Ús d'isòtops estables d'O, H, C com eines de selecció de rendiment potencial i adaptació a la sequera i deficiència de nitrogen en cereals C3 i C4

[cat] A la conca Mediterrània els principals factors que afecten el rendiment dels cultius agrícoles són la sequera i el dèficit de nitrogen. És necessària la recerca de noves eines de fenotipejat per accelerar la millora genètica pel rendiment potencial i l'adaptació dels cultius a condicions limitants. Aquesta Tesi ha estudiat composició isotòpica de carboni (δ13C), oxigen (δ18O) i hidrogen (δ2H), en la seva abundància natural, de diferents cereals. Referent a la δ13C només la composició dels grans madurs van mostrar correlacions fenotípiques (negatives) consistents amb el rendiment del gra (GY) en blat dur, sobretot sota condicions d'estrès moderat (Capítol 5). Aquesta Tesi també proposa l’ús de la δ13C com una aproximació per quantificar la contribució relativa dels diferents òrgans de la planta a l'ompliment del gra (Capítols 1 i 2). L’objectiu es poder emprar la δ13C per seleccionar cereals amb una major fotosíntesi de l'espiga. En blat dur l'aproximació de la δ13C va assignar un paper més gran a l'espiga (tant de l'òrgan sencer com de les arestes) en comparació a la fulla bandera i al peduncle (que representa els assimilats que provenen de les parts inferiors de l'espiga), especialment en les varietats antigues, amb independència de les condicions de creixement. Sota bones condicions agronòmiques, la contribució de les arestes de l’espiga a l'ompliment del gra de blat fariner també va ser més important que la de la fulla bandera. Finalment aquesta aproximació basada en la δ13C de diferents parts de planta es va comparar amb altres aproximacions convencionals com són l'ombrejat o l'aplicació de l'herbicida DCMU a la planta (Capítol 3). Els tractaments d'ombrejat de l'espiga i de la tija van contribuir de forma similar a l'ompliment del gra. Per contra, l'aproximació del DCMU va assignar un paper major a la fotosíntesi de la tija, però l'aplicació de l’herbicida a la tija també va afectar l'espiga, esbiaixant el pes final dels grans. Aquesta Tesi també va estudiar l'isòtop d'oxigen tant en blat de moro (Capítol 4) com en blat dur (Capítol 5). En el blat de moro, les correlacions genotípiques entre la δ18O i el GY van ser marginals, tot i que les sedes va ser l'òrgan que millor va correlacionar amb el GY. En el blat dur les correlacions fenotípiques entre la δ18O dels grans i el GY també van ser marginals. Només va correlacionar fortament la δ18O de l'aigua de la fulla quan es van combinar els dos règims hídrics (reg suplementari i sequera). L'absència de correlacions de la δ18O podria ajudar a descartar els teixits de les plantes que són més susceptibles als processos de fraccionament post-fotosintètic. Escollir l’òrgan idoni pot ajudar a millorar l’ús de la δ18O com a eina de millora de cultius. Per últim es va comparar la δ13C i la δ18O amb la δ2H (Capítol 5). La δ2H en els grans de blat dur va correlacionar contra el GY millor que els altres dos isòtops en condicions de sequera però combinant nivells de nitrogen contrastats. També es van observar correlacions genotípiques entre la δ2H dels foto-assimilats de l'espiga contra el GY. A més, la δ2H de les fulles va correlacionar més amb la δ13C que no pas amb la δ18O, el que suggereix que la δ2H dels foto-assimilats de la fulla no només està afectada per la transpiració i la conductància estomàtica sinó també per les reaccions fotosintètiques. A més, els valors baixos de la δ2H a l’espiga comparats amb els dels grans donen suport al paper fotosintètic de l’espiga, el que recolza els resultats obtinguts en els Capítols 1, 2 i 3.[eng] This Thesis has studied the isotope composition on its natural abundance of carbon (δ13C), oxygen (δ18O) and hydrogen (δ2H) as phenotypic traits for cereal breeding and crop adaptation to optimal and limited agronomic conditions. Regarding the δ13C, only mature grains showed consistent phenotypic correlations (negative) against grain yield in durum wheat, especially under moderate stress conditions. In addition δ13C is also proposed as a tool to quantify the relative contribution of different plant organs to grain filling. In durum wheat and bread wheat δ13C approach assigned a higher role to the ear (both whole body and awns) compared to the flag leaf and peduncle (which integrates the assimilates produced by photosynthetic organs below the ear), regardless of growing conditions. Finally, δ13C approach based on the different plant parts was compared with other conventional approaches, such as shading or herbicide DCMU application, which assigned on average a comparable contribution to the ear than the culm. This thesis also studied the δ18O in maize and durum wheat. In both crops, phenotypic correlations between δ18O and grain yield were marginal. Only δ18O of leaf water in durum wheat was strongly correlated with GY when combining two water regimes. The absence of such correlations will eventually help to understand the use of δ18O as a genotype selection tool for the adaptation of maize and other crops to drought. Finally the δ13C and δ18O were compared with δ2H in durum wheat. δ2H performed better than the other two isotopes predicting grain yield and nitrogen content under water stress but contrasting nitrogen regimes. Besides, genotypic correlations between δ2H in the ear water-soluble fraction and grain yield were observed. In addition, δ2H in the water soluble fraction of leaves was better correlated against δ13C than with δ18O, suggesting that δ2H of leaf photo-assimilated is affected not only by transpiration and stomatal conductance but also by the photosynthetic reactions. In addition, the low values observed in the δ2H in the ear compared to mature grains supported the photosynthetic role of the ear, which reinforced results obtained in other chapter of this Thesis

The nitrogen contribution of different plant parts to wheat grains: exploring genotype, water, and nitrogen effects

The flag leaf has been traditionally considered as the main contributor to grain nitrogen. However, during the reproductive stage, other organs besides the flag leaf may supply nitrogen to developing grains. Therefore, the contribution of the ear and other organs to the nitrogen supplied to the growing grains remains unclear. It is important to develop phenotypic tools to assess the relative contribution of different plant parts to the N accumulated in the grains of wheat which may helps to develop genotypes that use N more efficiently. We studied the effect of growing conditions (different levels of water and nitrogen in the field) on the nitrogen contribution of the spike and different vegetative organs of the plant to the grains. The natural abundance of 15 δ N and total N content in the flag blade, peduncle, whole spike, glumes and awns were compared to the 15 δ N and total N in mature grains to trace the origin of nitrogen redistribution to the grains. The 15 δ N and total N content of the different plant parts correlated positively with the 15 δ N and total N content of mature grains suggesting that all organs may contribute a portion of their N content to the grains. The potential contribution of the flag blade to grain N increased (by 46%) as the growing conditions improved, whereas the potential contribution of the glumes plus awns and the peduncle increased (46 and 31%, respectively) as water and nitrogen stress increased. In general, potential contribution of the ear providing N to growing grains was similar (42%) than that of the vegetative parts of the plants (30-40%), regardless of the growing conditions. Thus, the potential ear N content could be a positive trait for plant phenotyping, especially under water and nitrogen limiting conditions. In that sense, genotypic variability existed at least between old (tall) and modern (semidwarf) cultivars, with the ear from modern genotypes exhibiting less relative contribution to the total grain N. The combined use of 15 δ N and N content may be used as an affordable tool to assess the relative contribution of different plant parts to the grain N in wheat

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

Differential Flag Leaf and Ear Photosynthetic Performance Under Elevated (CO2) Conditions During Grain Filling Period in Durum Wheat

Elevated concentrations of CO2 (CO2) in plants with C3 photosynthesis metabolism, such as wheat, stimulate photosynthetic rates. However, photosynthesis tends to decrease as a function of exposure to high (CO2) due to down-regulation of the photosynthetic machinery, and this phenomenon is defined as photosynthetic acclimation. Considerable efforts are currently done to determine the effect of photosynthetic tissues, such us spike, in grain filling. There is good evidence that the contribution of ears to grain filling may be important not only under good agronomic conditions but also under high (CO2). The main objective of this study was to compare photoassimilate production and energy metabolism between flag leaves and glumes as part of ears of wheat (Triticum turgidum L. subsp. durum cv. Amilcar) plants exposed to ambient [a(CO2)] and elevated [e(CO2)] (CO2) (400 and 700 μmol mol–1, respectively). Elevated CO2 had a differential effect on the responses of flag leaves and ears. The ears showed higher gross photosynthesis and respiration rates compared to the flag leaves. The higher ear carbohydrate content and respiration rates contribute to increase the grain dry mass. Our results support the concept that acclimation of photosynthesis to e(CO2) is driven by sugar accumulation, reduction in N concentrations and repression of genes related to photosynthesis, glycolysis and the tricarboxylic acid cycle, and that these were more marked in glumes than leaves. Further, important differences are described on responsiveness of flag leaves and ears to e(CO2) on genes linked with carbon and nitrogen metabolism. These findings provide information about the impact of e(CO2) on ear development during the grain filling stage and are significant for understanding the effects of increasing (CO2) on crop yield.This research was supported by the Spanish Innovation and Universities Ministry (PCIN-2017-007) and the Ministry of Education and Sciences (grants PID2019-107154RB-100; PID2019-110445RB-100). RV was the recipient of an FPI fellowship from the Spanish Ministry of Economy and Competitiveness (BES-2010-031029). RS-B was supported by the Juan de la Cierva program JDC-Formación (FJCI-2016-28164)

New avenues for increasing yield and stability in C3 cereals: exploring ear photosynthesis

Small grain cereals such as wheat, rice and barley are among the most important crops worldwide. Any attempt to increase crop productivity and stability through breeding implies developing new strategies for plant phenotyping, including defining ideotype attributes for selection. Recently, the role of non-foliar photosynthetic organs, particularly the inflorescences, has received increasing attention. For example, ear photosynthesis has been reported to be a major contributor to grain filling in wheat and barley under stress and good agronomic conditions. This review provides an overview of the particular characteristics of the ear that makes this photosynthetic organ better adapted to grain filling than the flag leaf and revises potential metabolic and molecular traits that merit further research as targets for cereal improvement. Currently, the absence of high-throughput phenotyping methods limits the inclusion of ear photosynthesis in the breeding agenda. In this regard, a number of different approaches are presented

Carbon Isotope Composition and the NDVI as Phenotyping Approaches for Drought Adaptation in Durum Wheat: Beyond Trait Selection

High-throughput phenotyping platforms provide valuable opportunities to investigate biomass and drought-adaptive traits. We explored the capacity of traits associated with drought adaptation such as aerial measurements of the Normalized Difference Vegetation Index (NDVI) and carbon isotope composition (δ13C) determined at the leaf level to predict genetic variation in biomass. A panel of 248 elite durum wheat accessions was grown at the Maricopa Phenotyping platform (US) under well-watered conditions until anthesis, and then irrigation was stopped and plot biomass was harvested about three weeks later. Globally, the δ13C values increased from the first to the second sampling date, in keeping with the imposition of progressive water stress. Additionally, δ13C was negatively correlated with final biomass, and the correlation increased at the second sampling, suggesting that accessions with lower water-use efficiency maintained better water status and, thus, performed better. Flowering time affected NDVI predictions of biomass, revealing the importance of developmental stage when measuring the NDVI and the effect that phenology has on its accuracy when monitoring genotypic adaptation to specific environments. The results indicate that in addition to choosing the optimal phenotypic traits, the time at which they are assessed, and avoiding a wide genotypic range in phenology is crucial

The nitrogen contribution of different plant parts to wheat grains: exploring genotype, water, and nitrogen effects

The flag leaf has been traditionally considered as the main contributor to grain nitrogen. However, during the reproductive stage, other organs besides the flag leaf may supply nitrogen to developing grains. Therefore, the contribution of the ear and other organs to the nitrogen supplied to the growing grains remains unclear. It is important to develop phenotypic tools to assess the relative contribution of different plant parts to the N accumulated in the grains of wheat which may helps to develop genotypes that use N more efficiently. We studied the effect of growing conditions (different levels of water and nitrogen in the field) on the nitrogen contribution of the spike and different vegetative organs of the plant to the grains. The natural abundance of 15 δ N and total N content in the flag blade, peduncle, whole spike, glumes and awns were compared to the 15 δ N and total N in mature grains to trace the origin of nitrogen redistribution to the grains. The 15 δ N and total N content of the different plant parts correlated positively with the 15 δ N and total N content of mature grains suggesting that all organs may contribute a portion of their N content to the grains. The potential contribution of the flag blade to grain N increased (by 46%) as the growing conditions improved, whereas the potential contribution of the glumes plus awns and the peduncle increased (46 and 31%, respectively) as water and nitrogen stress increased. In general, potential contribution of the ear providing N to growing grains was similar (42%) than that of the vegetative parts of the plants (30-40%), regardless of the growing conditions. Thus, the potential ear N content could be a positive trait for plant phenotyping, especially under water and nitrogen limiting conditions. In that sense, genotypic variability existed at least between old (tall) and modern (semidwarf) cultivars, with the ear from modern genotypes exhibiting less relative contribution to the total grain N. The combined use of 15 δ N and N content may be used as an affordable tool to assess the relative contribution of different plant parts to the grain N in wheat

The Hydrogen isotope composition delta H-2 reflects plant performance

The stable carbon (δ13C) and oxygen (δ18O) isotope compositions in plant matter reflect photosynthetic and transpirative conditions in plants, respectively. However, the nature of hydrogen isotope composition (δ2H) and what it reflects of plant performance is poorly understood. Using durum wheat (Triticum turgidum var durum), this study evaluated the effect of different water and nitrogen growing field conditions on transpiration and how this effect influenced the performance of δ2H in autotrophic (flag leaf), mixotrophic (ears), and heterotrophic (grains and roots) organs. Moreover, δ2H was compared with the δ13C and δ18O in the same organs. Isotope compositions were analyzed in dry matter, the water-soluble fraction, and in water from different tissues of a set of genotypes. Similar to δ13C, the δ2H correlated negatively with stomatal conductance, whereas no correlation was observed for δ18O. Moreover, δ2H was not only affected by changes in transpiration but also by photosynthetic reactions, probably as a consequence of NADPH formation in autotrophic organs. Compared with the δ2H of stem water, plant δ2H was strongly diminished in photosynthetic organs such as the flag leaves, whereas it strongly increased in heterotrophic organs such as grains and roots. In heterotrophic organs, δ2H was associated with postphotosynthetic effects because there are several processes that lead to 2H-enrichment of carbohydrates. In summary, δ2H exhibited specific features that inform about the water conditions of the wheat crop, together with the photosynthetic characteristics of the plant part considered. Moreover, correlations of δ2H with grain yield illustrate that this isotope can be used to assess plant performance under different growing conditions