Defining durum wheat ideotypes adapted to Mediterranean environments through remote sensing traits.

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 differences in water status among cultivars were already accounted by the measurement of canopy temperature.

Analysis of durum wheat photosynthetic organs during grain filling reveals the ear as a water stress-tolerant organ and the peduncle as the largest pool of primary metabolites.

MAIN CONCLUSION: The pool of carbon- and nitrogen-rich metabolites is quantitatively relevant in non-foliar photosynthetic organs during grain filling, which have a better response to water limitation than flag leaves. The response of durum wheat to contrasting water regimes has been extensively studied at leaf and agronomic level in previous studies, but the water stress effects on source-sink dynamics, particularly non-foliar photosynthetic organs, is more limited. Our study aims to investigate the response of different photosynthetic organs to water stress and to quantify the pool of carbon and nitrogen metabolites available for grain filling. Five durum wheat varieties were grown in field trials in the Spanish region of Castile and León under irrigated and rainfed conditions. Water stress led to a significant decrease in yield, biomass, and carbon and nitrogen assimilation, improved water use efficiency, and modified grain quality traits in the five varieties. The pool of carbon (glucose, glucose-6-phosphate, fructose, sucrose, starch, and malate) and nitrogen (glutamate, amino acids, proteins and chlorophylls) metabolites in leaf blades and sheaths, peduncles, awns, glumes and lemmas were also analysed. The results showed that the metabolism of the blades and peduncles was the most susceptible to water stress, while ear metabolism showed higher stability, particularly at mid-grain filling. Interestingly, the total metabolite content per organ highlighted that a large source of nutrients, which may be directly involved in grain filling, are found outside the blades, with the peduncles being quantitatively the most relevant. We conclude that yield improvements in our Mediterranean agro-ecosystem are highly linked to the success of shoots in producing ears and a higher number of grains, while grain filling is highly dependent on the capacity of non-foliar organs to fix CO2 and N. The ear organs show higher stress resilience than other organs, which deserves our attention in future breeding programmes.

Source-Sink Dynamics in Field-Grown Durum Wheat Under Contrasting Nitrogen Supplies: Key Role of Non-Foliar Organs During Grain Filling.

The integration of high-throughput phenotyping and metabolic approaches is a suitable strategy to study the genotype-by-environment interaction and identify novel traits for crop improvement from canopy to an organ level. Our aims were to study the phenotypic and metabolic traits that are related to grain yield and quality at canopy and organ levels, with a special focus on source-sink coordination under contrasting N supplies. Four modern durum wheat varieties with contrasting grain yield were grown in field conditions under two N fertilization levels in north-eastern Spain. We evaluated canopy vegetation indices taken throughout the growing season, physiological and metabolic traits in different photosynthetic organs (flag leaf blade, sheath, peduncle, awn, glume, and lemma) at anthesis and mid-grain filling stages, and agronomic and grain quality traits at harvest. Low N supply triggered an imbalance of C and N coordination at the whole plant level, leading to a reduction of grain yield and nutrient composition. The activities of key enzymes in C and N metabolism as well as the levels of photoassimilates showed that each organ plays an important role during grain filling, some with a higher photosynthetic capacity, others for nutrient storage for later stages of grain filling, or N assimilation and recycling. Interestingly, the enzyme activities and sucrose content of the ear organs were positively associated with grain yield and quality, suggesting, together with the regression models using isotope signatures, the potential contribution of these organs during grain filling. This study highlights the use of holistic approaches to the identification of novel targets to improve grain yield and quality in C3 cereals and the key role of non-foliar organs at late-growth stages.

Dataset of above and below ground traits assessed in Durum wheat cultivars grown under Mediterranean environments differing in water and temperature conditions.

Ideotypic characteristics of durum wheat associated with higher yield under different water and temperature regimes were studied under Mediterranean conditions. The focus of this paper is to provide raw and supplemental data from the research article entitled "Durum wheat ideotypes in Mediterranean environments differing in water and temperature conditions" [1], which aims to define specific durum wheat ideotypes according to their responses to different agronomic conditions. In this context, six modern (i.e. post green revolution) genotypes with contrasting yield performance (i.e. high vs low yield) were grown during two consecutive years under different treatments: (i) winter planting under support-irrigation conditions, (ii) winter planting under rainfed conditions, (iii) late planting under support-irrigation. Trials were conducted at the INIA station of Colmenar de Oreja (Madrid). Different traits were assessed to inform about water status (canopy temperature at anthesis and stable carbon isotope composition (δ13C) of the flag leaf and mature grains), root performance (root traits and the oxygen isotope composition (δ18O) in the stem base water), phenology (days from sowing to heading), nitrogen status/photosynthetic capacity (nitrogen content and stable isotope composition (δ15N) of the flag leaf and mature grain together with the pigment contents and the nitrogen balance index (NBI) of the flag leaf), crop growth (plant height (PH) and the normalized difference vegetation index (NDVI) at anthesis), grain yield and agronomic yield components. For most of the parameters assessed, data analysis demonstrated significant differences among genotypes within each treatment. The level of significance was determined using the Tukey-b test on independent samples, and ideotypes were modelled from the results of principle component analysis. The present data shed light on traits that help to define specific ideotype characteristics that confer genotypic adaptation to a wide range of agronomic conditions produced by variations in planting date, water conditions and season.

Preharvest phenotypic prediction of grain quality and yield of durum wheat using multispectral imaging.

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.

Agronomic and physiological traits related to the genetic advance of semi-dwarf durum wheat: The case of Spain.

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.

Metabolome Profiling Supports the Key Role of the Spike in Wheat Yield Performance.

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.

Cereal Crop Ear Counting in Field Conditions Using Zenithal RGB Images.

Ear density, or the number of ears per square meter (ears/m2), is a central focus in many cereal crop breeding programs, such as wheat and barley, representing an important agronomic yield component for estimating grain yield. Therefore, a quick, efficient, and standardized technique for assessing ear density would aid in improving agricultural management, providing improvements in preharvest yield predictions, or could even be used as a tool for crop breeding when it has been defined as a trait of importance. Not only are the current techniques for manual ear density assessments laborious and time-consuming, but they are also without any official standardized protocol, whether by linear meter, area quadrant, or an extrapolation based on plant ear density and plant counts postharvest. An automatic ear counting algorithm is presented in detail for estimating ear density with only sunlight illumination in field conditions based on zenithal (nadir) natural color (red, green, and blue [RGB]) digital images, allowing for high-throughput standardized measurements. Different field trials of durum wheat and barley distributed geographically across Spain during the 2014/2015 and 2015/2016 crop seasons in irrigated and rainfed trials were used to provide representative results. The three-phase protocol includes crop growth stage and field condition planning, image capture guidelines, and a computer algorithm of three steps: (i) a Laplacian frequency filter to remove low- and high-frequency artifacts, (ii) a median filter to reduce high noise, and (iii) segmentation and counting using local maxima peaks for the final count. Minor adjustments to the algorithm code must be made corresponding to the camera resolution, focal length, and distance between the camera and the crop canopy. The results demonstrate a high success rate (higher than 90%) and R2 values (of 0.62-0.75) between the algorithm counts and the manual image-based ear counts for both durum wheat and barley.

Leaf dorsoventrality as a paramount factor determining spectral performance in field-grown wheat under contrasting water regimes.

The effects of leaf dorsoventrality and its interaction with environmentally induced changes in the leaf spectral response are still poorly understood, particularly for isobilateral leaves. We investigated the spectral performance of 24 genotypes of field-grown durum wheat at two locations under both rainfed and irrigated conditions. Flag leaf reflectance spectra in the VIS-NIR-SWIR (visible-near-infrared-short-wave infrared) regions were recorded in the adaxial and abaxial leaf sides and at the canopy level, while traits providing information on water status and grain yield were evaluated. Moreover, leaf anatomical parameters were measured in a subset of five genotypes. The spectral traits studied were more affected by the leaf side than by the water regime. Leaf dorsoventral differences suggested higher accessory pigment content in the abaxial leaf side, while water regime differences were related to increased chlorophyll, nitrogen, and water contents in the leaves in the irrigated treatment. These variations were associated with anatomical changes. Additionally, leaf dorsoventral differences were less in the rainfed treatment, suggesting the existence of leaf-side-specific responses at the anatomical and biochemical level. Finally, the accuracy in yield prediction was enhanced when abaxial leaf spectra were employed. We concluded that the importance of dorsoventrality in spectral traits is paramount, even in isobilateral leaves.

Wheat ear counting in-field conditions: high throughput and low-cost approach using RGB images.

BACKGROUND: The number of ears per unit ground area (ear density) is one of the main agronomic yield components in determining grain yield in wheat. A fast evaluation of this attribute may contribute to monitoring the efficiency of crop management practices, to an early prediction of grain yield or as a phenotyping trait in breeding programs. Currently the number of ears is counted manually, which is time consuming. Moreover, there is no single standardized protocol for counting the ears. An automatic ear-counting algorithm is proposed to estimate ear density under field conditions based on zenithal color digital images taken from above the crop in natural light conditions. Field trials were carried out at two sites in Spain during the 2014/2015 crop season on a set of 24 varieties of durum wheat with two growing conditions per site. The algorithm for counting uses three steps: (1) a Laplacian frequency filter chosen to remove low and high frequency elements appearing in an image, (2) a Median filter to reduce high noise still present around the ears and (3) segmentation using Find Maxima to segment local peaks and determine the ear count within the image. RESULTS: The results demonstrate high success rate (higher than 90%) between the algorithm counts and the manual (image-based) ear counts, and precision, with a low standard deviation (around 5%). The relationships between algorithm ear counts and grain yield was also significant and greater than the correlation with manual (field-based) ear counts. In this approach, results demonstrate that automatic ear counting performed on data captured around anthesis correlated better with grain yield than with images captured at later stages when the low performance of ear counting at late grain filling stages was associated with the loss of contrast between canopy and ears. CONCLUSIONS: Developing robust, low-cost and efficient field methods to assess wheat ear density, as a major agronomic component of yield, is highly relevant for phenotyping efforts towards increases in grain yield. Although the phenological stage of measurements is important, the robust image analysis algorithm presented here appears to be amenable from aerial or other automated platforms.

The Plant-Transpiration Response to Vapor Pressure Deficit (VPD) in Durum Wheat Is Associated With Differential Yield Performance and Specific Expression of Genes Involved in Primary Metabolism and Water Transport.

The regulation of plant transpiration was proposed as a key factor affecting transpiration efficiency and agronomical adaptation of wheat to water-limited Mediterranean environments. However, to date no studies have related this trait to crop performance in the field. In this study, the transpiration response to increasing vapor pressure deficit (VPD) of modern Spanish semi-dwarf durum wheat lines was evaluated under controlled conditions at vegetative stage, and the agronomical performance of the same set of lines was assessed at grain filling as well as grain yield at maturity, in Mediterranean environments ranging from water stressed to good agronomical conditions. A group of linear-transpiration response (LTR) lines exhibited better performance in grain yield and biomass compared to segmented-transpiration response (STR) lines, particularly in the wetter environments, whereas the reverse occurred only in the most stressed trial. LTR lines generally exhibited better water status (stomatal conductance) and larger green biomass (vegetation indices) during the reproductive stage than STR lines. In both groups, the responses to growing conditions were associated with the expression levels of dehydration-responsive transcription factors (DREB) leading to different performances of primary metabolism-related enzymes. Thus, the response of LTR lines under fair to good conditions was associated with higher transcription levels of genes involved in nitrogen (GS1 and GOGAT) and carbon (RCBL) metabolism, as well as water transport (TIP1.1). In conclusion, modern durum wheat lines differed in their response to water loss, the linear transpiration seemed to favor uptake and transport of water and nutrients, and photosynthetic metabolism led to higher grain yield except for very harsh drought conditions. The transpiration response to VPD may be a trait to further explore when selecting adaptation to specific water conditions.

Contribution of the ear and the flag leaf to grain filling in durum wheat inferred from the carbon isotope signature: genotypic and growing conditions effects.

The ear, together with the flag leaf, is believed to play a major role as a source of assimilates during grain filling in C3 cereals. However, the intrusive nature of most of the available methodologies prevents reaching conclusive results in this regard. This study compares the carbon isotope composition (δ(13)C) in its natural abundance in the water-soluble fractions of the flag leaf blade and the ear with the δ(13)C of mature kernels to assess the relative contribution of both organs to grain filling in durum wheat (Triticum turgidum L. var. durum). The relative contribution of the ear was higher in landraces compared to modern cultivars, as well as in response to nitrogen fertilization and water stress. Such genotypic and environmentally driven differences were associated with changes in harvest index (HI), with the relative contribution of the ear being negatively associated with HI. In the case of the genotypic differences, the lower relative contribution of the ear in modern cultivars compared with landraces is probably associated with the appearance in the former of a certain amount of source limitation driven by a higher HI. In fact, the relative contribution of the ear was far more responsive to changes in HI in modern cultivars compared with landraces.