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Even as the study of plant genomics rapidly develops through the use of high-throughput sequencing techniques, traditional plant phenotyping lags far behind. Here we develop a high-throughput rice phenotyping facility (HRPF) to monitor 13 traditional agronomic traits and 2 newly defined traits during the rice growth period. Using genome-wide association studies (GWAS) of the 15 traits, we identify 141 associated loci, 25 of which contain known genes such as the Green Revolution semi-dwarf gene, SD1 . Based on a performance evaluation of the HRPF and GWAS results, we demonstrate that high-throughput phenotyping has the potential to replace traditional phenotyping techniques and can provide valuable gene identification information. The combination of the multifunctional phenotyping tools HRPF and GWAS provides deep insights into the genetic architecture of important traits. Next-generation sequencing technology has made the generation of huge amounts of genetic data possible, but phenotype characterization remains slow and difficult. Here the authors develop a high-throughput phenotyping facility for rice that is able to accurately identify and characterize traits related to morphology, biomass and yield.

Cereals are the main food for mankind. The grain shape extraction and filled/unfilled grain recognition are meaningful for crop breeding and genetic analysis. The conventional measuring method is mainly manual, which is inefficient, labor-intensive and subjective. Therefore, a novel method was proposed to extract the phenotypic traits of cereal grains based on point clouds. First, a structured light scanner was used to obtain the grains point cloud data. Then, the single grain segmentation was accomplished by image preprocessing, plane fitting, region growth clustering. The length, width, thickness, surface area and volume was calculated by the specified analysis algorithms for grain point cloud. To demonstrate this method, experimental materials included rice, wheat and corn were tested. Compared with manual measurement results, the average measurement error of grain length, width and thickness was 2.07%, 0.97%, 1.13%, and the average measurement efficiency was about 9.6 s per grain. In addition, the grain identification model was conducted with 25 grain phenotypic traits, using 6 machine learning methods. The results showed that the best accuracy for filled/unfilled grain classification was 90.184%.The best accuracy for indica and japonica identification was 99.950%, while for different varieties identification was only 47.252%. Therefore, this method was proved to be an efficient and effective way for crop research.

With increasing demand for novel traits in crop breeding, the plant research community faces the challenge of quantitatively analyzing the structure and function of large numbers of plants. A clear goal of high-throughput phenotyping is to bridge the gap between genomics and phenomics. In this study, we quantified 106 traits from a maize (Zea mays) recombinant inbred line population (n = 167) across 16 developmental stages using the automatic phenotyping platform. Quantitative trait locus (QTL) mapping with a high-density genetic linkage map, including 2,496 recombinant bins, was used to uncover the genetic basis of these complex agronomic traits, and 988 QTLs have been identified for all investigated traits, including three QTL hotspots. Biomass accumulation and final yield were predicted using a combination of dissected traits in the early growth stage. These results reveal the dynamic genetic architecture of maize plant growth and enhance ideotype-based maize breeding and prediction.

Background Rice panicle phenotyping is important in rice breeding, and rice panicle segmentation is the first and key step for image-based panicle phenotyping. Because of the challenge of illumination differentials, panicle shape deformations, rice accession variations, different reproductive stages and the field’s complex background, rice panicle segmentation in the field is a very large challenge. Results In this paper, we propose a rice panicle segmentation algorithm called Panicle-SEG, which is based on simple linear iterative clustering superpixel regions generation, convolutional neural network classification and entropy rate superpixel optimization. To build the Panicle-SEG-CNN model and test the segmentation effects, 684 training images and 48 testing images were randomly selected, respectively. Six indicators, including Qseg, Sr, SSIM, Precision, Recall and F-measure, are employed to evaluate the segmentation effects, and the average segmentation results for the 48 testing samples are 0.626, 0.730, 0.891, 0.821, 0.730, and 76.73%, respectively. Compared with other segmentation approaches, including HSeg, i2 hysteresis thresholding and jointSeg, the proposed Panicle-SEG algorithm has better performance on segmentation accuracy. Meanwhile, the executing speed is also improved when combined with multithreading and CUDA parallel acceleration. Moreover, Panicle-SEG was demonstrated to be a robust segmentation algorithm, which can be expanded for different rice accessions, different field environments, different camera angles, different reproductive stages, and indoor rice images. The testing dataset and segmentation software are available online. Conclusions In conclusion, the results demonstrate that Panicle-SEG is a robust method for panicle segmentation, and it creates a new opportunity for nondestructive yield estimation.

Background Drought threatens the food supply of the world population. Dissecting the dynamic responses of plants to drought will be beneficial for breeding drought-tolerant crops, as the genetic controls of these responses remain largely unknown. Results Here we develop a high-throughput multiple optical phenotyping system to noninvasively phenotype 368 maize genotypes with or without drought stress over a course of 98 days, and collected multiple optical images, including color camera scanning, hyperspectral imaging, and X-ray computed tomography images. We develop high-throughput analysis pipelines to extract image-based traits (i-traits). Of these i-traits, 10,080 were effective and heritable indicators of maize external and internal drought responses. An i-trait-based genome-wide association study reveals 4322 significant locus-trait associations, representing 1529 quantitative trait loci (QTLs) and 2318 candidate genes, many that co-localize with previously reported maize drought responsive QTLs. Expression QTL (eQTL) analysis uncovers many local and distant regulatory variants that control the expression of the candidate genes. We use genetic mutation analysis to validate two new genes, ZmcPGM2 and ZmFAB1A , which regulate i-traits and drought tolerance. Moreover, the value of the candidate genes as drought-tolerant genetic markers is revealed by genome selection analysis, and 15 i-traits are identified as potential markers for maize drought tolerance breeding. Conclusion Our study demonstrates that combining high-throughput multiple optical phenotyping and GWAS is a novel and effective approach to dissect the genetic architecture of complex traits and clone drought-tolerance associated genes.

Nondestructive detection of crop phenotypic traits in the field is very important for crop breeding. Ground-based mobile platforms equipped with sensors can efficiently and accurately obtain crop phenotypic traits. In this study, we propose a dynamic 3D data acquisition method in the field suitable for various crops by using a consumer-grade RGB-D camera installed on a ground-based movable platform, which can collect RGB images as well as depth images of crop canopy sequences dynamically. A scale-invariant feature transform (SIFT) operator was used to detect adjacent date frames acquired by the RGB-D camera to calculate the point cloud alignment coarse matching matrix and the displacement distance of adjacent images. The data frames used for point cloud matching were selected according to the calculated displacement distance. Then, the colored ICP (iterative closest point) algorithm was used to determine the fine matching matrix and generate point clouds of the crop row. The clustering method was applied to segment the point cloud of each plant from the crop row point cloud, and 3D phenotypic traits, including plant height, leaf area and projected area of individual plants, were measured. We compared the effects of LIDAR and image-based 3D reconstruction methods, and experiments were carried out on corn, tobacco, cottons and Bletilla striata in the seedling stage. The results show that the measurements of the plant height (R²= 0.9~0.96, RSME = 0.015~0.023 m), leaf area (R²= 0.8~0.86, RSME = 0.0011~0.0041 ) and projected area (R² = 0.96~0.99) have strong correlations with the manual measurement results. Additionally, 3D reconstruction results with different moving speeds and times throughout the day and in different scenes were also verified. The results show that the method can be applied to dynamic detection with a moving speed up to 0.6 m/s and can achieve acceptable detection results in the daytime, as well as at night. Thus, the proposed method can improve the efficiency of individual crop 3D point cloud data extraction with acceptable accuracy, which is a feasible solution for crop seedling 3D phenotyping outdoors.

Panicle number is directly related to rice yield, so panicle detection and counting has always been one of the most important scientific research topics. Panicle counting is a challenging task due to many factors such as high density, high occlusion, and large variation in size, shape, posture et.al. Deep learning provides state-of-the-art performance in object detection and counting. Generally, the large images need to be resized to fit for the video memory. However, small panicles would be missed if the image size of the original field rice image is extremely large. In this paper, we proposed a rice panicle detection and counting method based on deep learning which was especially designed for detecting rice panicles in rice field images with large image size. Different object detectors were compared and YOLOv5 was selected with MAPE of 3.44% and accuracy of 92.77%. Specifically, we proposed a new method for removing repeated detections and proved that the method outperformed the existing NMS methods. The proposed method was proved to be robust and accurate for counting panicles in field rice images of different illumination, rice accessions, and image input size. Also, the proposed method performed well on UAV images. In addition, an open-access and user-friendly web portal was developed for rice researchers to use the proposed method conveniently.

Summary Drought resistance (DR) is a complex trait that is regulated by a variety of genes. Without comprehensive profiling of DR‐related traits, the knowledge of the genetic architecture for DR in cotton remains limited. Thus, there is a need to bridge the gap between genomics and phenomics. In this study, an automatic phenotyping platform (APP) was systematically applied to examine 119 image‐based digital traits (i‐traits) during drought stress at the seedling stage, across a natural population of 200 representative upland cotton accessions. Some novel i‐traits, as well as some traditional i‐traits, were used to evaluate the DR in cotton. The phenomics data allowed us to identify 390 genetic loci by genome‐wide association study (GWAS) using 56 morphological and 63 texture i‐traits. DR‐related genes, including GhRD2, GhNAC4, GhHAT22 and GhDREB2, were identified as candidate genes by some digital traits. Further analysis of candidate genes showed that Gh_A04G0377 and Gh_A04G0378 functioned as negative regulators for cotton drought response. Based on the combined digital phenotyping, GWAS analysis and transcriptome data, we conclude that the phenomics dataset provides an excellent resource to characterize key genetic loci with an unprecedented resolution which can inform future genome‐based breeding for improved DR in cotton.

China is the largest producer and consumer of rice, and the classification of filled/unfilled rice grains is of great significance for rice breeding and genetic analysis. The traditional method for filled/unfilled rice grain identification was generally manual, which had the disadvantages of low efficiency, poor repeatability, and low precision. In this study, we have proposed a novel method for filled/unfilled grain classification based on structured light imaging and Improved PointNet++. Firstly, the 3D point cloud data of rice grains were obtained by structured light imaging. And then the specified processing algorithms were developed for the single grain segmentation, and data enhancement with normal vector. Finally, the PointNet++ network was improved by adding an additional Set Abstraction layer and combining the maximum pooling of normal vectors to realize filled/unfilled rice grain point cloud classification. To verify the model performance, the Improved PointNet++ was compared with six machine learning methods, PointNet and PointConv. The results showed that the optimal machine learning model is XGboost, with a classification accuracy of 91.99%, while the classification accuracy of Improved PointNet++ was 98.50% outperforming the PointNet 93.75% and PointConv 92.25%. In conclusion, this study has demonstrated a novel and effective method for filled/unfilled grain recognition.

Manual phenotyping of rice tillers is time consuming and labor intensive, and lags behind the rapid development of rice functional genomics. Thus, automated, non-destructive methods of phenotyping rice tiller traits at a high spatial resolution and high throughput for large-scale assessment of rice accessions are urgently needed. In this study, we developed a high-throughput micro-CT-RGB imaging system to non-destructively extract 739 traits from 234 rice accessions at nine time points. We could explain 30% of the grain yield variance from two tiller traits assessed in the early growth stages. A total of 402 significantly associated loci were identified by genome-wide association study, and dynamic and static genetic components were found across the nine time points. A major locus associated with tiller angle was detected at time point 9, which contained a major gene, TAC1. Significant variants associated with tiller angle were enriched in the 3ʹ-untranslated region of TAC1. Three haplotypes for the gene were found, and rice accessions containing haplotype H3 displayed much smaller tiller angles. Further, we found two loci containing associations with both vigor-related traits identified by high-throughput micro-CT-RGB imaging and yield. The superior alleles would be beneficial for breeding for high yield and dense planting.