Completing the picture of field-grown cereal crops: a new method for detailed leaf surface models in wheat.

BACKGROUND: The leaf angle distribution (LAD) is an important structural parameter of agricultural crops that influences light interception, radiation fluxes and consequently plant performance. Therefore, LAD and its parametrized form, the Beta distribution, is used in many photosynthesis models. However, in field cultivations, these parameters are difficult to assess and cereal crops in particular pose challenges since their leaves are thin, flexible, and often bent and twisted around their own axis. To our knowledge, there is only a very limited set of methods currently available to calculate LADs of field-grown cereal crops that explicitly takes these special morphological properties into account. RESULTS: In this study, a new processing pipeline is introduced that allows for the generation of realistic leaf surface models and the analysis of LADs of field-grown cereal crops from 3D point clouds. The data acquisition is based on a convenient stereo imaging setup. The approach was validated with different artificial targets and results on the accuracy of the 3D reconstruction, leaf surface modeling and calculated LAD are given. The mean error of the 3D reconstruction was below 1 mm for an inclination angle range between 0° and 75° and the leaf surface could be quantified with an average accuracy of 90%. The concordance correlation coefficient (CCC) of 99.6% (p-value = [Formula: see text]) indicated a high correlation between the reconstructed inclination angle and the identity line. The LADs for bent leaves were reconstructed with a mean error of 0.21° and a standard deviation of 1.55°. As an additional parameter, the insertion angle was reconstructed for the artificial leaf model with an average error < 5°. Finally, the method was tested with images of field-grown cereal crops and Beta functions were approximated from the calculated LADs. The mean CCC between reconstructed LAD and calculated Beta function was 0.66. According to Cohen, this indicates a high correlation. CONCLUSION: This study shows that our image processing pipeline can reconstruct the complex leaf shape of cereal crops from stereo images. The high accuracy of the approach was demonstrated with several validation experiments including artificial leaf targets. The derived leaf models were used to calculate LADs for artificial leaves and naturally grown cereal crops. This helps to better understand the influence of the canopy structure on light absorption and plant performance and allows for a more precise parametrization of photosynthesis models via the derived Beta distributions.

Plant Growth Promotion and Heat Stress Amelioration in Arabidopsis Inoculated with Paraburkholderia phytofirmans PsJN Rhizobacteria Quantified with the GrowScreen-Agar II Phenotyping Platform.

High temperatures inhibit plant growth. A proposed strategy for improving plant productivity under elevated temperatures is the use of plant growth-promoting rhizobacteria (PGPR). While the effects of PGPR on plant shoots have been extensively explored, roots-particularly their spatial and temporal dynamics-have been hard to study, due to their below-ground nature. Here, we characterized the time- and tissue-specific morphological changes in bacterized plants using a novel non-invasive high-resolution plant phenotyping and imaging platform-GrowScreen-Agar II. The platform uses custom-made agar plates, which allow air exchange to occur with the agar medium and enable the shoot to grow outside the compartment. The platform provides light protection to the roots, the exposure of it to the shoots, and the non-invasive phenotyping of both organs. Arabidopsis thaliana, co-cultivated with Paraburkholderia phytofirmans PsJN at elevated and ambient temperatures, showed increased lengths, growth rates, and numbers of roots. However, the magnitude and direction of the growth promotion varied depending on root type, timing, and temperature. The root length and distribution per depth and according to time was also influenced by bacterization and the temperature. The shoot biomass increased at the later stages under ambient temperature in the bacterized plants. The study offers insights into the timing of the tissue-specific, PsJN-induced morphological changes and should facilitate future molecular and biochemical studies on plant-microbe-environment interactions.

N-dependent dynamics of root growth and nitrate and ammonium uptake are altered by the bacterium Herbaspirillum seropedicae in the cereal model Brachypodium distachyon.

Nitrogen (N) fixation in cereals by root-associated bacteria is a promising solution for reducing use of chemical N fertilizers in agriculture. However, plant and bacterial responses are unpredictable across environments. We hypothesized that cereal responses to N-fixing bacteria are dynamic, depending on N supply and time. To quantify the dynamics, a gnotobiotic, fabricated ecosystem (EcoFAB) was adapted to analyse N mass balance, to image shoot and root growth, and to measure gene expression of Brachypodium distachyon inoculated with the N-fixing bacterium Herbaspirillum seropedicae. Phenotyping throughput of EcoFAB-N was 25-30 plants h-1 with open software and imaging systems. Herbaspirillum seropedicae inoculation of B. distachyon shifted root and shoot growth, nitrate versus ammonium uptake, and gene expression with time; directions and magnitude depended on N availability. Primary roots were longer and root hairs shorter regardless of N, with stronger changes at low N. At higher N, H. seropedicae provided 11% of the total plant N that came from sources other than the seed or the nutrient solution. The time-resolved phenotypic and molecular data point to distinct modes of action: at 5 mM NH4NO3 the benefit appears through N fixation, while at 0.5 mM NH4NO3 the mechanism appears to be plant physiological, with H. seropedicae promoting uptake of N from the root medium.Future work could fine-tune plant and root-associated microorganisms to growth and nutrient dynamics.

Assessing the Storage Root Development of Cassava with a New Analysis Tool.

Storage roots of cassava plants crops are one of the main providers of starch in many South American, African, and Asian countries. Finding varieties with high yields is crucial for growing and breeding. This requires a better understanding of the dynamics of storage root formation, which is usually done by repeated manual evaluation of root types, diameters, and their distribution in excavated roots. We introduce a newly developed method that is capable to analyze the distribution of root diameters automatically, even if root systems display strong variations in root widths and clustering in high numbers. An application study was conducted with cassava roots imaged in a video acquisition box. The root diameter distribution was quantified automatically using an iterative ridge detection approach, which can cope with a wide span of root diameters and clustering. The approach was validated with virtual root models of known geometries and then tested with a time-series of excavated root systems. Based on the retrieved diameter classes, we show plausibly that the dynamics of root type formation can be monitored qualitatively and quantitatively. We conclude that this new method reliably determines important phenotypic traits from storage root crop images. The method is fast and robustly analyses complex root systems and thereby applicable in high-throughput phenotyping and future breeding.

Plant Screen Mobile: an open-source mobile device app for plant trait analysis.

BACKGROUND: The development of leaf area is one of the fundamental variables to quantify plant growth and physiological function and is therefore widely used to characterize genotypes and their interaction with the environment. To date, analysis of leaf area often requires elaborate and destructive measurements or imaging-based methods accompanied by automation that may result in costly solutions. Consequently in recent years there is an increasing trend towards simple and affordable sensor solutions and methodologies. A major focus is currently on harnessing the potential of applications developed for smartphones that provide access to analysis tools to a wide user basis. However, most existing applications entail significant manual effort during data acquisition and analysis. RESULTS: With the development of Plant Screen Mobile we provide a suitable smartphone solution for estimating digital proxies of leaf area and biomass in various imaging scenarios in the lab, greenhouse and in the field. To distinguish between plant tissue and background the core of the application comprises different classification approaches that can be parametrized by users delivering results on-the-fly. We demonstrate the practical applications of computing projected leaf area based on two case studies with Eragrostis and Musa plants. These studies showed highly significant correlations with destructive measurements of leaf area and biomass from both ground truth measurements and estimations from well-established screening systems. CONCLUSIONS: We show that a smartphone together with our analysis tool Plant Screen Mobile is a suitable platform for rapid quantification of leaf and shoot development of various plant architectures. Beyond the estimation of projected leaf area the app can also be used to quantify color and shape parameters of other plant material including seeds and flowers.

Quantitative Estimation of Leaf Heat Transfer Coefficients by Active Thermography at Varying Boundary Layer Conditions.

Quantifying heat and mass exchanges processes of plant leaves is crucial for detailed understanding of dynamic plant-environment interactions. The two main components of these processes, convective heat transfer, and transpiration, are inevitably coupled as both processes are restricted by the leaf boundary layer. To measure leaf heat capacity and leaf heat transfer coefficient, we thoroughly tested and applied an active thermography method that uses a transient heat pulse to compute τ, the time constant of leaf cooling after release of the pulse. We validated our approach in the laboratory on intact leaves of spring barley (Hordeum vulgare) and common bean (Phaseolus vulgaris), and measured τ-changes at different boundary layer conditions.By modeling the leaf heat transfer coefficient with dimensionless numbers, we could demonstrate that τ improves our ability to close the energy budget of plant leaves and that modeling of transpiration requires considerations of convection. Applying our approach to thermal images we obtained spatio-temporal maps of τ, providing observations of local differences in thermal responsiveness of leaf surfaces. We propose that active thermography is an informative methodology to measure leaf heat transfer and derive spatial maps of thermal responsiveness of leaves contributing to improve models of leaf heat transfer processes.

Fast High Resolution Volume Carving for 3D Plant Shoot Reconstruction.

Volume carving is a well established method for visual hull reconstruction and has been successfully applied in plant phenotyping, especially for 3d reconstruction of small plants and seeds. When imaging larger plants at still relatively high spatial resolution (≤1 mm), well known implementations become slow or have prohibitively large memory needs. Here we present and evaluate a computationally efficient algorithm for volume carving, allowing e.g., 3D reconstruction of plant shoots. It combines a well-known multi-grid representation called "Octree" with an efficient image region integration scheme called "Integral image." Speedup with respect to less efficient octree implementations is about 2 orders of magnitude, due to the introduced refinement strategy "Mark and refine." Speedup is about a factor 1.6 compared to a highly optimized GPU implementation using equidistant voxel grids, even without using any parallelization. We demonstrate the application of this method for trait derivation of banana and maize plants.

Sun-induced chlorophyll fluorescence from high-resolution imaging spectroscopy data to quantify spatio-temporal patterns of photosynthetic function in crop canopies.

Passive detection of sun-induced chlorophyll fluorescence (SIF) using spectroscopy has been proposed as a proxy to quantify changes in photochemical efficiency at canopy level under natural light conditions. In this study, we explored the use of imaging spectroscopy to quantify spatio-temporal dynamics of SIF within crop canopies and its sensitivity to track patterns of photosynthetic activity originating from the interaction between vegetation structure and incoming radiation as well as variations in plant function. SIF was retrieved using the Fraunhofer Line Depth (FLD) principle from imaging spectroscopy data acquired at different time scales a few metres above several crop canopies growing under natural illumination. We report the first maps of canopy SIF in high spatial resolution. Changes of SIF were monitored at different time scales ranging from quick variations under induced stress conditions to seasonal dynamics. Natural changes were primarily determined by varying levels and distribution of photosynthetic active radiation (PAR). However, this relationship changed throughout the day demonstrating an additional physiological component modulating spatio-temporal patterns of SIF emission. We successfully used detailed SIF maps to track changes in the canopy's photochemical activity under field conditions, providing a new tool to evaluate complex patterns of photosynthesis within the canopy.

The leaf angle distribution of natural plant populations: assessing the canopy with a novel software tool.

BACKGROUND: Three-dimensional canopies form complex architectures with temporally and spatially changing leaf orientations. Variations in canopy structure are linked to canopy function and they occur within the scope of genetic variability as well as a reaction to environmental factors like light, water and nutrient supply, and stress. An important key measure to characterize these structural properties is the leaf angle distribution, which in turn requires knowledge on the 3-dimensional single leaf surface. Despite a large number of 3-d sensors and methods only a few systems are applicable for fast and routine measurements in plants and natural canopies. A suitable approach is stereo imaging, which combines depth and color information that allows for easy segmentation of green leaf material and the extraction of plant traits, such as leaf angle distribution. RESULTS: We developed a software package, which provides tools for the quantification of leaf surface properties within natural canopies via 3-d reconstruction from stereo images. Our approach includes a semi-automatic selection process of single leaves and different modes of surface characterization via polygon smoothing or surface model fitting. Based on the resulting surface meshes leaf angle statistics are computed on the whole-leaf level or from local derivations. We include a case study to demonstrate the functionality of our software. 48 images of small sugar beet populations (4 varieties) have been analyzed on the base of their leaf angle distribution in order to investigate seasonal, genotypic and fertilization effects on leaf angle distributions. We could show that leaf angle distributions change during the course of the season with all varieties having a comparable development. Additionally, different varieties had different leaf angle orientation that could be separated in principle component analysis. In contrast nitrogen treatment had no effect on leaf angles. CONCLUSIONS: We show that a stereo imaging setup together with the appropriate image processing tools is capable of retrieving the geometric leaf surface properties of plants and canopies. Our software package provides whole-leaf statistics but also a local estimation of leaf angles, which may have great potential to better understand and quantify structural canopy traits for guided breeding and optimized crop management.

Organization of excitable dynamics in hierarchical biological networks.

This study investigates the contributions of network topology features to the dynamic behavior of hierarchically organized excitable networks. Representatives of different types of hierarchical networks as well as two biological neural networks are explored with a three-state model of node activation for systematically varying levels of random background network stimulation. The results demonstrate that two principal topological aspects of hierarchical networks, node centrality and network modularity, correlate with the network activity patterns at different levels of spontaneous network activation. The approach also shows that the dynamic behavior of the cerebral cortical systems network in the cat is dominated by the network's modular organization, while the activation behavior of the cellular neuronal network of Caenorhabditis elegans is strongly influenced by hub nodes. These findings indicate the interaction of multiple topological features and dynamic states in the function of complex biological networks.

Consistency analysis of metabolic correlation networks.

BACKGROUND: Metabolic correlation networks are derived from the covariance of metabolites in replicates of metabolomics experiments. They constitute an interesting intermediate between topology (i.e. the system's architecture defined by the set of reactions between metabolites) and dynamics (i.e. the metabolic concentrations observed as fluctuations around steady-state values in the metabolic network). RESULTS: Here we analyze, how such a correlation network changes over time, and compare the relative positions of metabolites in the correlation networks with those in established metabolic networks derived from genome databases. We find that network similarity indeed decreases with an increasing time difference between these networks during a day/night course and, counter intuitively, that proximity of metabolites in the correlation network is no indicator of proximity of the metabolites in the metabolic network. CONCLUSION: The organizing principles of correlation networks are distinct from those of metabolic reaction maps. Time courses of correlation networks may in the future prove an important data source for understanding these organizing principles.

Regularizing capacity of metabolic networks.

Despite their topological complexity almost all functional properties of metabolic networks can be derived from steady-state dynamics. Indeed, many theoretical investigations (like flux-balance analysis) rely on extracting function from steady states. This leads to the interesting question as to how metabolic networks avoid complex dynamics and maintain a steady-state behavior. Here, we expose metabolic network topologies to binary dynamics generated by simple local rules. We find that the networks' response is highly specific: Complex dynamics are systematically reduced on metabolic networks compared to randomized networks with identical degree sequences. Already small topological modifications substantially enhance the capacity of a network to host complex dynamic behavior and thus reduce its regularizing potential. This exceptionally pronounced regularization of dynamics encoded in the topology may explain why steady-state behavior is ubiquitous in metabolism.

Topology regulates the distribution pattern of excitations in excitable dynamics on graphs.

We study the average excitation density in a simple model of excitable dynamics on graphs and find that this density strongly depends on certain topological features of the graph, namely connectivity and degree correlations, but to a lesser extent on the degree distribution. Remarkably, the average excitation density is changed via the distribution pattern of excitations: An increase in connectivity induces a transition from globally to locally organized excitations and, as a result, leads to an increase in the excitation density. A similar transition can be induced by increasing the rate of spontaneous excitations while keeping the graph architecture constant.