The shaping of plant axes and crowns through tropisms and elasticity: an example of morphogenetic plasticity beyond the shoot apical meristem.

Shoot morphogenetic plasticity is crucial to the adaptation of plants to their fluctuating environments. Major insights into shoot morphogenesis have been compiled studying meristems, especially the shoot apical meristem (SAM), through a methodological effort in multiscale systems biology and biophysics. However, morphogenesis at the SAM is robust to environmental changes. Plasticity emerges later on during post-SAM development. The purpose of this review is to show that multiscale systems biology and biophysics is insightful for the shaping of the whole plant as well. More specifically, we review the shaping of axes and crowns through tropisms and elasticity, combining the recent advances in morphogenetic control using physical cues and by genes. We focus mostly on land angiosperms, but with growth habits ranging from small herbs to big trees. We show that generic (universal) morphogenetic processes have been identified, revealing feedforward and feedback effects of global shape on the local morphogenetic process. In parallel, major advances have been made in the analysis of the major genes involved in shaping axes and crowns, revealing conserved genic networks among angiosperms. Then, we show that these two approaches are now starting to converge, revealing exciting perspectives.

Refinement of international recommendations for cubicles, based on the identification of associations between cubicle characteristics and dairy cow welfare measures.

Maladjusted cubicles for dairy cattle may cause increased skin alterations, lameness, and dirtiness. The International Commission of Agricultural and Biosystems Engineering has produced several recommendations for cubicle design, but a previous study showed that not all of them seem efficient. Here, we aim to refine and complete these recommendations. We collected data on 76 dairy farms (2,404 cows). We modeled the association between combinations of cubicle properties (e.g., type of bedding litter) and dimensions (e.g., cubicle width) relative to cow size, and prevalence of cow skin alterations, lameness, and dirtiness. We used weighted multivariable logistic regression models to predict the presence of skin alteration on the carpus; the neck, shoulder, and back; the flank, side, and udder; and the tarsus or hindquarters. We also evaluated the presence of lameness as well as the dirtiness of the lower hind legs including hocks; the hindquarters, upper hind legs, and flank; the cow rear including tail; and the udder. The risk factors highlighted led us to recommend (1) position cubicles in a way that leaves more than 1 m of clearance from any obstacle in front of the cubicle; (2) if there is an obstacle on the lateral plane (i.e., where the cubicle partition is) in front ahead of the cow, put the obstacle in front of the fore knees; (3) if there is an obstacle in front of the cow on the median plane (e.g., neck or front rail), the position the obstacle between 1.25 and 1.5 of the cow length from the curb and between 1.0 and 1.25 of its height; (4) use curb height between 0.11 and 0.15 of cow height with no sharp edges on the curb; (5) use round or at least has no sharp edges brisket board; (6) use a stone-free soil instead of concrete or use a mattress thicker than 1 cm, with microrelief, and a soft fixing area at the curb, (7) litter with straw (rather than nothing or sawdust) and keep it dry. This risk factor analysis should be followed by experiments in controlled environments to further validate these conclusions and used to update the International Commission of Agricultural and Biosystems Engineering recommendations.

Virtual reality for freely moving animals.

Standard animal behavior paradigms incompletely mimic nature and thus limit our understanding of behavior and brain function. Virtual reality (VR) can help, but it poses challenges. Typical VR systems require movement restrictions but disrupt sensorimotor experience, causing neuronal and behavioral alterations. We report the development of FreemoVR, a VR system for freely moving animals. We validate immersive VR for mice, flies, and zebrafish. FreemoVR allows instant, disruption-free environmental reconfigurations and interactions between real organisms and computer-controlled agents. Using the FreemoVR platform, we established a height-aversion assay in mice and studied visuomotor effects in Drosophila and zebrafish. Furthermore, by photorealistically mimicking zebrafish we discovered that effective social influence depends on a prospective leader balancing its internally preferred directional choice with social interaction. FreemoVR technology facilitates detailed investigations into neural function and behavior through the precise manipulation of sensorimotor feedback loops in unrestrained animals.

Posture control in land plants: growth, position sensing, proprioception, balance, and elasticity.

The colonization of the atmosphere by land plants was a major evolutionary step. The mechanisms that allow for vertical growth through air and the establishment and control of a stable erect habit are just starting to be understood. A key mechanism was found to be continuous posture control to counterbalance the mechanical and developmental challenges of maintaining a growing upright structure. An interdisciplinary systems biology approach was invaluable in understanding the underlying principles and in designing pertinent experiments. Since this discovery previously held views of gravitropic perception had to be reexamined and this has led to the description of proprioception in plants. In this review, we take a purposefully pedagogical approach to present the dynamics involved from the cellular to whole-plant level. We show how the textbook model of how plants sense gravitational force has been replaced by a model of position sensing, a clinometer mechanism that involves both passive avalanches and active motion of statoliths, granular starch-filled plastids, in statocytes. Moreover, there is a transmission of information between statocytes and other specialized cells that sense the degree of organ curvature and reset asymmetric growth to straighten and realign the structure. We give an overview of how plants have used the interplay of active posture control and elastic sagging to generate a whole range of spatial displays during their life cycles. Finally, a position-integrating mechanism has been discovered that prevents directional plant growth from being disrupted by wind-induced oscillations.

Quantitative dissection of variations in root growth rate: a matter of cell proliferation or of cell expansion?

Plant organ growth results from cell production and cell expansion. Deciphering the contribution of each of these processes to growth rate is an important issue in developmental biology. Here, we investigated the cellular processes governing root elongation rate, considering two sources of variation: genotype and disturbance by chemicals (NaCl, polyethylene glycol, H2O2, abscisic acid). Exploiting the adventitious rooting capacity of the Populus genus, and using time-lapse imaging under infrared-light, particle image velocimetry, histological analysis, and kinematics, we quantified the cellular processes involved in root growth variation, and analysed the covariation patterns between growth parameters. The rate of cell production by the root apical meristem and the number of dividing cells were estimated in vivo without destructive measurement. We found that the rate of cell division contributed more to the variation in cell production rate than the number of dividing cells. Regardless of the source of variation, the length of the elongation zone was the best proxy for growth rate, summarizing rates of cell production and cell elongation into a single parameter. Our results demonstrate that cell production rate is the main driver of growth rate, whereas elemental elongation rate is a key driver of short-term growth adjustments.

KymoRod: a method for automated kinematic analysis of rod-shaped plant organs.

A major challenge in plant systems biology is the development of robust, predictive multiscale models for organ growth. In this context it is important to bridge the gap between the, rather well-documented molecular scale and the organ scale by providing quantitative methods to study within-organ growth patterns. Here, we describe a simple method for the analysis of the evolution of growth patterns within rod-shaped organs that does not require adding markers at the organ surface. The method allows for the simultaneous analysis of root and hypocotyl growth, provides spatio-temporal information on curvature, growth anisotropy and relative elemental growth rate and can cope with complex organ movements. We demonstrate the performance of the method by documenting previously unsuspected complex growth patterns within the growing hypocotyl of the model species Arabidopsis thaliana during normal growth, after treatment with a growth-inhibiting drug or in a mechano-sensing mutant. The method is freely available as an intuitive and user-friendly Matlab application called KymoRod.

The Kinematics of Plant Nutation Reveals a Simple Relation between Curvature and the Orientation of Differential Growth.

Nutation is an oscillatory movement that plants display during their development. Despite its ubiquity among plants movements, the relation between the observed movement and the underlying biological mechanisms remains unclear. Here we show that the kinematics of the full organ in 3D give a simple picture of plant nutation, where the orientation of the curvature along the main axis of the organ aligns with the direction of maximal differential growth. Within this framework we reexamine the validity of widely used experimental measurements of the apical tip as markers of growth dynamics. We show that though this relation is correct under certain conditions, it does not generally hold, and is not sufficient to uncover the specific role of each mechanism. As an example we re-interpret previously measured experimental observations using our model.

A unified model of shoot tropism in plants: photo-, gravi- and Propio-ception.

Land plants rely mainly on gravitropism and phototropism to control their posture and spatial orientation. In natural conditions, these two major tropisms act concurrently to create a photogravitropic equilibrium in the responsive organ. Recently, a parsimonious model was developed that accurately predicted the complete gravitropic and proprioceptive control over the movement of different organs in different species in response to gravitational stimuli. Here we show that the framework of this unifying graviproprioceptive model can be readily extended to include phototropism. The interaction between gravitropism and phototropism results in an alignment of the apical part of the organ toward a photogravitropic set-point angle. This angle is determined by a combination of the two directional stimuli, gravity and light, weighted by the ratio between the gravi- and photo-sensitivities of the plant organ. In the model, two dimensionless numbers, the graviproprioceptive number B and the photograviceptive number M, control the dynamics and the shapes of the movement. The extended model agrees well with two sets of detailed quantitative data on photogravitropic equilibrium in oat coleoptiles. It is demonstrated that the influence of light intensity I can be included in the model in a power-law-dependent relationship M(I). The numbers B and M and the related photograviceptive number D are all quantitative genetic traits that can be measured in a straightforward manner, opening the way to the phenotyping of molecular and mechanical aspects of shoot tropism.

Unifying model of shoot gravitropism reveals proprioception as a central feature of posture control in plants.

Gravitropism, the slow reorientation of plant growth in response to gravity, is a key determinant of the form and posture of land plants. Shoot gravitropism is triggered when statocysts sense the local angle of the growing organ relative to the gravitational field. Lateral transport of the hormone auxin to the lower side is then enhanced, resulting in differential gene expression and cell elongation causing the organ to bend. However, little is known about the dynamics, regulation, and diversity of the entire bending and straightening process. Here, we modeled the bending and straightening of a rod-like organ and compared it with the gravitropism kinematics of different organs from 11 angiosperms. We show that gravitropic straightening shares common traits across species, organs, and orders of magnitude. The minimal dynamic model accounting for these traits is not the widely cited gravisensing law but one that also takes into account the sensing of local curvature, what we describe here as a graviproprioceptive law. In our model, the entire dynamics of the bending/straightening response is described by a single dimensionless "bending number" B that reflects the ratio between graviceptive and proprioceptive sensitivities. The parameter B defines both the final shape of the organ at equilibrium and the timing of curving and straightening. B can be estimated from simple experiments, and the model can then explain most of the diversity observed in experiments. Proprioceptive sensing is thus as important as gravisensing in gravitropic control, and the B ratio can be measured as phenotype in genetic studies.

Flow-induced agitations create a granular fluid.

We fluidize a granular medium through localized stirring and probe the mechanical response of quiescent regions far away from the main flow. In these regions the material behaves like a liquid: high-density probes sink, low-density probes float at the depth given by Archimedes' law, and drag forces on moving probes scale linearly with the velocity. The fluidlike character of the material is set by agitations generated in the stirred region, suggesting a nonlocal rheology: the relation between applied stress and observed strain rate in one location depends on the strain rate in another location.

A model of collective behavior based purely on vision.

Classical models of collective behavior often take a "bird's-eye perspective," assuming that individuals have access to social information that is not directly available (e.g., the behavior of individuals outside of their field of view). Despite the explanatory success of those models, it is now thought that a better understanding needs to incorporate the perception of the individual, i.e., how internal and external information are acquired and processed. In particular, vision has appeared to be a central feature to gather external information and influence the collective organization of the group. Here, we show that a vision-based model of collective behavior is sufficient to generate organized collective behavior in the absence of spatial representation and collision. Our work suggests a different approach for the development of purely vision-based autonomous swarm robotic systems and formulates a mathematical framework for exploration of perception-based interactions and how they differ from physical ones.

Animals in Virtual Environments.

The core idea in an XR (VR/MR/AR) application is to digitally stimulate one or more sensory systems (e.g. visual, auditory, olfactory) of the human user in an interactive way to achieve an immersive experience. Since the early 2000s biologists have been using Virtual Environments (VE) to investigate the mechanisms of behavior in non-human animals including insects, fish, and mammals. VEs have become reliable tools for studying vision, cognition, and sensory-motor control in animals. In turn, the knowledge gained from studying such behaviors can be harnessed by researchers designing biologically inspired robots, smart sensors, and rnulti-agent artificial intelligence. VE for animals is becoming a widely used application of XR technology but such applications have not previously been reported in the technical literature related to XR. Biologists and computer scientists can benefit greatly from deepening interdisciplinary research in this emerging field and together we can develop new methods for conducting fundamental research in behavioral sciences and engineering. To support our argument we present this review which provides an overview of animal behavior experiments conducted in virtual environments.

Spatio-temporal integration in plant tropisms.

Tropisms, growth-driven responses to environmental stimuli, cause plant organs to respond in space and time and reorient themselves. Classical experiments from nearly a century ago reveal that plant shoots respond to the integrated history of light and gravity stimuli rather than just responding instantaneously. We introduce a temporally non-local response function for the dynamics of shoot growth formulated as an integro-differential equation whose solution allows us to qualitatively reproduce experimental observations associated with intermittent and unsteady stimuli. Furthermore, an analytic solution for the case of a pulse stimulus expresses the response function as a function of experimentally tractable variables, which we calculate for the case of the phototropic response of Arabidopsis hypocotyls. All together, our model enables us to predict tropic responses to time-varying stimuli, manifested in temporal integration phenomena, and sets the stage for the incorporation of additional effects such as multiple stimuli, gravitational sagging, etc.

Towards a framework for collective behavior in growth-driven systems, based on plant-inspired allotropic pairwise interactions.

A variety of biological systems are not motile, but sessile in nature, relying on growth as the main driver of their movement. Groups of such growing organisms can form complex structures, such as the functional architecture of growing axons, or the adaptive structure of plant root systems. These processes are not yet understood, however the decentralized growth dynamics bear similarities to the collective behavior observed in groups of motile organisms, such as flocks of birds or schools of fish. Equivalent growth mechanisms make these systems amenable to a theoretical framework inspired by tropic responses of plants, where growth is considered implicitly as the driver of the observed bending towards a stimulus. We introduce two new concepts related to plant tropisms: point tropism, the response of a plant to a nearby point signal source, and allotropism, the growth-driven response of plant organs to neighboring plants. We first analytically and numerically investigate the 2D dynamics of single organs responding to point signals fixed in space. Building on this we study pairs of organs interacting via allotropism, i.e. each organ senses signals emitted at the tip of their neighbor and responds accordingly. In the case of local sensing we find a rich state-space. We describe the different states, as well as the sharp transitions between them. We also find that the form of the state-space depends on initial conditions. This work sets the stage towards a theoretical framework for the investigation and understanding of systems of interacting growth-driven individuals.

Fluttering of growing leaves as a way to reach flatness: experimental evidence on Persea americana.

Simple leaves show unexpected growth motions: the midrib of the leaves swings periodically in association with buckling events of the leaf blade, giving the impression that the leaves are fluttering. The quantitative kinematic analysis of this motion provides information about the respective growth between the main vein and the lamina. Our three-dimensional reconstruction of an avocado tree leaf shows that the conductor of the motion is the midrib, presenting continuous oscillations and inducing buckling events on the blade. The variations in the folding angle of the leaf show that the lamina is not passive: it responds to the deformation induced by the connection to the midrib to reach a globally flat state. We model this movement as an asymmetric growth of the midrib, which directs an inhomogeneous growth of the lamina, and we suggest how the transition from the folded state to the flat state is mechanically organized.

Coupled ultradian growth and curvature oscillations during gravitropic movement in disturbed wheat coleoptiles.

To grow straight and upright, plants need to regulate actively their posture. Gravitropic movement, which occurs when plants modify their growth and curvature to orient their aerial organ against the force of gravity, is a major feature of this postural control. A recent model has shown that graviception and proprioception are sufficient to account for the gravitropic movement and subsequent organ posture demonstrated by a range of species. However, some plants, including wheat coleoptiles, exhibit a stronger regulation of posture than predicted by the model. Here, we performed an extensive kinematics study on wheat coleoptiles during a gravitropic perturbation (tilting) experiment in order to better understand this unexpectedly strong regulation. Close temporal observations of the data revealed that both perturbed and unperturbed coleoptiles showed oscillatory pulses of elongation and curvature variation that propagated from the apex to the base of their aerial organs. In perturbed coleoptiles, we discovered a non-trivial coupling between the oscillatory dynamics of curvature and elongation. The relationship between those oscillations and the postural control of the organ remains unclear, but indicates the presence of a mechanism that is capable of affecting the relationship between elongation rate, differential growth, and curvature.

Milk Fat Globules Hamper Adhesion of Enterohemorrhagic Escherichia coli to Enterocytes: In Vitro and in Vivo Evidence.

Enterohemorrhagic Escherichia coli (EHEC; E. coli) are food-borne agents associated with gastroenteritis, enterocolitis, bloody diarrhea and the hemolytic-uremic syndrome (HUS). Bovine milk glycans have been shown to contain oligosaccharides which are similar to host epithelial cell receptors and can therefore prevent bacterial adhesion. This study aimed to describe interactions between EHEC O157:H7 EDL933 and O26:H11 21765 and milk fat globules (MFGs) in raw milk and raw milk cheese, and the impact of MFGs on EHEC strains adhesion to the intestinal tract in vitro and in vivo. Both EHEC serotypes clearly associated with native bovine MFGs and significantly limited their adhesion to a co-culture of intestinal cells. The presence of MFGs in raw milk cheese had two effects on the adhesion of both EHEC serotypes to the intestinal tracts of streptomycin-treated mice. First, it delayed and reduced EHEC excretion in mouse feces for both strains. Second, the prime implantation site for both EHEC strains was 6 cm more proximal in the intestinal tracts of mice fed with contaminated cheese containing less than 5% of fat than in those fed with contaminated cheese containing 40% of fat. Feeding mice with 40% fat cheese reduced the intestinal surface contaminated with EHEC and may therefore decrease severity of illness.

Stochastic processes in gravitropism.

In this short review we focus on the role of noise in gravitropism of plants - the reorientation of plants according to the direction of gravity. We briefly introduce the conventional picture of static gravisensing in cells specialized in sensing. This model hinges on the sedimentation of statoliths (high in density and mass relative to other organelles) to the lowest part of the sensing cell. We then present experimental observations that cannot currently be understood within this framework. Lastly we introduce some current alternative models and directions that attempt to incorporate and interpret these experimental observations, including: (i) dynamic sensing, where gravisensing is suggested to be enhanced by stochastic events due to thermal and mechanical noise. These events both effectively lower the threshold of response, and lead to small-distance sedimentation, allowing amplification, and integration of the signal. (ii) The role of the cytoskeleton in signal-to-noise modulation and (iii) in signal transduction. In closing, we discuss directions that seem to either not have been explored, or that are still poorly understood.

A unifying modeling of plant shoot gravitropism with an explicit account of the effects of growth.

Gravitropism, the slow reorientation of plant growth in response to gravity, is a major determinant of the form and posture of land plants. Recently a universal model of shoot gravitropism, the AC model, was presented, in which the dynamics of the tropic movement is only determined by the conflicting controls of (1) graviception that tends to curve the plants toward the vertical, and (2) proprioception that tends to keep the stem straight. This model was found to be valid for many species and over two orders of magnitude of organ size. However, the motor of the movement, the elongation, was purposely neglected in the AC model. If growth effects are to be taken into account, it is necessary to consider the material derivative, i.e., the rate of change of curvature bound to expanding and convected organ elements. Here we show that it is possible to rewrite the material equation of curvature in a compact simplified form that directly expresses the curvature variation as a function of the median elongation and of the distribution of the differential growth. By using this extended model, called the ACE model, growth is found to have two main destabilizing effects on the tropic movement: (1) passive orientation drift, which occurs when a curved element elongates without differential growth, and (2) fixed curvature, when an element leaves the elongation zone and is no longer able to actively change its curvature. By comparing the AC and ACE models to experiments, these two effects are found to be negligible. Our results show that the simplified AC mode can be used to analyze gravitropism and posture control in actively elongating plant organs without significant information loss.