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Maize (Zea mays L.) is one of the three major cereal crops in the world. Leaf angle is an important architectural trait of crops due to its substantial role in light interception by the canopy and hence photosynthetic efficiency. Traditionally, leaf angle has been measured using a protractor, a process that is both slow and laborious. Efficiently measuring leaf angle under field conditions via imaging is challenging due to leaf density in the canopy and the resulting occlusions. However, advances in imaging technologies and machine learning have provided new tools for image acquisition and analysis that could be used to characterize leaf angle using three-dimensional (3D) models of field-grown plants. In this study, PhenoBot 3.0, a robotic vehicle designed to traverse between pairs of agronomically spaced rows of crops, was equipped with multiple tiers of PhenoStereo cameras to capture side-view images of maize plants in the field. PhenoStereo is a customized stereo camera module with integrated strobe lighting for high-speed stereoscopic image acquisition under variable outdoor lighting conditions. An automated image processing pipeline (AngleNet) was developed to measure leaf angles of nonoccluded leaves. In this pipeline, a novel representation form of leaf angle as a triplet of keypoints was proposed. The pipeline employs convolutional neural networks to detect each leaf angle in two-dimensional images and 3D modeling approaches to extract quantitative data from reconstructed models. Our study demonstrates the feasibility of using stereo vision to investigate the distribution of leaf angles in maize under field conditions. The proposed system is an efficient alternative to traditional leaf angle phenotyping and thus could accelerate breeding for improved plant architecture. 

Brassinosteroids (BRs) are a group of plant steroid hormones involved in regulating growth, development, and stress responses. Many components of the BR pathway have previously been identified and characterized. However, BR phenotyping experiments are typically performed on petri plates and/or in a low-throughput manner. Additionally, the BR pathway has extensive crosstalk with drought responses, but drought experiments are time-consuming and difficult to control. Thus, we developed Robotic Assay for Drought (RoAD) to perform BR and drought response experiments in soil-grown Arabidopsis plants. RoAD is equipped with a bench scale, a precisely controlled watering system, an RGB camera, and a laser profilometer. It performs daily weighing, watering, and imaging tasks and is capable of administering BR response assays by watering plants with Propiconazole (PCZ), a BR biosynthesis inhibitor. We developed image processing algorithms for both plant segmentation and phenotypic trait extraction in order to accurately measure traits in 2-dimensional (2D) and 3-dimensional (3D) spaces including plant surface area, leaf length, and leaf width. We then applied machine learning algorithms that utilized the extracted phenotypic parameters to identify image-derived traits that can distinguish control, drought, and PCZ-treated plants. We carried out PCZ and drought experiments on a set of BR mutants and Arabidopsis accessions with altered BR responses. Finally, we extended the RoAD assays to perform BR response assays using PCZ in Zea mays (maize) plants. This study establishes an automated and non-invasive robotic imaging system as a tool to accurately measure morphological and growth-related traits of Arabidopsis and maize plants, providing insights into the BR-mediated control of plant growth and stress responses.