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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. 

Abstract Optimizing leaf angle and other canopy architecture traits has helped modern maize (Zea maysL.) become adapted to higher planting densities over the last 60 years. Traditional investigations into genetic control of leaf angle have focused on one leaf or the average of multiple leaves; as a result, our understanding of genetic control across multiple canopy levels is still limited. To address this, genetic mapping across four canopy levels was conducted in the present study to investigate the genetic control of leaf angle across the canopy. We developed two populations of doubled haploid lines derived from three inbreds with distinct leaf angle phenotypes. These populations were genotyped with genotyping‐by‐sequencing and phenotyped for leaf angle at four different canopy levels over multiple years. To understand how leaf angle changes across the canopy, the four measurements were used to derive three additional traits. Composite interval mapping was conducted with the leaf‐specific measurements and the derived traits. A set of 59 quantitative trait loci (QTLs) were uncovered for seven traits, and two genomic regions were consistently detected across multiple canopy levels. Additionally, seven genomic regions were found to contain consistent QTLs with either relatively stable or dynamic effects at different canopy levels. Prioritizing the selection of QTLs with dynamic effects across the canopy will aid breeders in selecting maize hybrids with the ideal canopy architecture that continues to maximize yield on a per area basis under increasing planting densities. 

We report de novo genome assemblies, transcriptomes, annotations, and methylomes for the 26 inbreds that serve as the founders for the maize nested association mapping population. The number of pan-genes in these diverse genomes exceeds 103,000, with approximately a third found across all genotypes. The results demonstrate that the ancient tetraploid character of maize continues to degrade by fractionation to the present day. Excellent contiguity over repeat arrays and complete annotation of centromeres revealed additional variation in major cytological landmarks. We show that combining structural variation with single-nucleotide polymorphisms can improve the power of quantitative mapping studies. We also document variation at the level of DNA methylation and demonstrate that unmethylated regions are enriched for cis-regulatory elements that contribute to phenotypic variation.