Digital phenotyping and genotype-to-phenotype (G2P) models to predict complex traits in cereal crops

The revolution in digital phenotyping combined with the new layers of omics and envirotyping tools offers great promise to improve selection and accelerate genetic gains for crop improvement. This chapter examines the latest methods involving digital phenotyping tools to predict complex traits in cereals crops. The chapter has two parts. In the first part, entitled “Digital phenotyping as a tool to support breeding programs”, the secondary phenotypes measured by high-throughput plant phenotyping that are potentially useful for breeding are reviewed. In the second part, “Implementing complex G2P models in breeding programs”, the integration of data from digital phenotyping into genotype to phenotype (G2P) models to improve the prediction of complex traits using genomic information is discussed. The current status of statistical models to incorporate secondary traits in univariate and multivariate models, as well as how to better handle longitudinal (for example light interception, biomass accumulation, canopy height) traits, is reviewe

Row selection in remote sensing from four-row plots of maize and sorghum based on repeatability and predictive modeling

Remote sensing enables the rapid assessment of many traits that provide valuable information to plant breeders throughout the growing season to improve genetic gain. These traits are often extracted from remote sensing data on a row segment (rows within a plot) basis enabling the quantitative assessment of any row-wise subset of plants in a plot, rather than a few individual representative plants, as is commonly done in field-based phenotyping. Nevertheless, which rows to include in analysis is still a matter of debate. The objective of this experiment was to evaluate row selection and plot trimming in field trials conducted using four-row plots with remote sensing traits extracted from RGB (red-green-blue), LiDAR (light detection and ranging), and VNIR (visible near infrared) hyperspectral data. Uncrewed aerial vehicle flights were conducted throughout the growing seasons of 2018 to 2021 with data collected on three years of a sorghum experiment and two years of a maize experiment. Traits were extracted from each plot based on all four row segments (RS) (RS1234), inner rows (RS23), outer rows (RS14), and individual rows (RS1, RS2, RS3, and RS4). Plot end trimming of 40 cm was an additional factor tested. Repeatability and predictive modeling of end-season yield were used to evaluate performance of these methodologies. Plot trimming was never shown to result in significantly different outcomes from non-trimmed plots. Significant differences were often observed based on differences in row selection. Plots with more row segments were often favorable for increasing repeatability, and excluding outer rows improved predictive modeling. These results support long-standing principles of experimental design in agronomy and should be considered in breeding programs that incorporate remote sensing

Moisture content estimation and senescence phenotyping of novel Miscanthus hybrids combining UAV-based remote sensing and machine learning

Miscanthus is a leading perennial biomass crop that can produce high yields on marginal lands. Moisture content is a highly relevant biomass quality trait with multiple impacts on efficiencies of harvest, transport, and storage. The dynamics of moisture content during senescence and overwinter ripening are determined by genotype × environment interactions. In this paper, unmanned aerial vehicle (UAV)-based remote sensing was used for high-throughput plant phenotyping (HTPP) of the moisture content dynamics during autumn and winter senescence of 14 contrasting hybrid types (progeny of M. sinensis x M. sinensis [M. sin x M. sin, eight types] and M. sinensis x M. sacchariflorus [M. sin x M. sac, six types]). The time series of moisture content was estimated using machine learning (ML) models and a range of vegetation indices (VIs) derived from UAV-based remote sensing. The most important VIs for moisture content estimation were selected by the recursive feature elimination (RFE) algorithm and were BNDVI, GDVI, and PSRI. The ML model transferability was high only when the moisture content was above 30%. The best ML model accuracy was achieved by combining VIs and categorical variables (5.6% of RMSE). This model was used for phenotyping senescence dynamics and identifying the stay-green (SG) trait of Miscanthus hybrids using the generalized additive model (GAM). Combining ML and GAM modeling, applied to time series of moisture content values estimated from VIs derived from multiple UAV flights, proved to be a powerful tool for HTPP

High-throughput screening of soybean di-nitrogen fixation and seed nitrogen content using spectral sensing

Symbiotic di-nitrogen fixation of grain legumes has a substantial impact on crop performance, harvest product quality, and nitrogen (N) balance of crop rotations, particularly under organic management regimes. In soybean breeding, selection for increased nitrogen fixation is desirable for improving seed protein content and N balance of cropping systems. However, the lack of high-throughput screening methods for direct measurement of N2 fixation rates prohibits practical breeding efforts. Therefore, hyperspectral canopy reflectance measurement as a field-based phenotyping method was evaluated in three environments for indirect estimation of N fixation and uptake of soil nitrogen in a set of early maturity soybean genotypes exhibiting a wide range in seed protein content. Reflectance spectra were collected in repeated measurements during flowering and early seed filling stages. Subsequently, various spectral reflectance indices (SRIs) were calculated for characterizing nitrogen accumulation of individual genotypes. Moreover, prediction models for seed protein content as an end-of-season target trait were developed utilizing full spectral information in partial-least-square regression (PLSR) models. A number of N-related SRIs calculated from spectral reflectance data recorded at the beginning of the seed filling stage were significantly correlated to seed protein content. The best prediction of seed protein content, however, was achieved in PLSR models (validation R2 = 0.805 across all three environments). Environments lower in initial soil mineral N content appeared as more favorable selection sites in terms of prediction accuracy, because N fixation is not masked by soil N uptake in such environments. Hyperspectral reflectance data proved to be a valuable method for determining genetic variation in crop N accumulation, which might be implemented in highthroughput screening protocols for N fixation in plant breeding programs

Field phenotyping for African crops: overview and perspectives

Improvements in crop productivity are required to meet the dietary demands of the rapidly-increasing African population. The development of key staple crop cultivars that are high-yielding and resilient to biotic and abiotic stresses is essential. To contribute to this objective, high-throughput plant phenotyping approaches are important enablers for the African plant science community to measure complex quantitative phenotypes and to establish the genetic basis of agriculturally relevant traits. These advances will facilitate the screening of germplasm for optimum performance and adaptation to low-input agriculture and resource-constrained environments. Increasing the capacity to investigate plant function and structure through non-invasive technologies is an effective strategy to aid plant breeding and additionally may contribute to precision agriculture. However, despite the significant global advances in basic knowledge and sensor technology for plant phenotyping, Africa still lags behind in the development and implementation of these systems due to several practical, financial, geographical and political barriers. Currently, field phenotyping is mostly carried out by manual methods that are prone to error, costly, labor-intensive and may come with adverse economic implications. Therefore, improvements in advanced field phenotyping capabilities and appropriate implementation are key factors for success in modern breeding and agricultural monitoring. In this review, we provide an overview of the current state of field phenotyping and the challenges limiting its implementation in some African countries. We suggest that the lack of appropriate field phenotyping infrastructures is impeding the development of improved crop cultivars and will have a detrimental impact on the agricultural sector and on food security. We highlight the prospects for integrating emerging and advanced low-cost phenotyping technologies into breeding protocols and characterizing crop responses to environmental challenges in field experimentation. Finally, we explore strategies for overcoming the barriers and maximizing the full potential of emerging field phenotyping technologies in African agriculture. This review paper will open new windows and provide new perspectives for breeders and the entire plant science community in Africa.BBSRC: BB/P016855/

Canopy Roughness: A New Phenotypic Trait to Estimate Aboveground Biomass from Unmanned Aerial System

Cost-effective phenotyping methods are urgently needed to advance crop genetics in order to meet the food, fuel, and fiber demands of the coming decades. Concretely, charac-terizing plot level traits in fields is of particular interest. Re-cent developments in high resolution imaging sensors for UAS (unmanned aerial systems) focused on collecting de-tailed phenotypic measurements are a potential solution. We introduce canopy roughness as a new plant plot-level trait. We tested its usability with soybean by optical data collect-ed from UAS to estimate biomass. We validate canopy roughness on a panel of 108 soybean [Glycine max (L.) Merr.] recombinant inbred lines in a multienvironment trial during the R2 growth stage. A senseFly eBee UAS platform obtained aerial images with a senseFly S.O.D.A. compact digital camera. Using a structure from motion (SfM) tech-nique, we reconstructed 3D point clouds of the soybean experiment. A novel pipeline for feature extraction was de-veloped to compute canopy roughness from point clouds. We used regression analysis to correlate canopy roughness with field-measured aboveground biomass (AGB) with a leave-one-out cross-validation. Overall, our models achieved a coefficient of determination (R2) greater than 0.5 in all trials. Moreover, we found that canopy roughness has the ability to discern AGB variations among different geno-types. Our test trials demonstrate the potential of canopy roughness as a reliable trait for high-throughput phenotyping to estimate AGB. As such, canopy roughness provides practical information to breeders in order to select pheno-types on the basis of UAS data

High-throughput field phenotyping in cereals and implications in plant ecophysiology

[eng] Global climate change effects on agroecosystems together with increasing world population is already threatening food security and endangering ecosystem stability. Meet global food demand with crops production under climate change scenario is the core challenge in plant research nowadays. Thus, there is an urgent need to better understand the underpinning mechanisms of plant acclimation to stress conditions contributing to obtain resilient crops. Also, it is essential to develop new methods in plant research that permit to better characterize non-destructively plant traits of interest. In this sense, the advance in plant phenotyping research by high throughput systems is key to overcome these challenges, while its verification in the field may clear doubts on its feasibility. To this aim, this thesis focused on wheat and secondarily on maize as study species as they make up the major staple crops worldwide. A large panoply of phenotyping methods was employed in these works, ranging from RGB and hyperspectral sensing to metabolomic characterization, besides of other more conventional traits. All research was performed with trials grown in the field and diverse stressor conditions representative of major constrains for plant growth and production were studied: water stress, nitrogen deficiency and disease stress. Our results demonstrated the great potential of leave-to-canopy color traits captured by RGB sensors for in-field phenotyping, as they were accurate and robust indicators of grain yield in wheat and maize under disease and nitrogen deficiency conditions and of leaf nitrogen concentration in maize. On the other hand, the characterization of the metabolome of wheat tissues contributed to elucidate the metabolic mechanisms triggered by water stress and their relationship with high yielding performance, providing some potential biomarkers for higher yields and stress adaptation. Spectroscopic studies in wheat highlighted that leaf dorsoventrality may affect more than water stress on the reflected spectrum and consequently the performance of the multispectral/hyperspectral approaches to assess yield or any other relevant phenotypic trait. Anatomy, pigments and water changes were responsible of reflectance differences and the existence of leaf-side-specific responses were discussed. Finally, the use of spectroscopy for the estimation of the metabolite profiles of wheat organs showed promising for many metabolites which could pave the way for a new generation phenotyping. We concluded that future phenotyping may benefit from these findings in both the low-cost and straightforward methods and the more complex and frontier technologies.[cat] Els efectes del canvi climàtic sobre els agro-ecosistemes i l’increment de la població mundial posa en risc la seguretat alimentària i l’estabilitat dels ecosistemes. Actualment, satisfer les demandes de producció d’aliments sota l’escenari del canvi climàtic és el repte central a la Biologia Vegetal. Per això, és indispensable entendre els mecanismes subjacents de l’aclimatació a l’estrès que permeten obtenir cultius resilients. També és precís desenvolupar nou mètodes de recerca que permetin caracteritzar de manera no destructiva els trets d’interès. L’avenç del fenotipat vegetal amb sistemes d’alt rendiment és clau per abordar aquests reptes. La present tesi s’enfoca en el blat i secundàriament en el panís com a espècies d’estudi ja que constitueixen els cultius bàsics arreu del món. Un ampli ventall de mètodes de fenotipat s’han utilitzat, des sensors RGB a híper-espectrals fins a la caracterització metabolòmica. La recerca s’ha dut a terme en assajos de camp i s’han avaluat diversos tipus d’estrès representatius de les majors limitacions pel creixement i producció vegetal: estrès hídric i biòtic i deficiència de nitrogen. Els resultats demostraren el gran potencial dels trets del color RGB (des de la planta a la capçada) pel fenotipat de camp, ja que foren indicadors precisos del rendiment a blat i panís sota condicions de malaltia i deficiència de nitrogen i de la concentració de nitrogen foliar a panís. La caracterització metabolòmica de teixits de blat contribuí a esbrinar els processos metabòlics endegats per l’estrès hídric i la seva relació amb comportament genotípic, proporcionant bio-marcadors potencials per rendiments més alts i l’adaptació a l’estrès. Estudis espectroscòpics en blat van demostrar que la dorsoventralitat pot afectar més que l’estrès hídric sobre l’espectre de reflectància i consegüentment sobre el comportament de les aproximacions multi/híper-espectrals per avaluar el rendiment i d’altres trets fenotípics com anatòmics i contingut de pigments. Finalment, l’ús de l’espectroscòpia per l’estimació del contingut metabòlic als teixits de blat resulta prometedor per molts metabòlits, la qual cosa obre les portes per a un fenotipat de nova generació. El fenotipat pot beneficiar-se d’aquestes troballes, tant en els mètodes de baix cost com de les tecnologies més sofisticades i d’avantguarda

Estimating Photosynthetic Attributes from High-Throughput Canopy Hyperspectral Sensing in Sorghum

Sorghum, a genetically diverse C(4) cereal, is an ideal model to study natural variation in photosynthetic capacity. Specific leaf nitrogen (SLN) and leaf mass per leaf area (LMA), as well as, maximal rates of Rubisco carboxylation (V (cmax)), phosphoenolpyruvate (PEP) carboxylation (V (pmax)), and electron transport (J (max)), quantified using a C(4) photosynthesis model, were evaluated in two field-grown training sets (n = 169 plots including 124 genotypes) in 2019 and 2020. Partial least square regression (PLSR) was used to predict V (cmax) (R (2) = 0.83), V (pmax) (R (2) = 0.93), J (max) (R (2) = 0.76), SLN (R (2) = 0.82), and LMA (R (2) = 0.68) from tractor-based hyperspectral sensing. Further assessments of the capability of the PLSR models for V (cmax), V (pmax), J (max), SLN, and LMA were conducted by extrapolating these models to two trials of genome-wide association studies adjacent to the training sets in 2019 (n = 875 plots including 650 genotypes) and 2020 (n = 912 plots with 634 genotypes). The predicted traits showed medium to high heritability and genome-wide association studies using the predicted values identified four QTL for V (cmax) and two QTL for J (max). Candidate genes within 200 kb of the V (cmax) QTL were involved in nitrogen storage, which is closely associated with Rubisco, while not directly associated with Rubisco activity per se. J (max) QTL was enriched for candidate genes involved in electron transport. These outcomes suggest the methods here are of great promise to effectively screen large germplasm collections for enhanced photosynthetic capacity