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An acceleration of the genetic advances of durum wheat, as a major crop for the Mediterranean region, is required, but phenotyping still represents a bottleneck for breeding. This study aims to define durum wheat ideotypes under Mediterranean conditions by selecting the most suitable phenotypic remote sensing traits among different ones informing on characteristics related with leaf pigments/photosynthetic status, crop water status, and crop growth/green biomass. A set of 24 post-green revolution durum wheat cultivars were assessed in a wide set of 19 environments, accounted as the specific combinations of a range of latitudes in Spain, under different management conditions (water regimes and planting dates), through 3 consecutive years. Thus, red-green-blue and multispectral derived vegetation indices and canopy temperature were evaluated at anthesis and grain filling. The potential of the assessed remote sensing parameters alone and all combined as grain yield (GY) predictors was evaluated through random forest regression models performed for each environment and phenological stage. Biomass and plot greenness indicators consistently proved to be reliable GY predictors in all of the environments tested for both phenological stages. For the lowest-yielding environment, the contribution of water status measurements was higher during anthesis, whereas, for the highest-yielding environments, better predictions were reported during grain filling. Remote sensing traits measured during the grain filling and informing on pigment content and photosynthetic capacity were highlighted under the environments with warmer conditions, as the late-planting treatments. Overall, canopy greenness indicators were reported as the highest correlated traits for most of the environments and regardless of the phenological moment assessed. The addition of carbon isotope composition of mature kernels was attempted to increase the accuracies, but only a few were slightly benefited, as differen

Durum wheat is an important cereal that is widely grown in the Mediterranean basin. In addition to high yield, grain quality traits are of high importance for farmers. The strong influence of climatic conditions makes the improvement of grain quality traits, like protein content, vitreousness, and test weight, a challenging task. Evaluation of quality traits post-harvest is time- and labor-intensive and requires expensive equipment, such as near-infrared spectroscopes or hyperspectral imagers. Predicting not only yield but also important quality traits in the field before harvest is of high value for breeders aiming to optimize resource allocation. Implementation of efficient approaches for trait prediction, such as the use of high-resolution spectral data acquired by a multispectral camera mounted on unmanned aerial vehicles (UAVs), needs to be explored. In this study, we have acquired multispectral image data with an 11-band multispectral camera mounted on a UAV and analyzed the data with machine learning (ML) models to predict grain yield and important quality traits in breeding micro-plots. Combining 11-band multispectral data for 34 cultivars and 16 environments allowed to develop ML models with good prediction capability. Applying the trained models to test sets explained a considerable degree of phenotypic variance with good accuracy showing r squared values of 0.84, 0.69, 0.64, and 0.61 and normalized root mean squared errors of 0.17, 0.07, 0.14, and 0.03 for grain yield, protein content, vitreousness, and test weight, respectively.

Progress in high-throughput phenotyping and genomics provides the potential to understand the genetic basis of plant functional differentiation. We developed a semi-automatic methodology based on unmanned aerial vehicle (UAV) imagery for deriving tree-level phenotypes followed by genome-wide association study (GWAS). An RGB-based point cloud was used for tree crown identification in a common garden of Pinus halepensis in Spain. Crowns were combined with multispectral and thermal orthomosaics to retrieve growth traits, vegetation indices and canopy temperature. Thereafter, GWAS was performed to analyse the association between phenotypes and genomic variation at 235 single nucleotide polymorphisms (SNPs). Growth traits were associated with 12 SNPs involved in cellulose and carbohydrate metabolism. Indices related to transpiration and leaf water content were associated with six SNPs involved in stomata dynamics. Indices related to leaf pigments and leaf area were associated with 11 SNPs involved in signalling and peroxisome metabolism. About 16-20% of trait variance was explained by combinations of several SNPs, indicating polygenic control of morpho-physiological traits. Despite a limited availability of markers and individuals, this study is provides a successful proof-of-concept for the combination of high-throughput UAV-based phenotyping with cost-effective genotyping to disentangle the genetic architecture of phenotypic variation in a widespread conifer.

Knowledge of the agronomic and physiological traits associated with genetic gains in yield is essential to improve understanding of yield-limiting factors and to inform future breeding strategies. The aim of this paper is to dissect the agronomic and physiological traits related to genetic gain and to propose an ideotype with high yield that is best adapted to Spanish Mediterranean environments. Six semi-dwarf (i.e. modern) durum wheat genotypes were grown in a wide range of growing conditions in Spain during two successive years. Diverse agronomic, physiological and leaf morphological traits were evaluated. Kernels spike-1 was the yield component most affected by the genetic gain. While no interaction between genotype and growing conditions existed for grain yield, the more productive genotypes were characterized by a plant height of around 85 cm, small erect flag leaves, more open stomata, a better balance between N sources and N sinks and a higher capacity to re-fix CO2 respired by the grain. Moreover, in general the non-laminar parts of the plants play a key role in providing assimilates during grain filling. The high heritability of most of the studied parameters allows their consideration as traits for phenotyping durum wheat better adapted to a wide range of Mediterranean conditions.

RGB imagery has been widely used for crop management practices and phenotyping applications in recent years. Although RGB wavelengths (400-700 nm) are not able to capture all essential plant data (such as with full ultraviolet, near and long infrared wavelength coverage), RGB cameras are the most common types of cameras and are among the versatile imaging devices for proximal remote sensing applications. Deep learning strategies have improved a wide range of processes and deep learning concepts can be included in many applications. This work uses the Very Deep Super-Resolution (VDSP) technique to improve low-resolution RGB images in order to study grain yield assessment in wheat using vegetation indexes. The results show no significant differences between indexes calculated from low-resolution images and low-resolution images processed using VDSP with grain yield.

Although the relevance of spike bracts in stress acclimation and contribution to wheat yield was recently revealed, the metabolome of this organ and its response to water stress is still unknown. The metabolite profiles of flag leaves, glumes and lemmas were characterized under contrasting field water regimes in five durum wheat cultivars. Water conditions during growth were characterized through spectral vegetation indices, canopy temperature and isotope composition. Spike bracts exhibited better coordination of carbon and nitrogen metabolisms than the flag leaves in terms of photorespiration, nitrogen assimilation and respiration paths. This coordination facilitated an accumulation of organic and amino acids in spike bracts, especially under water stress. The metabolomic response to water stress also involved an accumulation of antioxidant and drought tolerance related sugars, particularly in the spikes. Furthermore, certain cell wall, respiratory and protective metabolites were associated with genotypic outperformance and yield stability. In addition, grain yield was strongly predicted by leaf and spike bracts metabolomes independently. This study supports the role of the spike as a key organ during wheat grain filling, particularly under stress conditions and provides relevant information to explore new ways to improve wheat productivity including potential biomarkers for yield prediction.