Visualizing the structural and chemical heterogeneity of fruit and vegetables using advanced imaging techniques: fundamentals, instrumental aspects, applications and future perspectives.

Fruit and vegetables (F&V) are vastly complicated products with highly diverse chemical and structural characteristics. Advanced imaging techniques either combine imaging with spectral information or can provide excellent tissue penetration, and enable the possibility to target, visualize and even qualify the chemical and physical (structural) heterogeneity within F&V. In this review, visible and/or near infrared hyperspectral imaging, Fourier transform infrared microspectroscopic imaging, Raman imaging, X-ray and magnetic resonance imaging to reveal chemical and structural information in a spatial context of F&V at the macro- (entire products), meso- (tissues), and micro- (individual cells) scales are comprehensively summarized. In addition, their basic concepts and operational procedures, particularly sample preparation and instrumental parameter adjustments, are addressed. Finally, future challenges and perspectives of these techniques are put forward. These imaging techniques are powerful tools to assess the biochemical and structural heterogeneity of F&V. Cost reduction, sensor fusion and data sharing platforms are future trends. More emphasis on aspects of knowledge and extension at the level of academia and research, especially on how to select techniques, choose operational parameters and prepare samples, are important to overcome barriers for the wider adoption of these techniques to improve the evaluation of F&V quality.

Hyperspectral imaging reveals chemical heterogeneity of fruit and vegetables.Imaging techniques provide spatial insights in fruit and vegetables at multiple scales.Future trends are cost reduction, sensor fusion and data sharing.Instrumental adjustment and sample preparation should receive more attention.

New insight into the genetic basis of oil content based on noninvasive three-dimensional phenotyping and tissue-specific transcriptome in Brassica napus.

BACKGROUND: Increasing seed oil content is the most important breeding goal in Brassica napus, and phenotyping is crucial to dissect its genetic basis in crops. To date, QTL mapping for oil content has been based on whole seeds, and the lipid distribution is far from uniform in different tissues of seeds in B. napus. In this case, the phenotype based on whole seeds was unable to sufficiently reveal the complex genetic characteristics of seed oil content. RESULTS: Here, the three-dimensional (3D) distribution of lipid was determined for B. napus seeds by magnetic resonance imaging (MRI) and 3D quantitative analysis, and ten novel oil content-related traits were obtained by subdividing the seeds. Based on a high-density genetic linkage map, 35 QTLs were identified for 4 tissues, the outer cotyledon (OC), inner cotyledon (IC), radicle (R) and seed coat (SC), which explained up to 13.76% of the phenotypic variation. Notably, 14 tissue-specific QTLs were reported for the first time, 7 of which were novel. Moreover, haplotype analysis showed that the favorable alleles for different seed tissues exhibited cumulative effects on oil content. Furthermore, tissue-specific transcriptomes revealed that more active energy and pyruvate metabolism influenced carbon flow in the IC, OC and R than in the SC at the early and middle seed development stages, thus affecting the distribution difference in oil content. Combining tissue-specific QTL mapping and transcriptomics, 86 important candidate genes associated with lipid metabolism were identified that underlie 19 unique QTLs, including the fatty acid synthesis rate-limiting enzyme-related gene CAC2, in the QTLs for OC and IC. CONCLUSIONS: The present study provides further insight into the genetic basis of seed oil content at the tissue-specific level.

Design and Development of a Low-Cost UGV 3D Phenotyping Platform with Integrated LiDAR and Electric Slide Rail.

Unmanned ground vehicles (UGV) have attracted much attention in crop phenotype monitoring due to their lightweight and flexibility. This paper describes a new UGV equipped with an electric slide rail and point cloud high-throughput acquisition and phenotype extraction system. The designed UGV is equipped with an autopilot system, a small electric slide rail, and Light Detection and Ranging (LiDAR) to achieve high-throughput, high-precision automatic crop point cloud acquisition and map building. The phenotype analysis system realized single plant segmentation and pipeline extraction of plant height and maximum crown width of the crop point cloud using the Random sampling consistency (RANSAC), Euclidean clustering, and k-means clustering algorithm. This phenotyping system was used to collect point cloud data and extract plant height and maximum crown width for 54 greenhouse-potted lettuce plants. The results showed that the correlation coefficient (R2) between the collected data and manual measurements were 0.97996 and 0.90975, respectively, while the root mean square error (RMSE) was 1.51 cm and 4.99 cm, respectively. At less than a tenth of the cost of the PlantEye F500, UGV achieves phenotypic data acquisition with less error and detects morphological trait differences between lettuce types. Thus, it could be suitable for actual 3D phenotypic measurements of greenhouse crops.

High-Throughput 3D Phenotyping of Plant Shoot Apical Meristems From Tissue-Resolution Data.

Confocal imaging is a well-established method for investigating plant phenotypes on the tissue and organ level. However, many differences are difficult to assess by visual inspection and researchers rely extensively on ad hoc manual quantification techniques and qualitative assessment. Here we present a method for quantitatively phenotyping large samples of plant tissue morphologies using triangulated isosurfaces. We successfully demonstrate the applicability of the approach using confocal imaging of aerial organs in Arabidopsis thaliana. Automatic identification of flower primordia using the surface curvature as an indication of outgrowth allows for high-throughput quantification of divergence angles and further analysis of individual flowers. We demonstrate the throughput of our method by quantifying geometric features of 1065 flower primordia from 172 plants, comparing auxin transport mutants to wild type. Additionally, we find that a paraboloid provides a simple geometric parameterisation of the shoot inflorescence domain with few parameters. We utilise parameterisation methods to provide a computational comparison of the shoot apex defined by a fluorescent reporter of the central zone marker gene CLAVATA3 with the apex defined by the paraboloid. Finally, we analyse the impact of mutations which alter mechanical properties on inflorescence dome curvature and compare the results with auxin transport mutants. Our results suggest that region-specific expression domains of genes regulating cell wall biosynthesis and local auxin transport can be important in maintaining the wildtype tissue shape. Altogether, our results indicate a general approach to parameterise and quantify plant development in 3D, which is applicable also in cases where data resolution is limited, and cell segmentation not possible. This enables researchers to address fundamental questions of plant development by quantitative phenotyping with high throughput, consistency and reproducibility.

Performances Evaluation of a Low-Cost Platform for High-Resolution Plant Phenotyping.

This study aims to test the performances of a low-cost and automatic phenotyping platform, consisting of a Red-Green-Blue (RGB) commercial camera scanning objects on rotating plates and the reconstruction of main plant phenotypic traits via the structure for motion approach (SfM). The precision of this platform was tested in relation to three-dimensional (3D) models generated from images of potted maize, tomato and olive tree, acquired at a different frequency (steps of 4°, 8° and 12°) and quality (4.88, 6.52 and 9.77 µm/pixel). Plant and organs heights, angles and areas were extracted from the 3D models generated for each combination of these factors. Coefficient of determination (R2), relative Root Mean Square Error (rRMSE) and Akaike Information Criterion (AIC) were used as goodness-of-fit indexes to compare the simulated to the observed data. The results indicated that while the best performances in reproducing plant traits were obtained using 90 images at 4.88 µm/pixel (R2 = 0.81, rRMSE = 9.49% and AIC = 35.78), this corresponded to an unviable processing time (from 2.46 h to 28.25 h for herbaceous plants and olive trees, respectively). Conversely, 30 images at 4.88 µm/pixel resulted in a good compromise between a reliable reconstruction of considered traits (R2 = 0.72, rRMSE = 11.92% and AIC = 42.59) and processing time (from 0.50 h to 2.05 h for herbaceous plants and olive trees, respectively). In any case, the results pointed out that this input combination may vary based on the trait under analysis, which can be more or less demanding in terms of input images and time according to the complexity of its shape (R2 = 0.83, rRSME = 10.15% and AIC = 38.78). These findings highlight the reliability of the developed low-cost platform for plant phenotyping, further indicating the best combination of factors to speed up the acquisition and elaboration process, at the same time minimizing the bias between observed and simulated data.

Three-Dimensional Lesion Phenotyping and Physiologic Characterization Inform Remyelination Ability in Multiple Sclerosis.

BACKGROUND AND PURPOSE: Multiple sclerosis (MS) clinical management is based upon lesion characterization from 2-dimensional (2D) magnetic resonance imaging (MRI) views. Such views fail to convey the lesion-phenotype (ie, shape and surface texture) complexity, underlying metabolic alterations, and remyelination potential. We utilized a 3-dimensional (3D) lesion phenotyping approach coupled with imaging to study physiologic profiles within and around MS lesions and their impacts on lesion phenotypes. METHODS: Lesions were identified in 3T T2 -FLAIR images and segmented using geodesic active contouring. A calibrated fMRI sequence permitted measurement of cerebral blood flow (CBF), blood-oxygen-level-dependent signal (BOLD), and cerebral metabolic rate of oxygen (CMRO2 ). These metrics were measured within lesions and surrounding tissue in concentric layers exact to the 3D-lesion shape. BOLD slope was calculated as BOLD changes from a lesion to its surrounding perimeters. White matter integrity was measured using diffusion kurtosis imaging. Associations between these metrics and 3D-lesion phenotypes were studied. RESULTS: One hundred nine lesions from 23 MS patients were analyzed. We identified a noninvasive biomarker, BOLD slope, to metabolically characterize lesions. Positive BOLD slope lesions were metabolically active with higher CMRO2 and CBF compared to negative BOLD slope or inactive lesions. Metabolically active lesions with more intact white matter integrity had more symmetrical shapes and more complex surface textures compared to inactive lesions with less intact white matter integrity. CONCLUSION: The association of lesion phenotypes with their metabolic signatures suggests the prospect for translation of such data to clinical management by providing information related to metabolic activity, lesion age, and risk for disease reactivation and self-repair. Our findings also provide a platform for disease surveillance and outcome quantification involving myelin repair therapeutics.

LiDARPheno - A Low-Cost LiDAR-Based 3D Scanning System for Leaf Morphological Trait Extraction.

The ever-growing world population brings the challenge for food security in the current world. The gene modification tools have opened a new era for fast-paced research on new crop identification and development. However, the bottleneck in the plant phenotyping technology restricts the alignment in geno-pheno development as phenotyping is the key for the identification of potential crop for improved yield and resistance to the changing environment. Various attempts to making the plant phenotyping a "high-throughput" have been made while utilizing the existing sensors and technology. However, the demand for 'good' phenotypic information for linkage to the genome in understanding the gene-environment interactions is still a bottleneck in the plant phenotyping technologies. Moreover, the available technologies and instruments are inaccessible, expensive, and sometimes bulky. This work attempts to address some of the critical problems, such as exploration and development of a low-cost LiDAR-based platform for phenotyping the plants in-lab and in-field. A low-cost LiDAR-based system design, LiDARPheno, is introduced in this work to assess the feasibility of the inexpensive LiDAR sensor in the leaf trait (length, width, and area) extraction. A detailed design of the LiDARPheno, based on low-cost and off-the-shelf components and modules, is presented. Moreover, the design of the firmware to control the hardware setup of the system and the user-level python-based script for data acquisition is proposed. The software part of the system utilizes the publicly available libraries and Application Programming Interfaces (APIs), making it easy to implement the system by a non-technical user. The LiDAR data analysis methods are presented, and algorithms for processing the data and extracting the leaf traits are developed. The processing includes conversion, cleaning/filtering, segmentation and trait extraction from the LiDAR data. Experiments on indoor plants and canola plants were performed for the development and validation of the methods for estimation of the leaf traits. The results of the LiDARPheno based trait extraction are compared with the SICK LMS400 (a commercial 2D LiDAR) to assess the performance of the developed system.