrared-emitting lighthouses and sensors, mounted on the participant’s helmet and trunk belt, to detect the orientation of the head and trunk. The system was evaluated by means of an infrared light-reflecting marker tracking system. Twenty healthy participants, equipped with these sensors and markers, performed thirteen neck movement tasks. Linear and angular movements were measured. These tasks were repeated after six to eight days to assess test-retest reliability. Concurrent validity was assessed by the root mean square error, and reliability with generalizability theory.With an average root mean square error between 1.2 and 2.0° in angular and 0.4–0.5 cm in linear movements, the prototype was shown to precisely track these movements. Reliability of the prototype and the reference system was comparable for all tasks. A high contribution of participant’s variability to the observed variance was generally detected, with the exception of joint repositioning error and upper cervical flexion.The reliability was task-specific and did not differ between the systems. The prototype system was shown to be valid, although the reliability of the repositioning and upper cervical flexion tests needs to be reconsidered.     

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.     

Response of the ArcCHECK® device at 6 MV and 15 MV for VMAT and IMRT quality control

PurposeTo study the response of the ArcCHECK® device as VMAT and IMRT verification system.MethodsVarious tests analyzing the linearity, the repeatability and the angular dependence of the device response, its dependence with the pulse repetition rate and the leakage losses were performed. The long-term response in dose measurements and the uniformity of the detectors conforming the system were controlled using a statistical process control program. The Elekta Infinity™ 6 and 15 MV photon beams were used.ResultsThe device showed excellent repeatability and linearity. The differences between the responses obtained for any pair of angular incidences were less than 2%. The absorbed dose increased by 3% when the pulse repetition rate varied from 50 to 600 MU/min. Results are in overall agreement with those found in previous works for the ArcCHECK®, in which a reduced number of the device diodes were analyzed, and for the MapCheck®, an older 2D device that used the same diodes. Charge losses were found to be negligible except for some of the diodes of the device. The statistical process control program is a very useful tool to control the correct functioning of the device in the long term.ConclusionsThe results of the analysis carried out indicate that the working and stability conditions of the ArcCHECK® device are adequate for its purpose. The dependence with the pulse repetition rate should be considered when VMAT or similar treatments are evaluated. A control program for the statistical monitoring of the device would be desirable and useful.     

Basic properties of the circadian leaf movements of Oxalis regnellii, and period change due to lithium ions

The circadian leaf movement of Oxalis regnellii Mig, has been investigated. The three leaflets of a stalk were normally synchronized, and under the experimental conditions chosen they showed a period of 26.2 ± 0.1 h. Cutting off one or two leaflets led to a successive decrease of the period length (25.7 ± 0.1 and 25.1 ± 0.3 h resp.). It was possible to phase shift the leaf movements by mechanical means (advance of 1.6 ± 0.3 h).
Lithium ions, added permanently to the transpiration stream, increased the period length of the leaf movements by more than one hour (with 10 m M Li⁺). A 24 h pulse of 20 m M LiCl caused a permanent 2–3 h phase delay of the circadian rhythm. Four-h pulses, on the other hand, provoked only transient phase delays, the magnitude being dependent on the phase of application. Lithium concentrations were determined for different regions of leaves and pulvini. It was shown that leaf segments had considerably lower concentrations than pulvini. No significant difference in the lithium concentration was observed between the upper and lower part of pulvini.
In the light leaf position was strongly correlated with water uptake and the consequences for applications of substances to the circadian system via the transpiration stream is discussed. A simple model of the oscillatory system and reactions connected to it is discussed.