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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.