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Societal Impact StatementPlant breeding is a critical tool for increasing the productivity, climate resilience, and sustainability of agriculture, but current phenotyping methods are a bottleneck due to the amount of human labor involved. Here, we demonstrate high‐throughput phenotyping with an unmanned aerial vehicle (UAV) to analyze the season‐long flowering pattern in cotton, subsequently mapping relevant genetic factors underpinning the trait. Season‐long flowering is a complex trait, with implications for adaptation of perennials to specific environments. We believe our approach can improve the speed and efficacy of breeding for a variety of woody perennials. SummaryMany perennial plants make important contributions to agroeconomies and agroecosystems but have complex architecture and/or long flowering duration that hinders measurement and selection. Iteratively tracking productivity over a long flowering/fruiting season may permit the identification of genetic factors conferring different reproductive strategies that might be successful in different environments, ranging from rapid early maturation that avoids stresses, to late maturation that utilizes the full seasonal duration to maximize productivity.In cotton, a perennial plant that is generally cultivated as an annual crop, we apply aerial imagery and deep learning methods to novel and stable genetic stocks, identifying genetic factors influencing the duration and rate of fruiting.Our phenotyping method was able to identify 24 QTLs that affect flowering behavior in cotton. A total of five of these corresponded to previously identified QTLs from other studies.While these factors may have different relationships with crop productivity and quality in different environments, their determination adds potentially important information to breeding decisions. With transfer learning of the deep learning models, this approach could be applied widely, potentially improving gains from selection in diverse perennial shrubs and trees essential to sustainable agricultural intensification. 

Abstract BackgroundPlant architecture can influence crop yield and quality. Manual extraction of architectural traits is, however, time-consuming, tedious, and error prone. The trait estimation from 3D data addresses occlusion issues with the availability of depth information while deep learning approaches enable learning features without manual design. The goal of this study was to develop a data processing workflow by leveraging 3D deep learning models and a novel 3D data annotation tool to segment cotton plant parts and derive important architectural traits. ResultsThe Point Voxel Convolutional Neural Network (PVCNN) combining both point- and voxel-based representations of 3D data shows less time consumption and better segmentation performance than point-based networks. Results indicate that the best mIoU (89.12%) and accuracy (96.19%) with average inference time of 0.88 s were achieved through PVCNN, compared to Pointnet and Pointnet++. On the seven derived architectural traits from segmented parts, an R²value of more than 0.8 and mean absolute percentage error of less than 10% were attained. ConclusionThis plant part segmentation method based on 3D deep learning enables effective and efficient architectural trait measurement from point clouds, which could be useful to advance plant breeding programs and characterization of in-season developmental traits. The plant part segmentation code is available athttps://github.com/UGA-BSAIL/plant_3d_deep_learning. 

Abstract Yield improvement in cotton could be accelerated through selection for functional yield drivers such as interception of cumulative photosynthetically active radiation (∑IPAR), radiation use efficiency (RUE), and harvest index (HI). However, information on the extent to which these traits vary in cotton in the southeastern United States is limited. It was hypothesized that functional yield drivers would vary significantly within a diverse cotton collection. This study was conducted in Tifton and Athens, GA, and included a total of 4 site‐years. Lint yield, total biomass production, ∑IPAR, RUE, and HI were all affected by genotype. Biomass was more strongly correlated with RUE than ∑IPAR. Even among the highest yielding genotypes, values for functional yield drivers (biomass and harvest index) differed significantly, indicating that high yields could be achieved by differentially manipulating these underlying traits. However, when considered for all genotypes, only HI exhibited a significant positive correlation with yield. Boll production and intra‐boll yield components were also affected by genotype. When considered across upland genotypes, lint per boll, lint per seed, and lint percent were strongly associated with HI and lint yield, whereas boll mass and seed number per boll were not. We conclude that the genotypes evaluated in the current study achieve high lint production per boll and lint yields by manipulating different yield drivers. However, lint yield was primarily maximized through an increase in HI due to increases in boll production and within‐boll distribution of biomass to fiber, not due to increases in total biomass production or boll size. 

Understanding the complex interactions between genotype-environment dynamics is fundamental for optimizing crop improvement. However, traditional phenotyping methods limit assessments to the end of the growing season, restricting continuous crop monitoring. To address this limitation, we developed a methodology for spatiotemporal registration of time-series 3D point cloud data, enabling field phenotyping over time for accurate crop growth tracking. Leveraging multi-scan terrestrial laser scanning (TLS), we captured high-resolution 3D LiDAR data in a cotton breeding field across various stages of the growing season to generate four-dimensional (4D) crop models, seamlessly integrating spatial and temporal dimensions. Our registration procedure involved an initial pairwise terrain-based matching for rough alignment, followed by a bird’s-eye view adjustment for fine registration. Point clouds collected throughout nine sessions across the growing season were successfully registered both spatially and temporally, with average registration errors of approximately 3 cm. We used the generated 4D models to monitor canopy height (CH) and volume (CV) for eleven cotton genotypes over two months. The consistent height reference established via our spatiotemporal registration process enabled precise estimations of CH (R² = 0.95, RMSE = 7.6 cm). Additionally, we analyzed the relationship between CV and the interception of photosynthetically active radiation (IPAR_f), finding that it followed a curve with exponential saturation, consistent with theoretical models, with a standard error of regression (SER) of 11%. In addition, we compared mathematical models from the Richards family of sigmoid curves for crop growth modeling, finding that the logistic model effectively captured CH and CV evolution, aiding in identifying significant genotype differences. Our novel TLS-based digital phenotyping methodology enhances precision and efficiency in field phenotyping over time, advancing plant phenomics and empowering efficient decision-making for crop improvement efforts. 

Cotton breeding programs have focused on agronomically-desirable traits. Without targeted selection for tolerance to high temperature extremes, cotton will likely be more vulnerable to environment-induced yield loss. Recently-developed methods that couple chlorophyll fluorescence induction measurements with temperature response experiments could be used to identify genotypic variation in photosynthetic thermotolerance of specific photosynthetic processes for field-grown plants. It was hypothesized that diverse cotton genotypes would differ significantly in photosynthetic thermotolerance, specific thylakoid processes would exhibit differential sensitivities to high temperature, and that the most heat tolerant process would exhibit substantial genotypic variation in thermotolerance plasticity. A two-year field experiment was conducted at Tifton and Athens, Georgia, USA. Experiments included 10 genotypes in 2020 and 11 in 2021. Photosynthetic thermotolerance for field-collected leaf samples was assessed by determining the high temperature threshold resulting in a 15% decline in photosynthetic efficiency (T15) for energy trapping by photosystem II (ΦPo), intersystem electron transport (ΦEo), and photosystem I end electron acceptor reduction (ΦRo). Significant genotypic variation in photosynthetic thermotolerance was observed, but the response was dependent on location and photosynthetic parameter assessed. ΦEo was substantially more heat sensitive than ΦPo or ΦRo. Significant genotypic variation in thermotolerance plasticity of ΦEo was also observed. Identifying the weakest link in photosynthetic tolerance to high temperature will facilitate future selection efforts by focusing on the most heat-susceptible processes. Given the genotypic differences in environmental plasticity observed here, future research should evaluate genotypic variation in acclimation potential in controlled environments. 

Estimation of cotton yield before harvest offers many benefits to breeding programs, researchers and producers. Remote sensing enables efficient and consistent estimation of cotton yields, as opposed to traditional field measurements and surveys. The overall goal of this study was to develop a data processing pipeline to perform fast and accurate pre-harvest yield predictions of cotton breeding fields from aerial imagery using machine learning techniques. By using only a single plot image extracted from an orthomosaic map, a Support Vector Machine (SVM) classifier with four selected features was trained to identify the cotton pixels present in each plot image. The SVM classifier achieved an accuracy of 89%, a precision of 86%, a recall of 75%, and an F1-score of 80% at recognizing cotton pixels. After performing morphological image processing operations and applying a connected components algorithm, the classified cotton pixels were clustered to predict the number of cotton bolls at the plot level. Our model fitted the ground truth counts with anR²value of 0.93, a normalized root mean squared error of 0.07, and a mean absolute percentage error of 13.7%. This study demonstrates that aerial imagery with machine learning techniques can be a reliable, efficient, and effective tool for pre-harvest cotton yield prediction.