Stereo Camera Setup for 360° Digital Image Correlation to Reveal Smart Structures of Hakea Fruits.

About forty years after its first application, digital image correlation (DIC) has become an established method for measuring surface displacements and deformations of objects under stress. To date, DIC has been used in a variety of in vitro and in vivo studies to biomechanically characterise biological samples in order to reveal biomimetic principles. However, when surfaces of samples strongly deform or twist, they cannot be thoroughly traced. To overcome this challenge, different DIC setups have been developed to provide additional sensor perspectives and, thus, capture larger parts of an object's surface. Herein, we discuss current solutions for this multi-perspective DIC, and we present our own approach to a 360° DIC system based on a single stereo-camera setup. Using this setup, we are able to characterise the desiccation-driven opening mechanism of two woody Hakea fruits over their entire surfaces. Both the breaking mechanism and the actuation of the two valves in predominantly dead plant material are models for smart materials. Based on these results, an evaluation of the setup for 360° DIC regarding its use in deducing biomimetic principles is given. Furthermore, we propose a way to improve and apply the method for future measurements.

Elastic pinch biomechanisms can yield consistent launch speeds regardless of projectile mass.

Energetic trade-offs are particularly pertinent to bio-ballistic systems which impart energy to projectiles exclusively during launch. We investigated such trade-offs in the spring-propelled seeds of Loropetalum chinense, Hamamelis virginiana and Fortunearia sinensis. Using similar seed-shooting mechanisms, fruits of these confamilial plants (Hamamelidaceae) span an order of magnitude in spring and seed mass. We expected that as seed mass increases, launch speed decreases. Instead, launch speed was relatively constant regardless of seed mass. We tested if fruits shoot larger seeds by storing more elastic potential energy (PE). Spring mass and PE increased as seed mass increased (in order of increasing seed mass: L. chinense, H. virginiana, F. sinensis). As seed mass to spring mass ratio increased (ratios: H. virginiana = 0.50, F. sinensis = 0.65, L. chinense = 0.84), mass-specific PE storage increased. The conversion efficiency of PE to seed kinetic energy (KE) decreased with increasing fruit mass. Therefore, similar launch speeds across scales occurred because (i) larger fruits stored more PE and (ii) smaller fruits had higher mass-specific PE storage and improved PE to KE conversion. By examining integrated spring and projectile mechanics in our focal species, we revealed diverse, energetic scaling strategies relevant to spring-propelled systems navigating energetic trade-offs.

Force Generation in the Coiling Tendrils of Passiflora caerulea.

Tendrils of climbing plants coil along their length, thus forming a striking helical spring and generating tensional forces. It is found that, for tendrils of the passion flower Passiflora caerulea, the generated force lies in the range of 6-140 mN, which is sufficient to lash the plant tightly to its substrate. Further, it is revealed that the generated force strongly correlates with the water status of the plant. Based on a combination of in situ force measurements with anatomical investigations and dehydration-rehydration experiments on both entire tendril segments and isolated lignified tissues, a two-phasic mechanism for spring formation is proposed. First, during the free coiling phase, the center of the tendril begins to lignify unilaterally. At this stage, both the generated tension and the stability of the form of the spring still depend on turgor pressure. The unilateral contraction of a bilayer as being the possible driving force for the tendril coiling motion is discussed. Second, in a stabilization phase, the entire center of the coiled tendril lignifies, stiffening the spring and securing its function irrespective of its hydration status.

How dehydration affects stem bending stiffness and leaf toughness after sampling of the liana Amphilophium crucigerum (L.) L.G.Lohmann (Bignoniaceae) / Como a desidratação afeta a flexibilidade do caule e a tenacidade das folhas pós-coleta na liana Amphilophium crucigerum (L.) L.G.Lohmann (Bignoniaceae)

Lianas are woody climbers and their stems and leaves deal with different environmental pressures such as resistance to mechanical damage and dehydration. The damage resistance of plants can be biomechanically evaluated by their stiffness, bending and toughness. Despite the well-known relationship between physical resistance and moisture of plant organs in woody plants, this relationship is uncertain and has not been previously evaluated in lianas. Thus, this study investigated experimentally the effect of stems and leaf dehydration on the structural Young's modulus in the stem and fracture toughness in leaves across time in the liana Amphilophium crucigerum (Bignoniaceae). Ten stem and leaf samples were collected and assigned to two distinct conditions: (i) samples kept moist and (ii) samples underwent gradual dehydration with natural moisture loss by air exposition. Successive measures of structural Young's modulus and fracture toughness were taken every 4 hours during a 48-hour period for both conditions. Stem and leaf samples which underwent gradual dehydration showed greater bending stiffness and fracture toughness, respectively, while the samples kept moist presented no changes in any studied biomechanical features during the entire experiment. We concluded that the moisture of both stem and leaf samples are critical factors to estimate the biomechanical properties of lianas stem and leaves.

Lianas são trepadeiras lenhosas e seus caules e folhas lidam com diferentes pressões ambientais, como a resistência aos danos mecânicos e de desidratação. A resistência ao dano das plantas pode ser biomecanicamente avaliada pelas propiedades de dureza, flexão e tenacidade. Apesar da conhecida relação entre resistência física e umidade dos órgãos das plantas em espécies lenhosas, essa relação não foi avaliada anteriormente e é incerta em lianas. Assim, este estudo investigou experimentalmente o efeito da desidratação de caules e folhas na estimativa do módulo estrutural de Young do caule e da tenacidade à fratura da folha ao longo do tempo, na liana Amphilophium crucigerum (Bignoniaceae). Dez amostras de caules e folhas foram coletadas e distribuídas em duas condições distintas: (i) amostras mantidas úmidas e (ii) amostras em processo de desidratação gradativa com perda natural de umidade quando expostas ao ar. Medidas sucessivas do módulo de Young e da resistência à fratura dos órgãos foram tomadas a cada 4 horas durante um período de 48 horas em ambas as condições. Amostras de caule e folhas que sofreram desidratação gradual apresentaram maior rigidez à flexão e tenacidade à fratura, respectivamente, enquanto as amostras mantidas úmidas não alteraram essas características durante o experimento. Concluímos que a umidade das amostras de caules e folhas em lianas também é um fator crítico para estimar as propriedades biomecânicas desses órgãos em seu ambiente natural.

Morphological Computation in Plant Seeds for a New Generation of Self-Burial and Flying Soft Robots.

Plants have evolved different mechanisms to disperse from parent plants and improve germination to sustain their survival. The study of seed dispersal mechanisms, with the related structural and functional characteristics, is an active research topic for ecology, plant diversity, climate change, as well as for its relevance for material science and engineering. The natural mechanisms of seed dispersal show a rich source of robust, highly adaptive, mass and energy efficient mechanisms for optimized passive flying, landing, crawling and drilling. The secret of seeds mobility is embodied in the structural features and anatomical characteristics of their tissues, which are designed to be selectively responsive to changes in the environmental conditions, and which make seeds one of the most fascinating examples of morphological computation in Nature. Particularly clever for their spatial mobility performance, are those seeds that use their morphology and structural characteristics to be carried by the wind and dispersed over great distances (i.e. "winged" and "parachute" seeds), and seeds able to move and penetrate in soil with a self-burial mechanism driven by their hygromorphic properties and morphological features. By looking at their motion mechanisms, new design principles can be extracted and used as inspiration for smart artificial systems endowed with embodied intelligence. This mini-review systematically collects, for the first time together, the morphological, structural, biomechanical and aerodynamic information from selected plant seeds relevant to take inspiration for engineering design of soft robots, and discusses potential future developments in the field across material science, plant biology, robotics and embodied intelligence.

Structure and Biomechanics during Xylem Vessel Transdifferentiation in Arabidopsis thaliana.

Individual plant cells are the building blocks for all plantae and artificially constructed plant biomaterials, like biocomposites. Secondary cell walls (SCWs) are a key component for mediating mechanical strength and stiffness in both living vascular plants and biocomposite materials. In this paper, we study the structure and biomechanics of cultured plant cells during the cellular developmental stages associated with SCW formation. We use a model culture system that induces transdifferentiation of Arabidopsis thaliana cells to xylem vessel elements, upon treatment with dexamethasone (DEX). We group the transdifferentiation process into three distinct stages, based on morphological observations of the cell walls. The first stage includes cells with only a primary cell wall (PCW), the second covers cells that have formed a SCW, and the third stage includes cells with a ruptured tonoplast and partially or fully degraded PCW. We adopt a multi-scale approach to study the mechanical properties of cells in these three stages. We perform large-scale indentations with a micro-compression system in three different osmotic conditions. Atomic force microscopy (AFM) nanoscale indentations in water allow us to isolate the cell wall response. We propose a spring-based model to deconvolve the competing stiffness contributions from turgor pressure, PCW, SCW and cytoplasm in the stiffness of differentiating cells. Prior to triggering differentiation, cells in hypotonic pressure conditions are significantly stiffer than cells in isotonic or hypertonic conditions, highlighting the dominant role of turgor pressure. Plasmolyzed cells with a SCW reach similar levels of stiffness as cells with maximum turgor pressure. The stiffness of the PCW in all of these conditions is lower than the stiffness of the fully-formed SCW. Our results provide the first experimental characterization of the mechanics of SCW formation at single cell level.

Assembly of a simple scalable device for micromechanical testing of plant tissues.

Tensile testing is widely used to evaluate the mechanical properties of biological materials including soft primary plant tissues. Commercially available platforms for tensile testing are often expensive and limited in customizability. In this chapter, we provide a guide for the assembly and use of a simple and low-cost micromechanical testing apparatus suitable for research and educational purposes. The build of the setup is presented with scalability and universality in mind and is based on a do-it-yourself mind frame towards mechanical tests on plant organs and tissues. We discuss hardware and software requirements with practical details on required components, device calibration and a script to run the device. Further, we provide an example in which the device was used for the uniaxial tensile test of onion epidermis.

Snapping mechanics of the Venus flytrap (Dionaea muscipula).

The mechanical principles for fast snapping in the iconic Venus flytrap are not yet fully understood. In this study, we obtained time-resolved strain distributions via three-dimensional digital image correlation (DIC) for the outer and inner trap-lobe surfaces throughout the closing motion. In combination with finite element models, the various possible contributions of the trap tissue layers were investigated with respect to the trap's movement behavior and the amount of strain required for snapping. Supported by in vivo experiments, we show that full trap turgescence is a mechanical-physiological prerequisite for successful (fast and geometrically correct) snapping, driven by differential tissue changes (swelling, shrinking, or no contribution). These are probably the result of the previous accumulation of internal hydrostatic pressure (prestress), which is released after trap triggering. Our research leads to an in-depth mechanical understanding of a complex plant movement incorporating various actuation principles.

Bladderworts, the smallest known suction feeders, generate inertia-dominated flows to capture prey.

Aquatic bladderworts (Utricularia gibba and U. australis) capture zooplankton in mechanically triggered underwater traps. With characteristic dimensions less than 1 mm, the trapping structures are among the smallest known to capture prey by suction, a mechanism that is not effective in the creeping-flow regime where viscous forces prevent the generation of fast and energy-efficient suction flows. To understand what makes suction feeding possible on the small scale of bladderwort traps, we characterised their suction flows experimentally (using particle image velocimetry) and mathematically (using computational fluid dynamics and analytical mathematical models). We show that bladderwort traps avoid the adverse effects of creeping flow by generating strong, fast-onset suction pressures. Our findings suggest that traps use three morphological adaptations: the trap walls' fast release of elastic energy ensures strong and constant suction pressure; the trap door's fast opening ensures effectively instantaneous onset of suction; the short channel leading into the trap ensures undeveloped flow, which maintains a wide effective channel diameter. Bladderwort traps generate much stronger suction flows than larval fish with similar gape sizes because of the traps' considerably stronger suction pressures. However, bladderworts' ability to generate strong suction flows comes at considerable energetic expense.

Kinematics Governing Mechanotransduction in the Sensory Hair of the Venus flytrap.

Insects fall prey to the Venus flytrap (Dionaea muscipula) when they touch the sensory hairs located on the flytrap lobes, causing sudden trap closure. The mechanical stimulus imparted by the touch produces an electrical response in the sensory cells of the trigger hair. These cells are found in a constriction near the hair base, where a notch appears around the hair's periphery. There are mechanosensitive ion channels (MSCs) in the sensory cells that open due to a change in membrane tension; however, the kinematics behind this process is unclear. In this study, we investigate how the stimulus acts on the sensory cells by building a multi-scale hair model, using morphometric data obtained from µ-CT scans. We simulated a single-touch stimulus and evaluated the resulting cell wall stretch. Interestingly, the model showed that high stretch values are diverted away from the notch periphery and, instead, localized in the interior regions of the cell wall. We repeated our simulations for different cell shape variants to elucidate how the morphology influences the location of these high-stretch regions. Our results suggest that there is likely a higher mechanotransduction activity in these 'hotspots', which may provide new insights into the arrangement and functioning of MSCs in the flytrap.

A mathematical model explores the contributions of bending and stretching forces to shoot gravitropism in Arabidopsis.

Plant shoot gravitropism is a complex phenomenon resulting from gravity sensing, curvature sensing (proprioception), the ability to uphold self-weight and growth. Although recent data analysis and modelling have revealed the detailed morphology of shoot bending, the relative contribution of bending force (derived from the gravi-proprioceptive response) and stretching force (derived from shoot axial growth) behind gravitropism remains poorly understood. To address this gap, we combined morphological data with a theoretical model to analyze shoot bending in wild-type and lazy1-like 1 mutant Arabidopsis thaliana. Using data from actual bending events, we searched for model parameters that minimized discrepancies between the data and mathematical model. The resulting model suggests that both the bending force and the stretching force differ significantly between the wild type and mutant. We discuss the implications of the mechanical forces associated with differential cell growth and present a plausible mechanical explanation of shoot gravitropism.

Root shape adaptation to mechanical stress derived from unidirectional vibrations in Populus nigra.

While it is known that plant roots can change their shapes to the stress direction, it remains unclear if the root orientation can change as a means for mechanical reinforcement. When stress in form of a unidirectional vibration is applied to cuttings of Populus nigra for 5 min a day over a period of 20 days, the root system architecture changes. The contribution of roots with a diameter larger than 0.04 cm increases, while the allocation to roots smaller than 0.03 cm decreases. In addition to the root diameter allocation, the root orientation in the stem proximity was analyzed by appearance and with a nematic tensor analysis in an attempt to calculate the average root orientation. The significant different allocation to roots with a larger diameter, and the tendency of roots to align in the vicinity of the stress axis (not significantly different), are indicating a mechanical reinforcement to cope with the received strain. This work indicates an adaptive root system architecture and a possible adaptive root orientation for mechanical reinforcement.

The Structure of the Barley Husk Influences Its Resistance to Mechanical Stress.

This paper explores the links between genotype, plant development, plant structure and plant material properties. The barley husk has two organs, the lemma and the palea, which protect the grain. When the husk is exposed to mechanical stress, such as during harvesting, it can be damaged or detached. This is known as grain skinning, which is detrimental to grain quality and has a significant economic impact on industry. This study focused on the lemma, the husk organ which is most susceptible to grain skinning. This study tested three hypotheses: (1) genotype and plant development determine lemma structure, (2) lemma structure influences the material properties of the lemma, and (3) the material properties of the lemma determine grain skinning risk. The effect of genotype was investigated by using plant material from four malting barley varieties: two with a high risk of grain skinning, two with a low risk. Plant material was assessed at two stages of plant development (anthesis, GS 65; grain filling, GS 77). Structure was assessed using light microscopy to measure three physiological features: thickness, vasculature and cell area. Material properties were approximated using a controlled impact assay and by analyzing fragmentation behavior. Genotype had a significant effect on lemma structure and material properties from anthesis. This indicates that differences between genotypes were established during floral development. The lemma was significantly thinner in high risk genotypes, compared to low risk genotypes. Consequently, in high risk genotypes, the lemma was significantly more likely to fragment. This indicates a relationship between reduced lemma thickness and increased fragmentation. Traditionally, a thin husk has been considered beneficial for malting quality, due to an association with malt extract. However, this study finds a thin lemma is less resistant to mechanical stress. This may explain the differences in grain skinning risk in the genotypes studied.

A seed flying like a bullet: ballistic seed dispersal in Chinese witch-hazel (Hamamelis mollis OLIV., Hamamelidaceae).

The fruits of Chinese witch-hazel (Hamamelis mollis, Hamamelidaceae) act as 'drying squeeze catapults', shooting their seeds several metres away. During desiccation, the exocarp shrinks and splits open, and subsequent endocarp deformation is a complex three-dimensional shape change, including formation of dehiscence lines, opening of the apical part and formation of a constriction at the middle part. Owing to the constriction forming, mechanical pressure is increasingly applied on the seed until ejection. We describe a structural latch system consisting of connective cellular structures between endocarp and seed, which break with a distinct cracking sound upon ejection. A maximum seed velocity of 12.3 m s-1, maximum launch acceleration of 19 853 m s-2 (approx. 2000g) and maximum seed rotational velocity of 25 714 min-1 were measured. We argue that miniscule morphological differences between the inner endocarp surface and seed, which features a notable ridge, are responsible for putting spin on the seed. This hypothesis is further corroborated by the observation that there is no preferential seed rotation direction among fruits. Our findings show that H. mollis has evolved similar mechanisms for stabilizing a 'shot out' seed as humans use for stabilizing rifle bullets and are discussed in an ecological (dispersal biology), biomechanical (seed ballistics) and functional-morphological (fine-tuning and morphospace of functional endocarps) contexts, and promising additional aspects for future studies are proposed.

Comparative analysis of pollen release biomechanics in Thalictrum: implications for evolutionary transitions between animal and wind pollination.

Transitions from animal to wind pollination have occurred repeatedly in flowering plants, driven by structural and biomechanical modifications to flowers. But the initial changes promoting wind pollination are poorly understood, especially those required to release pollen into airflows - the critical first stage of wind pollination. Using a wind tunnel, we performed a comparative study of pollen release biomechanics in 36 species of animal- and wind-pollinated Thalictrum. We quantified pollination syndromes and stamen natural frequency (fn ), a key vibration parameter, to determine if floral traits reliably predicted pollen release probability. We then investigated if pollen release was caused by wind-induced resonance vibration of stamens. We detected wind-induced stamen resonance in 91% of species and a strong effect of stamen acceleration on pollen release, inversely driven by fn . However, unlike fn , pollination syndromes did not reliably predict the probability of pollen release among species. Our results directly link fn to the capacity of stamens to release pollen by wind and suggest that structural mechanisms reducing fn are likely to be important for initiating transitions from animal to wind pollination. Our inability to predict the probability of pollen release based on pollination syndromes suggests diverse phenotypic trajectories from animal to wind pollination.

How water flow, geometry, and material properties drive plant movements.

Plants are dynamic. They adjust their shape for feeding, defence, and reproduction. Such plant movements are critical for their survival. We present selected examples covering a range of movements from single cell to tissue level and over a range of time scales. We focus on reversible turgor-driven shape changes. Recent insights into the mechanisms of stomata, bladderwort, the waterwheel, and the Venus flytrap are presented. The underlying physical principles (turgor, osmosis, membrane permeability, wall stress, snap buckling, and elastic instability) are highlighted, and advances in our understanding of these processes are summarized.

Kinematical, Structural and Mechanical Adaptations to Desiccation in Poikilohydric Ramonda myconi (Gesneriaceae).

Resurrection plants have fascinated scientists since centuries as they can fully recover from cellular water contents below 10%, concomitantly showing remarkable leaf folding motions. While physiological adaptations have been meticulously investigated, the understanding of structural and mechanical adaptations of this phenomenon is scarce. Using imaging and bending techniques during dehydration-rehydration experiments, morphological, anatomical, and biomechanical properties of desiccation-tolerant Ramonda myconi are examined, and selected structural adaptations are compared to those of homoiohydrous Monophyllaea horsfieldii (both Gesneriaceae). At low water availability, intact and cut-off R. myconi leaves undergo considerable morphological alterations, which are fully and repeatedly reversible upon rehydration. Furthermore, their petioles show a triphasic mechanical behavior having a turgor-based structural stability at high (Phase 1), a flexible mechanically state at intermediate (Phase 2) and a material-based stability at low water contents (Phase 3). Lastly, manipulation experiments with cut-off plant parts revealed that both the shape alterations of individual structures, as well as, the general leaf kinematics largely rely on passive swelling and shrinking processes. Taken together, R. myconi possesses structural and mechanical adaptations to desiccation (in addition to physiological adaptations), which may mainly be passively driven by its water status influenced by the water fluctuations in its surroundings.

Resistance from agar medium impacts the helical growth of Arabidopsis primary roots.

Agar is widely used in studies of root growth since it can be mixed at different concentrations to impact mechanical impedance. At high concentrations (1.2-1.5%), growth of Arabidopsis roots has been found to be wavy, but little research has explored this behavior based on a quantitative understanding of mechanical behavior. To this end, agar media with concentration ranging from 0.5% to 1.2% were prepared to produce gradient resistance during root penetration, and Young's moduli and penetrometer resistance were tested. Arabidopsis roots were then cultivated in these agar media with gradient stiffness. The result showed that Young's modulus increased linearly with the increase of concentration of agar media. For Arabidopsis primary roots, it was preferred to develop a helical pattern in agar media with concentration from 0.5% to 1.0%. As stiffness of agar increased, the percentage of helical roots and helix diameters in each agar medium declined; root lengths and auxin distributions showed variety. We demonstrate that the size of helical deformation decreases with agar stiffness as expected by theoretical analysis based on a combination of growth-induced mechanical buckling. In conclusion, the resistance from agar media impacts the properties of root helix, and helical roots growth is driven by growth force. Growth force and external mechanical forces contribute to root phenotypes in Arabidopsis.

Gravisensors in plant cells behave like an active granular liquid.

Plants are able to sense and respond to minute tilt from the vertical direction of the gravity, which is key to maintain their upright posture during development. However, gravisensing in plants relies on a peculiar sensor made of microsize starch-filled grains (statoliths) that sediment and form tiny granular piles at the bottom of the cell. How such a sensor can detect inclination is unclear, as granular materials like sand are known to display flow threshold and finite avalanche angle due to friction and interparticle jamming. Here, we address this issue by combining direct visualization of statolith avalanches in plant cells and experiments in biomimetic cells made of microfluidic cavities filled with a suspension of heavy Brownian particles. We show that, despite their granular nature, statoliths move and respond to the weakest angle, as a liquid clinometer would do. Comparison between the biological and biomimetic systems reveals that this liquid-like behavior comes from the cell activity, which agitates statoliths with an apparent temperature one order of magnitude larger than actual temperature. Our results shed light on the key role of active fluctuations of statoliths for explaining the remarkable sensitivity of plants to inclination. Our study also provides support to a recent scenario of gravity perception in plants, by bridging the active granular rheology of statoliths at the microscopic level to the macroscopic gravitropic response of the plant.