High-throughput crop phenotyping, particularly under field conditions, is nowadays perceived as a key factor limiting crop genetic advance. Phenotyping not only facilitates conventional breeding, but it is necessary to fully exploit the capabilities of molecular breeding, and it can be exploited to predict breeding targets for the years ahead at the regional level through more advanced simulation models and decision support systems. In terms of phenotyping, it is necessary to determined which selection traits are relevant in each situation, and which phenotyping tools/ methods are available to assess such traits. Remote sensing methodologies are currently the most popular approaches, even when lab-based analyses are still relevant in many circumstances. On top of that, data processing and automation, together with machine learning/deep learning are contributing to the wide range of applications for phenotyping. This review addresses spectral and red-green-blue sensing as the most popular remote sensing approaches, alongside stable isotope composition as an example of a lab-based tool, and root phenotyping, which represents one of the frontiers for field phenotyping. Further, we consider the two most promising forms of aerial platforms (unmanned aerial vehicle and satellites) and some of the emerging data-processing techniques. The review includes three Boxes that examine specific case studies.

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Several studies have highlighted the negative effects of ozone (O3) on wheat development and productivity. The negative effects of O3 are mediated by changes in photosynthetic carbon and nitrogen metabolism, which are difficult and time-consuming to assess and are thus only measured sporadically throughout the plant cycle. Stable isotope measurements in grains can help integrate the effects of chronic O3 exposure over the lifespan of the plant. This particular study focuses on the extent to which the stomatal conductance and productivity of Mediterranean wheat are related to carbon and nitrogen isotopic signatures under chronic O3 exposure. An open top chamber experiment was designed to analyse the effects of the pollutant on 12 Spanish wheat genotypes, which included modern cultivars, old cultivars and landraces. Four O3 treatments were considered. Stomatal conductance (gs) measurements were carried out during anthesis, and yield and nitrogen content parameters were taken at maturity, along with the carbon (δ13C) and nitrogen (δ15N) isotopic composition measured in grains. Modern and old cultivars responded similarly to O3 and were sensitive to the pollutant regarding yield parameters and gs, while landraces were more O3-tolerant. Grain δ13C had a strong negative correlation with grain yield and stomatal conductance across genotypes and O3 conditions, and increased under higher O3 concentrations, showing its capacity to integrate O3 stress throughout the wheat cycle. Meanwhile, a higher nitrogen concentration in grains, coupled with smaller grains, led to an overall decreased grain nitrogen yield under higher O3 concentrations. This nitrogen concentration effect within the grain differed among genotypes bred at different ages, following their respective O3-sensitivity. δ15N showed a possible indirect effect of O3 on nitrogen redistribution, particularly under the highest O3 concentration. The correlations of δ15N and δ13C to the usual effects of ozone on the plant suggest their potential as indicators of chronic ozone exposure.

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