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.

Leaves are the plant’s solar panel and food factory, and leaf traits are always key issues to investigate in plant research. Traditional methods for leaf trait measurement are time-consuming. In this work, an engineering prototype has been established for high-throughput leaf scoring (HLS) of a large number of Oryza sativa accessions. The mean absolute per cent of errors in traditional measurements versus HLS were below 5% for leaf number, area, shape, and colour. Moreover, HLS can measure up to 30 leaves per minute. To demonstrate the usefulness of HLS in dissecting the genetic bases of leaf traits, a genome-wide association study (GWAS) was performed for 29 leaf traits related to leaf size, shape, and colour at three growth stages using HLS on a panel of 533 rice accessions. Nine associated loci contained known leaf-related genes, such as Nal1 for controlling the leaf width. In addition, a total of 73, 123, and 177 new loci were detected for traits associated with leaf size, colour, and shape, respectively. In summary, after evaluating the performance with a large number of rice accessions, the combination of GWAS and high-throughput leaf phenotyping (HLS) has proven a valuable strategy to identify the genetic loci controlling rice leaf traits.

Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack a direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High-quality 3D volume data from our enhanced methods facilitate sophisticated and effective computational segmentation. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high-resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.

The fitness of the rape leaf is closely related to its biomass and photosynthesis. The study of leaf traits is significant for improving rape leaf production and optimizing crop management. Canopy structure and individual leaf traits are the major indicators of quality during the rape seedling stage. Differences in canopy structure reflect the influence of environmental factors such as water, sunlight and nutrient supply. The traits of individual rape leaves traits indicate the growth period of the rape as well as its canopy shape. We established a high-throughput stereo-imaging system for the reconstruction of the three-dimensional canopy structure of rape seedlings from which leaf area and plant height can be extracted. To evaluate the measurement accuracy of leaf area and plant height, 66 rape seedlings were randomly selected for automatic and destructive measurements. Compared with the manual measurements, the mean absolute percentage error of automatic leaf area and plant height measurements was 3.68 and 6.18%, respectively, and the squares of the correlation coefficients (R2) were 0.984 and 0.845, respectively. Compared with the two-dimensional projective imaging method, the leaf area extracted using stereo-imaging was more accurate. In addition, a semi-automatic image analysis pipeline was developed to extract 19 individual leaf shape traits, including 11 scale-invariant traits, 3 inner cavity related traits, and 5 margin-related traits, from the images acquired by the stereo-imaging system. We used these quantified traits to classify rapes according to three different leaf shapes: mosaic-leaf, semi-mosaic-leaf, and round-leaf. Based on testing of 801 seedling rape samples, we found that the leave-one-out cross validation classification accuracy was 94.4, 95.6, and 94.8% for stepwise discriminant analysis, the support vector machine method and the random forest method, respectively. In this study, a nondestructive and high-throughput stereo-imaging system was developed to quantify canopy three-dimensional structure and individual leaf shape traits with improved accuracy, with implications for rape phenotyping, functional genomics, and breeding.

Time-lapse 3D imaging, computer vision, and mathematical modeling were used to quantify the root system architectures of three maize genotypes as they grew in c