Non-foliar photosynthesis and nitrogen assimilation influence grain yield in durum wheat regardless of water conditions.

There is a need to generate improved crop varieties adapted to the ongoing changes in the climate. We studied durum wheat canopy and central metabolism of six different photosynthetic organs in two yield-contrasting varieties. The aim was to understand the mechanisms associated with the water stress response and yield performance. Water stress strongly reduced grain yield, plant biomass, and leaf photosynthesis, and down-regulated C/N-metabolism genes and key protein levels, which occurred mainly in leaf blades. By contrast, higher yield was associated with high ear dry weight and lower biomass and ears per area, highlighting the advantage of reduced tillering and the consequent improvement in sink strength, which promoted C/N metabolism at the whole plant level. An improved C metabolism in blades and ear bracts and N assimilation in all photosynthetic organs facilitated C/N remobilization to the grain and promoted yield. Therefore, we propose that further yield gains in Mediterranean conditions could be achieved by considering the source-sink dynamics and the contribution of non-foliar organs, and particularly N assimilation and remobilization during the late growth stages. We highlight the power of linking phenotyping with plant metabolism to identify novel traits at the whole plant level to support breeding programmes.

Crop phenotyping in a context of global change: What to measure and how to do it.

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.

Gene expression and physiological responses to salinity and water stress of contrasting durum wheat genotypes.

Elucidating the relationships between gene expression and the physiological mechanisms remains a bottleneck in breeding for resistance to salinity and drought. This study related the expression of key target genes with the physiological performance of durum wheat under different combinations of salinity and irrigation. The candidate genes assayed included two encoding for the DREB (dehydration responsive element binding) transcription factors TaDREB1A and TaDREB2B, another two for the cytosolic and plastidic glutamine synthetase (TaGS1 and TaGS2), and one for the specific Na(+) /H(+) vacuolar antiporter (TaNHX1). Expression of these genes was related to growth and different trait indicators of nitrogen metabolism (nitrogen content, stable nitrogen isotope composition, and glutamine synthetase and nitrate reductase activities), photosynthetic carbon metabolism (stable carbon isotope composition and different gas exchange traits) and ion accumulation. Significant interaction between genotype and growing conditions occurred for growth, nitrogen content, and the expression of most genes. In general terms, higher expression of TaGS1, TaGS2, TaDREB2B, and to a lesser extent of TaNHX1 were associated with a better genotypic performance in growth, nitrogen, and carbon photosynthetic metabolism under salinity and water stress. However, TaDREB1A was increased in expression under stress compared with control conditions, with tolerant genotypes exhibiting lower expression than susceptible ones.

Wheat ear carbon assimilation and nitrogen remobilization contribute significantly to grain yield.

The role of wheat ears as a source of nitrogen (N) and carbon (C) in the grain filling process has barely been studied. To resolve this question, five wheat genotypes were labeled with 15 N-enriched nutrient solution. N remobilization and absorption were estimated via the nitrogen isotope composition of total organic matter and Rubisco. Gas exchange analyses showed that ear photosynthesis contributed substantially to grain filling in spite of the great loss of C due to respiration. Of the total kernel N, 64.7% was derived from the N acquired between sowing and anthesis, while the remaining 35.3% was derived from the N acquired between anthesis and maturity. In addition, 1.87 times more N was remobilized to the developing kernel from the ear than from the flag leaf. The higher yielding genotypes showed an increased N remobilization to the kernel compared to the lower yielding genotypes. In addition, the higher yielding genotypes remobilized more N from the ears to the kernel than the lower yielding genotypes, while the lower yielding genotypes remobilized more N from the flag leaf to the kernel. Therefore, the ears contribute significantly toward fulfilling C and N demands during grain filling.

Quinoa panicles contribute to carbon assimilation and are more tolerant to salt stress than leaves.

Contribution of inflorescences to seed filling have attracted great attention given the resilience of this photosynthetic organ to stressful conditions. However, studies have been almost exclusively focused to small grain cereals. In this study, we aimed to explore these responses in quinoa, as a climate resilient seed crop of elevated economic and nutritious potential. We compared the physiological and metabolic performance of panicles and leaves of two quinoa cultivars growing under contrasting salinity levels. Plant growth, photosynthetic and transpiratory gas exchange and chlorophyll fluorescence were monitored in inflorescences and leaves throughout the experiment. At flowering stage, young and mature leaves and panicles were sampled for key metabolic markers related to carbon, nitrogen and secondary metabolisms. When subjected to salt stress, panicles showed attenuated declines on photosynthesis, water use, pigments, amino acids, and protein levels as compared to leaves. In fact, the assimilation rates, together with a high hexose content evidenced an active photosynthetic role of the panicle under optimal and salt stress conditions. Moreover, we also found significant genotypic variability for physiological and metabolic traits of panicles and leaves, which emphasizes the study of genotype-dependent stress responses at the whole plant level. We conclude that quinoa panicles are less affected by salt stress than leaves, which encourages further research and exploitation of this organ for crop improvement and stress resilience considering the high natural diversity.

Comparative response of δ13C, δ18O and δ15N in durum wheat exposed to salinity at the vegetative and reproductive stages.

This study compared the performance of the stable isotope composition of carbon (δ(13) C), oxygen (δ(18) O) and nitrogen (δ(15) N) by tracking plant response and genotypic variability of durum wheat to different salinity conditions. To that end, δ(13) C, δ(18) O and δ(15) N were analysed in dry matter (dm) and the water-soluble fraction (wsf) of leaves from plants exposed to salinity, either soon after plant emergence or at anthesis. The δ(13) C and δ(18) O of the wsf recorded the recent growing conditions, including changes in evaporative conditions. Regardless of the plant part (dm or wsf), δ(13) C and δ(18) O increased and δ(15) N decreased in response to stress. When the stress conditions were established just after emergence, δ(15) N and δ(13) C correlated positively with genotypic differences in biomass, whereas δ(18) O correlated negatively in the most severe treatment. When the stress conditions were imposed at anthesis, relationships between the three isotope signatures and biomass were only significant and positive within the most severe treatments. The results show that nitrogen metabolism, together with stomatal limitation, is involved in the genotypic response to salinity, with the relative importance of each factor depending on the severity and duration of the stress as well as the phenological stage that the stress occurs.

Combined use of δ¹³C, δ18O and δ15N tracks nitrogen metabolism and genotypic adaptation of durum wheat to salinity and water deficit.

• Accurate phenotyping remains a bottleneck in breeding for salinity and drought resistance. Here the combined use of stable isotope compositions of carbon (δ¹³C), oxygen (δ¹8O) and nitrogen (δ¹5N) in dry matter is aimed at assessing genotypic responses of durum wheat under different combinations of these stresses. • Two tolerant and two susceptible genotypes to salinity were grown under five combinations of salinity and irrigation regimes. Plant biomass, δ¹³C, δ¹8O and δ¹5N, gas-exchange parameters, ion and N concentrations, and nitrate reductase (NR) and glutamine synthetase (GS) activities were measured. • Stresses significantly affected all traits studied. However, only δ¹³C, δ¹8O, δ¹5N, GS and NR activities, and N concentration allowed for clear differentiation between tolerant and susceptible genotypes. Further, a conceptual model explaining differences in biomass based on such traits was developed for each growing condition. • Differences in acclimation responses among durum wheat genotypes under different stress treatments were associated with δ¹³C. However, except for the most severe stress, δ¹³C did not have a direct (negative) relationship to biomass, being mediated through factors affecting δ¹8O or N metabolism. Based upon these results, the key role of N metabolism in durum wheat adaptation to salinity and water stress is highlighted.

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.

Comparative Performances of Beneficial Microorganisms on the Induction of Durum Wheat Tolerance to Fusarium Head Blight.

Durum wheat production is seriously threatened by Fusarium head blight (FHB) attacks in Tunisia, and the seed coating by bio-agents is a great alternative for chemical disease control. This study focuses on evaluating, under field conditions, the effect of seed coating with Trichoderma harzianum, Meyerozyma guilliermondii and their combination on (i) FHB severity, durum wheat grain yield and TKW in three crop seasons, and (ii) on physiological parameters and the carbon and nitrogen content and isotope composition in leaves and grains of durum wheat. The results indicated that the treatments were effective in reducing FHB severity by 30 to 70% and increasing grain yield with an increased rate ranging from 25 to 68%, compared to the inoculated control. The impact of treatments on grain yield improvement was associated with higher NDVI and chlorophyll content and lower canopy temperature. Furthermore, the treatments mitigated the FHB adverse effects on N and C metabolism by resulting in a higher δ13Cgrain (13C/12Cgrain) and δ15Ngrain (15N/14Ngrain). Overall, the combination outperformed the other seed treatments by producing the highest grain yield and TKW. The high potency of seed coating with the combination suggests that the two microorganisms have synergetic or complementary impacts on wheat.

Exploring the Potential of Meyerozyma guilliermondii on Physiological Performances and Defense Response against Fusarium Crown Rot on Durum Wheat.

Coating seeds with bio-control agents is a potentially effective approach to reduce the usage of pesticides and fertilizers applied and protect the natural environment. This study evaluated the effect of seed coating with Meyerozyma guilliermondii, strain INAT (MT731365), on seed germination, plant growth and photosynthesis, and plant resistance against Fusarium culmorum, in durum wheat under controlled conditions. Compared to control plants, seed coating with M. guilliermondii promoted the wheat growth (shoot and roots length and biomass), and photosynthesis and transpiration traits (chlorophyll, ɸPSII, rates of photosynthesis and transpiration, etc.) together with higher nitrogen balance index (NBI) and lower flavonols and anthocyanins. At 21 days post infection with Fusarium, M. guilliermondii was found to reduce the disease incidence and the severity, with reduction rates reaching up to 31.2% and 30.4%, respectively, as well as to alleviate the disease damaging impact on photosynthesis and plant growth. This was associated with lower ABA, flavonols and anthocyanins, compared to infected control. A pivotal function of M. guilliermondii as an antagonist of F. culmorum and a growth promoter is discussed.

Effect of salinity and water stress during the reproductive stage on growth, ion concentrations, Delta 13C, and delta 15N of durum wheat and related amphiploids.

The physiological performance of durum wheat and two related amphiploids was studied during the reproductive stage under different combinations of salinity and irrigation. One triticale, one tritordeum, and four durum wheat genotypes were grown in pots in the absence of stress until heading, when six different treatments were imposed progressively. Treatments resulted from the combination of two irrigation regimes (100% and 35% of container water capacity) with three levels of water salinity (1.8, 12, and 17 dS m(-1)), and were maintained for nearly 3 weeks. Gas exchange and chlorophyll fluorescence and content were measured prior to harvest; afterwards shoot biomass and height were recorded, and Delta(13)C, delta(15)N, and the concentration of nitrogen (N), phosphorus, and several ions (K(+), Na(+), Ca(2+), Mg(2+)) were analysed in shoot material. Compared with control conditions (full irrigation with Hoagland normal) all other treatments inhibited photosynthesis through stomatal closure, accelerated senescence, and decreased biomass. Full irrigation with 12 dS m(-1) outperformed other stress treatments in terms of biomass production and physiological performance. Biomass correlated positively with N and delta(15)N, and negatively with Na(+) across genotypes and fully irrigated treatments, while relationships across deficit irrigation conditions were weaker or absent. Delta(13)C did not correlate with biomass across treatments, but it was the best trait correlating with phenotypic differences in biomass within treatments. Tritordeum produced more biomass than durum wheat in all treatments. Its low Delta(13)C and high K(+)/Na(+) ratio, together with a high potential growth, may underlie this finding. Mechanisms relating delta(15)N and Delta(13)C to biomass are discussed.

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.

A Novel Aspect of Essential Oils: Coating Seeds with Thyme Essential Oil induces Drought Resistance in Wheat.

Coating seeds with biostimulants is among the promising approaches in crop production to increase crop tolerance to drought stress. In this study, we evaluated the potential of coating durum wheat seeds of the cultivar 'Karim' with thyme essential oil on enhancing seed germination and seedling growth, and on plant growth promotion and induction of drought resistance. Coated seeds were pre-germinated, grown in hydroponics, and grown in pots under controlled well-watered and progressive water/nutrient stress conditions. Seed coating with thyme oil increased germination rate and enhanced seedling growth development in hydroponics. In the pot experiment, thyme oil increased, when well watered, root and shoot development, chlorophyll, nitrogen balance index (NBI), abscisic acid (ABA), anthocyanins and flavonoids in leaves, decreased nitrogen isotope composition (δ15N) and increased carbon isotope composition (δ13C) of shoots. Increasing water/nutrient stress in control plants induced higher accumulation of ABA and anthocyanins coupled with a transient decrease in chlorophyll and NBI, a decrease in shoot and root development, the Normalized Difference Vegetation Index (NDVI), shoot C content, δ15N, and an increase in δ13C, revealing the avoidance strategy adopted by the cultivar. Thyme oil had the potential to enhance the avoidance strategy by inducing roots elongation, reducing the loss of shoot and roots dry matter and chlorophyll, maintaining balanced NBI, an decreasing anthocyanins, flavonoids, and δ13C via maintaining lower ABA-mediated-stomatal closure. Thyme oil increased shoot N content and δ15N indicating preferential uptake of the 15N enriched NH4+. Coating seeds with thyme oil is suggested as a promising alternative approach to improve plant's water and nutrient status and to enhance drought resistance.

Identification of traits associated with barley yield performance using contrasting nitrogen fertilizations and genotypes.

Much attention has been paid to understanding the traits associated with crop performance and the associated underlying physiological mechanisms, with less effort done towards combining different plant scales, levels of observation, or including hybrids of autogamous species. We aim to identify mechanisms at canopy, leaf and transcript levels contributing to crop performance under contrasting nitrogen supplies in three barley genotypes, two hybrids and one commercial line. High nitrogen fertilization did not affect photosynthetic capacity on a leaf area basis and lowered nitrogen partial factor productivity past a certain point, but increased leaf area and biomass accumulation, parameters that were closely tracked using various different high throughput remote sensing based phenotyping techniques. These aspects, together with a larger catabolism of leaf nitrogen compounds amenable to sink translocation, contributed to higher crop production. Better crop yield and growth in hybrids compared to the line was linked to a nitrogen-saving strategy in source leaves to the detriment of larger sink size, as indicated by the lower leaf nitrogen content and downregulation of nitrogen metabolism and aquaporin genes. While these changes did not reduce photosynthesis capacity on an area basis, they were related with better nitrogen use in the hybrids compared with the line.

Relative Contribution of Nitrogen Absorption, Remobilization, and Partitioning to the Ear During Grain Filling in Chinese Winter Wheat.

Knowledge of the function of the ear as a key organ in the uptake, remobilization and partitioning of nitrogen is essential for understanding its contribution to grain filling and thus guiding future breeding strategies. In this work, four Chinese winter wheat genotypes were grown on a 15N-enriched nutrient solution. N absorption and further remobilization to the flag leaf, the ear and the mature grains were calculated via the 15N atom % excess. The results indicated that the high yields of the Chinese wheat genotype were determined by higher grain numbers per ear, with greater plant height and a larger ear size, while the thousand-grain weight did not affect grain yield. In the mature grains, 66.7% of total N was remobilized from the pre-anthesis accumulation in the biomass, while the remaining 33.3% was derived from the N taken up during post-anthesis. From anthesis to 2 weeks after the anthesis stage, the flag leaf remobilized 3.67 mg of N outwards and the ear remobilized 3.87 mg of N inwards from the pre-anthesis accumulation in each plant. The positive correlation between ear Nrem and grain Nrem indicated that the ear was an important organ for providing N to the grain, whereas the remobilized N stream from the leaves was not correlated with grain Nrem, thus indicating that flag leaf N was not translocated directly to the grain. The grain Nrem was negatively correlated with the ear N concentration throughout grain filling, which suggested that higher-yielding genotypes had better sink activity in the ear, while Rubisco played a critical role in N deposition. Therefore, to improve yield potential in wheat, the N accumulation in the ear and the subsequent remobilization of that stored N to the grains should be considered. N accumulation and remobilization in the ear may at least be valuable for Chinese breeding programs that aim at optimizing the sink/source balance to improve grain filling.

Molecular and physiological mechanisms associated with root exposure to mercury in barley.

Mercury (Hg) is a toxic metal that affects plant growth. Here the effect of Hg exposure on plant growth and leaf gas-exchange together with gene expression in roots is reported for barley. Hg was mainly accumulated in roots and only very small amounts were found in the shoots. Chlorophyll fluorescence and net photosynthesis were not affected by Hg. Nevertheless exposure to Hg reduced shoot and root growth, the shoot to root ratio, stomatal conductance, carbon isotope discrimination and expression of an aquaporin transcript, whereas abscisic acid related transcripts were over-expressed. These results suggested some degree of limitation to water uptake causing a moderate water stress when plants are exposed to Hg. Microarray (MapMan) analysis revealed changes in the transcription of genes involved in nitrogen metabolism, which were accompanied by decreased nitrogen concentrations in the shoots, together with an increase in transcripts associated with secondary metabolism, stress, inhibition of DNA synthesis/chromatin structure and cell organization elements. Moreover, Hg induced the expression of many transcripts known to be involved in the uptake, accumulation, transport and general responses to other heavy metals. It is concluded that barley is able to accumulate high amounts of Hg in roots through several transcriptional, metabolic and physiological adjustments.

Root traits and δ13C and δ18O of durum wheat under different water regimes.

Plant growth, root characteristics and the stable carbon (δ13C) and oxygen (δ18O) composition were studied in durum wheat. Four recombinant inbred lines with good agronomic adaptation were grown under well watered (WW) and water stress (WS) conditions until mid-grain filling in lysimeters. Gas exchange was measured in the flag leaf just before harvest and then the aerial dry matter (Aerial DM), root weight density (RWD) and root length density (RLD) and the specific root length (SRL) were evaluated and the δ13C and δ18O of the roots, the flag leaf blade and the spike were analysed. Water stress decreased stomatal conductance, plant accumulated transpiration and Aerial DM, whereas δ13C and δ18O increased. Genotypic differences were found for all gas-exchange and root traits and isotope signatures. Aerial DM was positively correlated with RLD, regardless of the water regime, whereas it was negatively correlated with δ13C and δ18O, but only under WW conditions. Moreover, RWD and RLD were negatively related to both δ13C and δ18O under the WW regime, but no clear pattern existed under WS. Our study supports the use of δ13C and δ18O as proxies for selecting root traits associated with higher growth in the absence of water stress.

Shoot δ15N gives a better indication than ion concentration or Δ13C of genotypic differences in the response of durum wheat to salinity.

We compared the performance of different physiological traits that reveal genotypic variations in tolerance to salinity in durum wheat. A set of 114 genotypes was grown in hydroponics for over 3 months. Three conditions: control, moderate (12 dS m-1) and severe (17 dS m-1) salinity, were maintained for nearly 8 weeks before harvest. The genotype biomass in control conditions correlated with the biomass at the two salinity levels. Subsequently, two subsets of 10 genotypes each were selected on the basis of extreme differences in biomass at the two salinity levels while showing relatively similar biomass in control conditions. Carbon isotope discrimination (Δ13C), nitrogen isotope composition (δ15N), and the concentration of nitrogen, phosphorus and several ions (K+, Na+, Ca2+, Mg2+) were analysed in the two subsets for the three treatments. At 12 dS m-1, K+ concentration, K+/Na+ ratio, Δ13C and δ15N correlated positively and Na+ correlated negatively with shoot biomass. Under control conditions and at 17 dS m-1 no correlation was observed. However, the trait that correlated best with genotypic differences in biomass was δ15N at 12 dS m-1. This trait was the first variable chosen at each of the two salinity levels in a stepwise analysis. We consider the possible mechanisms relating δ15N to biomass and the use of this isotopic signature as a selection trait.