M.C. Antolín

Effect of water stress on photosynthetic activity in the Medicago-Rhizobium-Glomus symbiosis

Abstract CO 2 exchange rate (CER), internal CO 2 concentration (Ci), photosynthetical phosphorus use efficiency (PPUE), nodule activity (ARA) and growth parameters were compared in mycorrhizal nodulated (VAM) and P-compensated nodulated alfalfa plants, under both well watered and drought acclimated conditions. Leaf area ratio significantly decreased in P-fertilized plants in response to drought treatment whereas mycorrhizal plants maintained rather constant values. As stress progressed, VAM plants developed lower root-shoot and higher leaf area ratios than non-VAM. Total dry weight decreased for both treatments in response to drought, but reduction was more pronounced in P-fertilized plants. Under watered conditions, CER and PPUE were approximately 30% and 55% higher respectively in VAM than in non-VAM plants while specific nodule activity was similar in both groups of plants. Throughout drought, the three parameters (CER, PPUE and ARA) maintained significantly higher values in myocorrhizal plants. Internal CO 2 concentration was always higher in P-compensated than in VAM plants. Results show that under water stress the tripartite symbiosis maintains higher values of photosynthetic and nodule activities than non-myocorrhizal plants suggesting enhanced drought tolerance by mycorrhizal condition.

Influence of arbuscular mycorrhizae and Rhizobium on nutrient content and water relations in drought stressed alfalfa

The objective of this research was to study the effect of drought on nutrient content and leaf water status in alfalfa (Medicago sativa L. cv Aragón) plants inoculated with a mycorrhizal fungus and/or Rhizobium compared with noninoculated ones. The four treatments were: a) plants inoculated with Glomus fasciculatum and Rhizobium meliloti 102 F51 strain, (MR); b) plants inoculated with R. meliloti only (R); c) plants with G. fasciculatum only (M); and d) noninoculated plants (N). Nonmycorrhizal plants were supplemented with phosphorus and nonnodulated ones with nitrogen to achieve similar size and nutrient content in all treatments. Plants were drought stressed using two cycles of moisture stress and recovery. The components of total leaf water potential (osmotic and pressure potentials at full turgor), percentage of apoplastic water volume and the bulk modulus of elasticity of leaf tissue were determined. Macronutrient (N, P, K, Ca, S and Mg) and micronutrient (Co, Mo, Zn, Mn, Cu, Na, Fe and B) content per plant were also measured. Leaves of N and R plants had decreased osmotic potentials and increased pressure potentials at full turgor, with no changes either in the bulk modulus of elasticity or the percentage of apoplastic water upon drought conditions. By contrast, M and MR leaves did not vary in osmotic and turgor potentials under drought stress but had increased apoplastic water volume and cell elasticity (lowering bulk modulus). Drought stress decreased nutrient content of leaves and roots of noninoculated plants. R plants showed a decrease in nutrient content of leaves but maintained some micronutrients in roots. Leaves of M plants were similar in content of nutrients to N plants. However, roots of M and MR plants had significantly lower nutrient content. Results indicate an enhancement of nutrient content in mycorrhizal alfalfa plants during drought that affected leaf water relations during drought stress.

Influence of arbuscular mycorrhizae and Rhizobium on free polyamines and proline levels in water-stressed alfalfa

Summary The objective of this research was to study the effect of drought on polyamine and proline levels in alfalfa ( Medicago sativa L. cv. Aragon) plants inoculated with a mycorrhizal fungus and/or Rhizobium compared with non-inoculated ones. The four treatments were: a) plants inoculated with Glomus fasciculatum (Taxter sensu Gerd.) Gerdemann and Trappe and Rhizobium meliloti 102 F51 strain (MR), b) plants inoculated with Rhizobium only (R), c) plants inoculated with Glomus only (M), and d) non-inoculated plants (N). Plants were drought stressed during two cycles of moisture stress and recovery. Although proline concentrations increased and free polyamine (spermidine and spermine) contents decreased in leaves and roots of alfalfa under water stress, symbiotic R, M and MR plants maintained higher free polyamine concentrations than non-symbiotic N ones. Results suggest that symbiotic alfalfa plants are better adapted than non-symbiotic ones to cope with water deficit.

Effects of temporary drought on nitrate-fed and nitrogen-fixing alfalfa plants

Abstract The effect of temporary drought on growth, carbon exchange and solute accumulation has been examined in alfalfa plants dependent on either N2 or nitrate. Plants were subjected to cyclical moderate or severe drought (drought/recovery). Growth parameters, photosynthetic rate (Pn), leaf conductance to water vapour (gw), chlorophyll content, and solute accumulation were determined. Growth decreased markedly under water deficit, but no significant differences between either groups of plants were found. Nitrogen-fixing plants developed higher root/shoot ratios maintaining larger leaves with increased specific leaf area and greater chlorophyll content than nitrate-fed ones. Leaf conductance and net photosynthetic rate declined simultaneously with the drought treatments in both groups of plants; however, N2 fixing plants retained higher Pn and gw values than nitrate-fed plants at lower RWC. Upon rewatering, a considerable stomatal closure remained in nitrate-fed plants. Drought treatment induced an increase in solute concentrations, mainly potassium, especially important in nitrate-fed plants. The interactions between the type of N nutrition and drought tolerance in alfalfa plants during temporary drought are discussed.

Reproductive response of two morphologically different pea cultivars to drought

Abstract Water use by semi-leafless peas ( Pisum sativum L.) is usually less than that of conventional peas because of their reduced surface leaf area, suggesting that semi-leafless peas would be less sensitive to drought because drought develops later. This work aimed to study the reproductive response of peas cv. Solara (semi-leafless) and cv. Frilene (conventional) subjected to similar controlled soil drought during the critical period occurring between flowering and initial seed filling. Plants were subjected to drought by watering with a fraction of water used in the evapotranspiration of control plants. Soil, pod and seed water contents, leaf water status parameters, dry matter (DM) partitioning, seed yield, yield components and water use efficiency (WUE) were measured. Although soil water content decreased in a similar way in both cultivars, leaf Ψ w and RWC only decreased significantly in Solara. Well-watered Frilene plants produced higher shoot and pod DM, but lower seed DM. Well-watered Solara plants produced lower pod DM and higher seed DM than Frilene. Under drought, Frilene increased partitioning of total plant DM to vegetative organs, particularly roots, and decreased DM allocation to pods and seeds increasing flower abortion. By contrast, droughted Solara interrupted vegetative growth and increased leaf senescence but maintained similar partitioning of total plant DM to pods and seeds as in well-watered conditions. For both cultivars there was a close relationship between the percentage of total DM partitioned into seeds and WUE y (water use efficiency on seed yield basis). Results demonstrate that when plants suffered the same level of drought in the soil, the reproductive response of the two cultivars was linked to differences in their WUE.

Methodological advances: using greenhouses to simulate climate change scenarios.

Human activities are increasing atmospheric CO2 concentration and temperature. Related to this global warming, periods of low water availability are also expected to increase. Thus, CO2 concentration, temperature and water availability are three of the main factors related to climate change that potentially may influence crops and ecosystems. In this report, we describe the use of growth chamber - greenhouses (GCG) and temperature gradient greenhouses (TGG) to simulate climate change scenarios and to investigate possible plant responses. In the GCG, CO2 concentration, temperature and water availability are set to act simultaneously, enabling comparison of a current situation with a future one. Other characteristics of the GCG are a relative large space of work, fine control of the relative humidity, plant fertirrigation and the possibility of light supplementation, within the photosynthetic active radiation (PAR) region and/or with ultraviolet-B (UV-B) light. In the TGG, the three above-mentioned factors can act independently or in interaction, enabling more mechanistic studies aimed to elucidate the limiting factor(s) responsible for a given plant response. Examples of experiments, including some aimed to study photosynthetic acclimation, a phenomenon that leads to decreased photosynthetic capacity under long-term exposures to elevated CO2, using GCG and TGG are reported.

Growth, photosynthetic acclimation and yield quality in legumes under climate change simulations: An updated survey

Continued emissions of CO2, derived from human activities, increase atmospheric CO2 concentration. The CO2 rise stimulates plant growth and affects yield quality. Effects of elevated CO2 on legume quality depend on interactions with N2-fixing bacteria and mycorrhizal fungi. Growth at elevated CO2 increases photosynthesis under short-term exposures in C3 species. Under long-term exposures, however, plants generally acclimate to elevated CO2 decreasing their photosynthetic capacity. An updated survey of the literature indicates that a key factor, perhaps the most important, that characteristically influences this phenomenon, its occurrence and extent, is the plant source-sink balance. In legumes, the ability of exchanging C for N at nodule level with the N2-fixing symbionts creates an extra C sink that avoids the occurrence of photosynthetic acclimation. Arbuscular mycorrhizal fungi colonizing roots may also result in increased C sink, preventing photosynthetic acclimation. Defoliation (Anthyllis vulneraria, simulated grazing) or shoot cutting (alfalfa, usual management as forage) largely increases root/shoot ratio. During re-growth at elevated CO2, new shoots growth and nodule respiration function as strong C sinks that counteracts photosynthetic acclimation. In the presence of some limiting factor, the legumes response to elevated CO2 is weakened showing photosynthetic acclimation. This survey has identified limiting factors that include an insufficient N supply from bacterial strains, nutrient-poor soils, low P supply, excess temperature affecting photosynthesis and/or nodule activity, a genetically determined low nodulation capacity, an inability of species or varieties to increase growth (and therefore C sink) at elevated CO2 and a plant phenological state or season when plant growth is stopped.

The Role of Plant Size and Nutrient Concentrations in Associations between Medicago, and Rhizobium and/or Glomus

The aim of this research was to carry out a critical study of the method of obtaining size equivalence between non-symbiotic alfalfa and alfalfa associated with Glomus and/or Rhizobium by applying fixed addition rates of nutrients to the non-symbiotic controls. The experimental design included three nutrient response curves in which the levels of added phosphorus and/or nitrogen were constant during the whole plant growth process: 1) a phosphorus response curve, in order to compare the growth of double symbiotic plants with that of only-Rhizobium inoculated ones; 2) a nitrogen response curve, that consisted of a comparison between the growth of double symbiotic alfalfa and four treatments associated only with Glomus; 3) a phosphorus and nitrogen response curve, to compare the growth of non-inoculated alfalfa with that of double symbiotic plants. Although similar size was achieved among some treatments at harvest, shoot growth over time and nutrient concentrations in tissues differed, indicating that growth equivalence did not mean functional equivalence. A second experimental design was performed taking into account the establishment of microsymbionts for determining the adequate moment to add supplemental phosphorus and/or nitrogen. It included four treatments: a) double symbiotic plants (MR); b) plants inoculated with Rhizobium only (R); c) plants inoculated with Glomus only (M), and d) non-inoculated plants (N). Great similarity in terms of plant growth and nutrient contents in tissues were obtained. Moreover, symbiotic plants were able to produce similar dry matter than non-symbiotic ones under P and N limitations.

Effects of nitrogen source and water availability on stem carbohydrates and cellulosic bioethanol traits of alfalfa plants

Symbiotic association of legumes with rhizobia frequently results in higher photosynthesis and soluble carbohydrates in comparison with nitrate-fed plants, which might improve its potential for biomass conversion into bioethanol. A greenhouse experiment was conducted to examine the effects of nitrogen source and water availability on stem characteristics and on relationships between carbohydrates, phenolic metabolism activity and cell wall composition in alfalfa (Medicago sativa L. cv. Aragón). The experiment included three treatments: (1) plants fed with ammonium nitrate (AN); (2) plants inoculated with rhizobia (R); and (3) plants inoculated with rhizobia and amended with sewage sludge (RS). Two levels of irrigation were imposed: (1) well-watered and (2) drought stress. Under well-watered conditions, nitrogen-fixing plants have increased photosynthesis and stem fermentable carbohydrate concentrations, which result in higher potential for biomass conversion to bioethanol than in AN plants. The latter had higher lignin due to enhanced activities of phenolic metabolism-related enzymes. Under drought conditions, the potential for bioethanol conversion decreased to a similar level in all treatments. Drought-stressed nitrogen-fixing plants have high concentrations of fermentable carbohydrates and cell wall cellulose, but ammonium nitrate-fed plants produced higher plant and stem biomass, which might compensate the decreasing stem carbohydrates and cellulose concentrations.