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Abstract Accurate 3D reconstruction is essential for high-throughput plant phenotyping, particularly for studying complex structures such as root systems. While photogrammetry and Structure from Motion (SfM) techniques have become widely used for 3D root imaging, the camera settings used are often underreported in studies, and the impact of camera calibration on model accuracy remains largely underexplored in plant science. In this study, we systematically evaluate the effects of focus, aperture, exposure time, and gain settings on the quality of 3D root models made with a multi-camera scanning system. We show through a series of experiments that calibration significantly improves model quality, with focus misalignment and shallow depth of field (DoF) being the most important factors affecting reconstruction accuracy. Our results further show that proper calibration has a greater effect on reducing noise than filtering it during post-processing, emphasizing the importance of optimizing image acquisition rather than relying solely on computational corrections. This work improves the repeatability and accuracy of 3D root phenotyping by giving useful calibration guidelines. This leads to better trait quantification for use in crop research and plant breeding.more » « lessFree, publicly-accessible full text available March 13, 2026
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Challenge: Digital Imaging of root traits 3D (DIRT/3D) [1] is a software to measure 3D root traits on excavated roots crowns from the field. However, quantifying 3D root traits remains a challenge due to the unknown tradeoff between 3D root-model quality and 3D root-trait accuracy [2]. Questions: Can the 3D root model reconstruction be improved while reducing the image-capturing effort? Does improved 3D root model quality increase the accuracy of trait measurements? Evaluation: Compare reconstruction performance of five open-source 3D model reconstruction pipelines on 12 architecturally contrasting genotypes [1] of field-grown maize roots. Evaluate the accuracy of 3D root traits between the original implementation of DIRT/3D based on VisualSFM with an implementation based on COLMAP. Conclusion: The updated DIRT/3D (COLMAP) pipeline enables quicker image collection by reducing the number of images needed and reducing the human factor during image collection. The results demonstrate that the accuracy of 3D root-trait measurements remained uncompromised.more » « less
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Challenge : Most plant imaging systems focus predominantly on monitoring morphological traits. The challenge is to relate color information to measurements of physiological processes. Question: Can the color of individual leaves be measured and quantified over time to infer physiological information about the plant? Solution: We developed the open source and affordable plant phenotyping software pipeline for Arabidopsis thaliana. SMART (Speedy Measurement of Arabidopsis Rosette Traits) that integrates a new color analysis algorithm to measure leaf surface temperature, leaf wilting and zinc toxicity over time. Data Collection: We used public datasets to develop the algorithm [1] and validate morphological measurements. We also collected top-view images of the Arabidopsis rosette with the Open-Leafmore » « less
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Bucksch, Alexander Clarke (Ed.)Understanding root traits is essential to improve water uptake, increase nitrogen capture and accelerate carbon sequestration from the atmosphere. High-throughput phenotyping to quantify root traits for deeper field-grown roots remains a challenge, however. Recently developed open-source methods use 3D reconstruction algorithms to build 3D models of plant roots from multiple 2D images and can extract root traits and phenotypes. Most of these methods rely on automated image orientation (Structure from Motion)[1] and dense image matching (Multiple View Stereo) algorithms to produce a 3D point cloud or mesh model from 2D images. Until now the performance of these methods when applied to field-grown roots has not been compared tested commonly used open-source pipelines on a test panel of twelve contrasting maize genotypes grown in real field conditions[2-6]. We compare the 3D point clouds produced in terms of number of points, computation time and model surface density. This comparison study provides insight into the performance of different open-source pipelines for maize root phenotyping and illuminates trade-offs between 3D model quality and performance cost for future high-throughput 3D root phenotyping. DOI recognition was not working: https://doi.org/10.1002/essoar.10508794.2more » « less
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Abstract The development of crops with deeper roots holds substantial promise to mitigate the consequences of climate change. Deeper roots are an essential factor to improve water uptake as a way to enhance crop resilience to drought, to increase nitrogen capture, to reduce fertilizer inputs, and to increase carbon sequestration from the atmosphere to improve soil organic fertility. A major bottleneck to achieving these improvements is high-throughput phenotyping to quantify root phenotypes of field-grown roots. We address this bottleneck with Digital Imaging of Root Traits (DIRT)/3D, an image-based 3D root phenotyping platform, which measures 18 architecture traits from mature field-grown maize (Zea mays) root crowns (RCs) excavated with the Shovelomics technique. DIRT/3D reliably computed all 18 traits, including distance between whorls and the number, angles, and diameters of nodal roots, on a test panel of 12 contrasting maize genotypes. The computed results were validated through comparison with manual measurements. Overall, we observed a coefficient of determination of r2>0.84 and a high broad-sense heritability of Hmean2> 0.6 for all but one trait. The average values of the 18 traits and a developed descriptor to characterize complete root architecture distinguished all genotypes. DIRT/3D is a step toward automated quantification of highly occluded maize RCs. Therefore, DIRT/3D supports breeders and root biologists in improving carbon sequestration and food security in the face of the adverse effects of climate change.more » « less
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Abstract Understanding three‐dimensional (3D) root traits is essential to improve water uptake, increase nitrogen capture, and raise carbon sequestration from the atmosphere. However, quantifying 3D root traits by reconstructing 3D root models for deeper field‐grown roots remains a challenge due to the unknown tradeoff between 3D root‐model quality and 3D root‐trait accuracy. Therefore, we performed two computational experiments. We first compared the 3D model quality generated by five state‐of‐the‐art open‐source 3D model reconstruction pipelines on 12 contrasting genotypes of field‐grown maize roots. These pipelines included COLMAP, COLMAP+PMVS (Patch‐based Multi‐View Stereo), VisualSFM, Meshroom, and OpenMVG+MVE (Multi‐View Environment). The COLMAP pipeline achieved the best performance regarding 3D model quality versus computational time and image number needed. In the second test, we compared the accuracy of 3D root‐trait measurement generated by the Digital Imaging of Root Traits 3D pipeline (DIRT/3D) using COLMAP‐based 3D reconstruction with our current DIRT/3D pipeline that uses a VisualSFM‐based 3D reconstruction on the same dataset of 12 genotypes, with 5–10 replicates per genotype. The results revealed that (1) the average number of images needed to build a denser 3D model was reduced from 3000 to 3600 (DIRT/3D [VisualSFM‐based 3D reconstruction]) to around 360 for computational test 1, and around 600 for computational test 2 (DIRT/3D [COLMAP‐based 3D reconstruction]); (2) denser 3D models helped improve the accuracy of the 3D root‐trait measurement; (3) reducing the number of images can help resolve data storage problems. The updated DIRT/3D (COLMAP‐based 3D reconstruction) pipeline enables quicker image collection without compromising the accuracy of 3D root‐trait measurements.more » « less
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SUMMARY The first draft of the Arabidopsis genome was released more than 20 years ago and despite intensive molecular research, more than 30% of Arabidopsis genes remained uncharacterized or without an assigned function. This is in part due to gene redundancy within gene families or the essential nature of genes, where their deletion results in lethality (i.e., thedark genome). High‐throughput plant phenotyping (HTPP) offers an automated and unbiased approach to characterize subtle or transient phenotypes resulting from gene redundancy or inducible gene silencing; however, access to commercial HTPP platforms remains limited. Here we describe the design and implementation ofOPEN leaf, an open‐source phenotyping system with cloud connectivity and remote bilateral communication to facilitate data collection, sharing and processing.OPEN leaf, coupled with our SMART imaging processing pipeline was able to consistently document and quantify dynamic changes at the whole rosette level and leaf‐specific resolution when plants experienced changes in nutrient availability. Our data also demonstrate that VIS sensors remain underutilized and can be used in high‐throughput screens to identify and characterize previously unidentified phenotypes in a leaf‐specific time‐dependent manner. Moreover, the modular and open‐source design ofOPEN leafallows seamless integration of additional sensors based on users and experimental needs.more » « less
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Systems for collecting image data in conjunction with computer vision techniques are a powerful tool for increasing the temporal resolution at which plant phenotypes can be measured non-destructively. Computational tools that are flexible and extendable are needed to address the diversity of plant phenotyping problems. We previously described the Plant Computer Vision (PlantCV) software package, which is an image processing toolkit for plant phenotyping analysis. The goal of the PlantCV project is to develop a set of modular, reusable, and repurposable tools for plant image analysis that are open-source and community-developed. Here we present the details and rationale for major developments in the second major release of PlantCV. In addition to overall improvements in the organization of the PlantCV project, new functionality includes a set of new image processing and normalization tools, support for analyzing images that include multiple plants, leaf segmentation, landmark identification tools for morphometrics, and modules for machine learning.more » « less
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