Álvaro Sanz-Sáez

Harvest index, a parameter conditioning responsiveness of wheat plants to elevated CO2

The expansion of the world’s population requires the development of high production agriculture. For this purpose, it is essential to identify target points conditioning crop responsiveness to predicted [CO2]. The aim of this study was to determine the relevance of ear sink strength in leaf protein and metabolomic profiles and its implications in photosynthetic activity and yield of durum wheat plants exposed to elevated [CO2]. For this purpose, a genotype with high harvest index (HI) (Triticum durum var. Sula) and another with low HI (Triticum durum var. Blanqueta) were exposed to elevated [CO2] (700 µmol mol–1 versus 400 µmol mol–1 CO2) in CO2 greenhouses. The obtained data highlighted that elevated [CO2] only increased plant growth in the genotype with the largest HI; Sula. Gas exchange analyses revealed that although exposure to 700 µmol mol–1 depleted Rubisco content, Sula was capable of increasing the light-saturated rate of CO2 assimilation (Asat) whereas, in Blanqueta, the carbohydrate imbalance induced the down-regulation of Asat. The specific depletion of Rubisco in both genotypes under elevated [CO2], together with the enhancement of other proteins in the Calvin cycle, revealed that there was a redistribution of N from Rubisco towards RuBP regeneration. Moreover, the down-regulation of N, NO3 –, amino acid, and organic acid content, together with the depletion of proteins involved in amino acid synthesis that was detected in Blanqueta grown at 700 µmol mol–1 CO2, revealed that inhibition of N assimilation was involved in the carbohydrate imbalance and consequently with the down-regulation of photosynthesis and growth in these plants.

Photosynthetic down-regulation under elevated CO2 exposure can be prevented by nitrogen supply in nodulated alfalfa.

Increasing atmospheric CO₂ concentrations are expected to enhance plant photosynthesis and yield. Nevertheless, after long-term exposure, plants acclimate and show a reduction in photosynthetic activity (called down-regulation), which may cause a reduction in potential yield. Some authors suggest that down-regulation is related to nutrient availability, and more specifically, to an insufficient plant C sink strength caused by limited N supply. In this paper, we tested whether or not N availability prevents down-regulation of photosynthesis in nodulated alfalfa plants (Medicago sativa L.). To do so, we examined the effect of the addition of different levels of NH₄NO₃ (0, 10, and 15 mM) to 30-day-old nodulated alfalfa plants exposed to ambient (approximately 400 μmol mol⁻¹) or elevated CO₂ (700 μmol mol⁻¹) during a period of 1 month in growth chambers. After 2 weeks of exposure to elevated CO₂, no significant differences were observed in plant growth or photosynthesis rates. After 4 weeks of treatment, exclusively N₂ fixing alfalfa plants (0 mM NH₄NO₃) showed significant decreases in photosynthesis and Vc(max). Photosynthetic down-regulation of these plants was caused by the C/N imbalance as reflected by the carbohydrate and N data. On the other hand, plants supplied with 15 mM NH₄NO₃ grown under elevated CO₂ maintained high photosynthetic rates owing to their superior C/N adjustment. The intermediate N treatment, 10 mM NH₄NO₃, also showed photosynthetic down-regulation, but to a lesser degree than with 0 mM treatment. The present study clearly shows that external N supply can reduce or even avoid acclimation of photosynthesis to elevated CO₂ as a consequence of the increase in C sink strength associated with N availability.

Carbon balance, partitioning and photosynthetic acclimation in fruit-bearing grapevine (Vitis vinifera L. cv. Tempranillo) grown under simulated climate change (elevated CO2, elevated temperature and moderate drought) scenarios in temperature gradient greenhouses

Although plant performance under elevated CO2 has been extensively studied in the past little is known about photosynthetic performance changing simultaneously CO2, water availability and temperature conditions. Moreover, despite of its relevancy in crop responsiveness to elevated CO2 conditions, plant level C balance is a topic that, comparatively, has received little attention. In order to test responsiveness of grapevine photosynthetic apparatus to predicted climate change conditions, grapevine (Vitis vinifera L. cv. Tempranillo) fruit-bearing cuttings were exposed to different CO2 (elevated, 700ppm vs. ambient, ca. 400ppm), temperature (ambient vs. elevated, ambient +4°C) and irrigation levels (partial vs. full irrigation). Carbon balance was followed monitoring net photosynthesis (AN, C gain), respiration (RD) and photorespiration (RL) (C losses). Modification of environment (13)C isotopic composition (δ(13)C) under elevated CO2 (from -10.30 to -24.93‰) enabled the further characterization of C partitioning into roots, cuttings, shoots, petioles, leaves, rachides and berries. Irrespective of irrigation level and temperature, exposure to elevated CO2 induced photosynthetic acclimation of plants. C/N imbalance reflected the inability of plants grown at 700ppm CO2 to develop strong C sinks. Partitioning of labeled C to storage organs (main stem and roots) did not avoid accumulation of labeled photoassimilates in leaves, affecting negatively Rubisco carboxylation activity. The study also revealed that, after 20 days of treatment, no oxidative damage to chlorophylls or carotenoids was observed, suggesting a protective role of CO2 either at current or elevated temperatures against the adverse effect of water stress.

Physiological traits contributed to the recent increase in yield potential of winter wheat from Henan Province, China

This experiment aims to test the traits responsible for the increase in yield potential of winter wheat released in Henan Province, China. Seven established cultivars released in the last 20 years and three advanced lines were assayed. The results showed that grain yield was positively correlated with harvest index (HI), kernel number per square meter, and aboveground biomass. In addition, the HI and aboveground biomass showed an increasing trend with the year of release. Therefore, we can conclude that bread wheat breeding advances during recent decades in Henan Province, China, have been achieved through an increase in HI, kernel number per square meter, and aboveground biomass. A higher δ(13)C seems also to be involved in these advances, which suggests a progressive improvement in constitutive water use efficiency not associated with a trend towards lower stomatal conductance in the most recent genotypes. However, genetic advance does not appear related to changes in photosynthesis rates on area basis when measured in the flag leaf or the spike, but only to a higher, whole-spike photosynthesis. Results also indirectly support the concept that under potential yield conditions, the spike contributed more than the flag leaf to kernel formation.

Alfalfa forage digestibility, quality and yield under future climate change scenarios vary with Sinorhizobium meliloti strain

Elevated CO(2) may decrease alfalfa forage quality and in vitro digestibility through a drop in crude protein and an enhancement of fibre content. The aim of the present study was to analyse the effect of elevated CO(2), elevated temperature and Sinorhizobium meliloti strains (102F78, 102F34 and 1032 GMI) on alfalfa yield, forage quality and in vitro dry matter digestibility. This objective is in line with the selection of S. meliloti strains in order to maintain high forage yield and quality under future climate conditions. Plants inoculated with the 102F34 strain showed more DM production than those inoculated with 1032GMI; however, these strains did not show significant differences with 102F78 plants. Neutral or acid detergent fibres were not enhanced in plants inoculated with the 102F34 strain under elevated CO(2) or temperature and hence, in vitro dry matter digestibility was unaffected. Crude protein content, an indicator of forage quality, was negatively related to shoot yield. Plants inoculated with 102F78 showed a similar shoot yield to those inoculated with 102F34, but had higher crude protein content at elevated CO(2) and temperature. Under these climate change conditions, 102F78 inoculated plants produced higher quality forage. However, the higher digestibility of plants inoculated with the 102F34 strain under any CO(2) or temperature conditions makes them more suitable for growing under climate change conditions. In general, elevated CO(2) in combination with high temperature (Climate Change scenario) reduced IVDMD and CP content and enhanced fibre content, which means that animal production will be negatively affected.

Photosynthesis, N2 fixation and taproot reserves during the cutting regrowth cycle of alfalfa under elevated CO2 and temperature

Future climatic conditions, including rising atmospheric CO(2) and temperature may increase photosynthesis and, consequently, plant production. A larger knowledge of legume performance under the predicted growth conditions will be crucial for safeguarding crop management and extending the area under cultivation with these plants in the near future. N(2) fixation is a key process conditioning plant responsiveness to varying growth conditions. Moreover, it is likely to increase under future environments, due to the higher photosynthate availability, as a consequence of the higher growth rate under elevated CO(2). However, as described in the literature, photosynthesis performance is frequently down-regulated (acclimated) under long-term exposure to CO(2), especially when affected by stressful temperature and water availability conditions. As growth responses to elevated CO(2) are dependent on sink-source status, it is generally accepted that down-regulation occurs in situations with insufficient plant C sink capacity. Alfalfa management involves the cutting of shoots, which alters the source-sink relationship and thus the photosynthetic behaviour. As the growth rate decreases at the end of the pre-cut vegetative growth period, nodulated alfalfa plants show photosynthetic down-regulation, but during regrowth following defoliation, acclimation to elevated CO(2) disappears. The shoot harvest also leads to a drop in mineral N uptake and C translocation to the roots, resulting in a reduction in N(2) fixation due to the dependence on photosynthate supply to support nodule function. Therefore, the production of new shoots during the first days following cutting requires the utilization of reduced C and N compounds that have been stored previously in reserve organs. The stored reserves are mediated by phytohormones such as methyl jasmonate and abscisic acid and in situations where water stress reduces shoot production this potentially enables the enhancement of taproot protein levels in nodulated alfalfa, which may lead to these plants being in better condition in the following cut/regrowth cycle. Furthering our knowledge of legume performance under predicted climate change conditions will be crucial for the development of varieties with better adaptation that will achieve greater and more efficient production values. Furthermore, for this purpose it will be necessary to improve existing methodologies and create new ones for phenotype characterization. Such knowledge will provide key information for future plant breeding programs.

Differential CO2 effect on primary carbon metabolism of flag leaves in durum wheat (Triticum durum Desf.).

C sink/source balance and N assimilation have been identified as target processes conditioning crop responsiveness to elevated CO2 . However, little is known about phenology-driven modifications of C and N primary metabolism at elevated CO2 in cereals such as wheat. Here, we examined the differential effect of elevated CO2 at two development stages (onset of flowering, onset of grain filling) in durum wheat (Triticum durum, var. Sula) using physiological measurements (photosynthesis, isotopes), metabolomics, proteomics and (15) N labelling. Our results show that growth at elevated CO2 was accompanied by photosynthetic acclimation through a lower internal (mesophyll) conductance but no significant effect on Rubisco content, maximal carboxylation or electron transfer. Growth at elevated CO2 altered photosynthate export and tended to accelerate leaf N remobilization, which was visible for several proteins and amino acids, as well as lysine degradation metabolism. However, grain biomass produced at elevated CO2 was larger and less N rich, suggesting that nitrogen use efficiency rather than photosynthesis is an important target for improvement, even in good CO2 -responsive cultivars.

Photosynthetic and Molecular Markers of CO2-mediated Photosynthetic Downregulation in Nodulated Alfalfa

Elevated CO₂ leads to a decrease in potential net photosynthesis in long-term experiments and thus to a reduction in potential growth. This process is known as photosynthetic downregulation. There is no agreement on the definition of which parameters are the most sensitive for detecting CO₂ acclimation. In order to investigate the most sensitive photosynthetic and molecular markers of CO₂ acclimation, the effects of elevated CO₂, and associated elevated temperature were analyzed in alfalfa plants inoculated with different Sinorhizobium meliloti strains. Plants (Medicago sativa L. cv. Aragón) were grown in summer or autumn in temperature gradient greenhouses (TGG). At the end of the experiment, all plants showed acclimation in both seasons, especially under elevated summer temperatures. This was probably due to the lower nitrogen (N) availability caused by decreased N₂-fixation under higher temperatures. Photosynthesis measured at growth CO₂ concentration, rubisco in vitro activity and maximum rate of carboxylation were the most sensitive parameters for detecting downregulation. Severe acclimation was also related with decreases in leaf nitrogen content associated with declines in rubisco content (large and small subunits) and activity that resulted in a drop in photosynthesis. Despite the sensitivity of rubisco content as a marker of acclimation, it was not coordinated with gene expression, possibly due to a lag between gene transcription and protein translation.

Harvest index combined with impaired N availability constrains the responsiveness of durum wheat to elevated CO2 concentration and terminal water stress

Despite its relevance, few studies to date have analysed the role of harvest index (HI) in the responsiveness of wheat (Triticum spp.) to elevated CO2 concentration ([CO2]) under limited water availability. The goal of the present work was to characterise the role of HI in the physiological responsiveness of durum wheat (Triticum durum Desf.) exposed to elevated [CO2] and terminal (i.e. during grain filling) water stress. For this purpose, the performance of wheat plants with high versus low HI (cvv. Sula and Blanqueta, respectively) was assessed under elevated [CO2] (700μmolmol-1 vs 400μmolmol-1 CO2) and terminal water stress (imposed after ear emergence) in CO2 greenhouses. Leaf carbohydrate build-up combined with limitations in CO2 diffusion (in droughted plants) limited the responsiveness to elevated [CO2] in both cultivars. Elevated [CO2] only increased wheat yield in fully watered Sula plants, where its larger HI prevented an elevated accumulation of total nonstructural carbohydrates. It is likely that the putative shortened grain filling period in plants exposed to water stress also limited the responsiveness of plants to elevated [CO2]. In summary, our study showed that even under optimal water availability conditions, only plants with a high HI responded to elevated [CO2] with increased plant growth, and that terminal drought constrained the responsiveness of wheat plants to elevated [CO2].

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.