Response of nutrient cycles of an old-growth montane forest in Ecuador to experimental low-level nutrient amendments

Atmospheric nitrogen (N) and phosphorus (P) depositions are expected to increase in the tropicsrnas a consequence of increasing human activities in the next decades. Furthermore, a possiblernshortened El Niño Southern Oscillation cycle might come along with more frequent calcium (Ca)rndepositions on the eastern slope of the Ecuadorian Andes originating from Saharan dust. It isrncrucial to understand the response of the old-growth montane forest in Ecuador to increasedrnnutrient deposition to predict the further development of this megadiverse ecosystem.rnI studied experimental additions of N, P, N+P and Ca to the forest and an untreatedrncontrol, all in a fourfold replicated randomized block design. These experiments were conductedrnin the framework of a collaborative research effort, the NUtrient Manipulation EXperimentrn(NUMEX). I collected litter leachate, mineral soil solution (0.15 and 0.30 m depths), throughfallrnand fine litterfall samples and determined N, P and Ca concentrations and fluxes. This approachrnalso allowed me to assess whether N, P and/or Ca are limiting nutrients for forest growth.rnFurthermore, I evaluated the response of fine root biomass, leaf area index, leaf area and specificrnleaf area, tree diameter growth and basal area increment contributed from a cooperating group inrnthe Ca applied and control treatments.rnDuring the observation period of 16 months after the first fertilizer application, less thanrn10, 1 and 5% of the applied N, P and Ca, respectively, leached below the organic layer whichrncontained almost all roots but no significant leaching losses occurred to the deeper mineral soil.rnDeposited N, P and Ca from the atmosphere in dry and wet form were, on balance, retained in therncanopy in the control treatment. Retention of N, P and Ca in the canopy in their respectiverntreatments was reduced resulting in higher concentrations and fluxes of N, P and Ca inrnthroughfall and litterfall. Up to 2.5% of the applied N and 2% of the applied P and Ca werernrecycled to the soil with throughfall. Fluxes of N, P and Ca in throughfall+litterfall were higher inrnthe fertilized treatments than in the control; up to 20, 5 and 25% of the applied N, P and Ca,rnrespectively, were recycled to the soil with throughfall+litterfall.rnIn the Ca-applied plots, fine root biomass decreased significantly. Also the leaf area of thernfour most common tree species tended to decrease and the specific leaf area increasedrnsignificantly in Graffenrieda emarginata Triana, the most common tree species in the study area.rnThese changes are known plant responses to reduced nutrient stress. Reduced aluminium (Al)rntoxicity as an explanation of the Ca effect was unlikely, because of almost complete organocomplexationrnof Al and molar Ca:Al concentration ratios in solution above the toxicity threshold.rnThe results suggest that N, P and Ca co-limit the forest ecosystem functioning in thernnorthern Andean montane forests in line with recent assumptions in which different ecosystemrncompartments and even different phenological stages may show different nutrient limitationsrn(Kaspari et al. 2008). I conclude that (1) the expected elevated N and P deposition will bernretained in the ecosystem, at least in the short term and hence, quality of river water will not bernendangered and (2) increased Ca input will reduce nutrient stress of the forest.

Characterization of the rice PHO1 gene family reveals a key role for OsPHO1;2 in phosphate homeostasis and the evolution of a distinct clade in dicotyledons.

Phosphate homeostasis was studied in a monocotyledonous model plant through the characterization of the PHO1 gene family in rice (Oryza sativa). Bioinformatics and phylogenetic analysis showed that the rice genome has three PHO1 homologs, which cluster with the Arabidopsis (Arabidopsis thaliana) AtPHO1 and AtPHO1;H1, the only two genes known to be involved in root-to-shoot transfer of phosphate. In contrast to the Arabidopsis PHO1 gene family, all three rice PHO1 genes have a cis-natural antisense transcript located at the 5 ' end of the genes. Strand-specific quantitative reverse transcription-PCR analyses revealed distinct patterns of expression for sense and antisense transcripts for all three genes, both at the level of tissue expression and in response to nutrient stress. The most abundantly expressed gene was OsPHO1;2 in the roots, for both sense and antisense transcripts. However, while the OsPHO1;2 sense transcript was relatively stable under various nutrient deficiencies, the antisense transcript was highly induced by inorganic phosphate (Pi) deficiency. Characterization of Ospho1;1 and Ospho1;2 insertion mutants revealed that only Ospho1;2 mutants had defects in Pi homeostasis, namely strong reduction in Pi transfer from root to shoot, which was accompanied by low-shoot and high-root Pi. Our data identify OsPHO1;2 as playing a key role in the transfer of Pi from roots to shoots in rice, and indicate that this gene could be regulated by its cis-natural antisense transcripts. Furthermore, phylogenetic analysis of PHO1 homologs in monocotyledons and dicotyledons revealed the emergence of a distinct clade of PHO1 genes in dicotyledons, which include members having roles other than long-distance Pi transport.

Serological diagnosis of Chagas disease: evaluation and characterisation of a low cost antigen with high sensitivity and specificity

In spite of evident progress in the serology of Chagas disease, the requirement for new diagnostic antigens persists. We have evaluated different antigens obtained from Trypanosoma cruzi grown in medium rich in nutrients or under nutrient stress, autoclaved or sonicated and fractionated by differential centrifugation. The resulting antigens were evaluated for diagnosis of Chagas disease using ELISA. Immunofluorescence of the parasites demonstrated that nutrient stress induced changes in the distribution and density of antigens recognised by a pool of sera from experimentally infected mice. When evaluated using ELISA, it was evident that most fractions had good sensitivity but poor specificity. Surprisingly, the best specificity and sensitivity was observed with parasites cultured under nutrient stress and autoclaved. Furthermore this antigen had low cross reactivity with sera from other parasitic diseases, Leishmaniasis in particular. Western blot analysis demonstrated that autoclaving seems to non-specifically eliminate cross-reactive antigens. In conclusion, autoclaving epimastigotes of T. cruzi, after nutrient stress, allowed us to obtain an antigen that could be used in the serological diagnosis of Chagas disease.

Ecological distribution of Cyanophyceae in lotic ecosystems of São Paulo State

A hundred seventy two stream reaches from six distinct natural regions (parts of biomes or geological areas) in São Paulo State (Atlantic Rainforest, Cerrado, Hard Water regions, Northwest region, Subtropical Rainforest, and Tropical Rainforest) were searched for the most representative macroscopic Cyanophyceae of phytobenthic communities. Selected ecological parameters were analyzed in each stream segment: conductance, current velocity, oxygen saturation, pH, turbidity, and water temperature. Algal abundance was evaluated as percentage cover. Thirthy four cyanophyte taxa were identified and Phormidium retzii was the most widespread species throughout the State, occurring in all studied regions, except in hard water areas, and was negatively correlated to conductance. The subtropical Rainforest region presented the highest mean species richness per site, whereas the tropical Rainforest region had the highest abundance (percentage cover) of blue-green algae. Correlation tests revealed that conductance was significantly and negatively related to variations in abundance and richness of cyanophyte in streams of São Paulo State. This relationship probably reflects the ability of blue-green algae to grow at medium to low ion content and to take advantage under nutrient stress conditions.

Cultivo de Haematococcus pluvialis sob condições de estresse e em consórcio microbiano visando obter maior rendimento de astaxantina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Parâmetros hemetológicos da tilápia-do-Nilo: efeito da dieta suplementada com levedura e zinco e do estímulo pelo frio

Fundação de Apoio à Pesquisa do Estado de São Paulo (FAPESP)

Modelling growth dynamics and toxin production in Ostreopsis cf. ovata

The benthic dinoflagellate O. ovata represents a serious threat for human health and for the ecology of its blooming areas: thanks to its toxicity this microalga has been responsible for several cases of human intoxication and mass mortalities of benthic invertebrates. Although the large number of studies on this dinoflagellate, the mechanisms underpinning O. ovata growth and toxin production are still far to be fully understood. In this work we have enriched the dataset on this species by carrying out a new experiment on an Adriatic O. cf. ovata strain. Data from this experiment (named Beta) and from another comparable experiment previously conducted on the same strain (named Alpha), revealed some interesting aspects of this dinoflagellate: it is able to grow also in a condition of strong intracellular nutrient deficiency (C:P molar ratio > 400; C:N > 25), reaching extremely low values of chlorophyll-a to carbon ratio (0.0004). Was also found a significant inverse relationships (r > -0.7) between cellular toxin to carbon and cellular nutrient to carbon ratios of experiment Alpha. In the light of these result, we hypothesized that in O. cf. ovata nutrient-stress conditions (intended as intracellular nutrient deficiency) can cause: i) an increase in toxin production; ii) a strong decrease in chlorophyll-a synthesis; iii) a lowering of metabolism associated with the formation of a sort of resting stage. We then used a modelling approach to test and critically evaluate these hypotheses in a mechanistic way: newly developed formulation describing toxin production and fate, and ad hoc changes in the already existent formulations describing chlorophyll synthesis, rest respiration, and mortality, have been incorporated in a simplified version of the European Regional Seas Ecosystem Model (ERSEM), together with a new ad hoc parameterization. The adapted model was able to accurately reproduce many of the trends observed in the Alpha experiment, allowing us to support our hypotheses. Instead the simulations of the experiment Beta were not fully satisfying in quantitative terms. We explained this gap with the presumed different physiological behaviors between the algae of the two experiments, due to the different pre-experimental periods of acclimation: the model was not able to reproduce acclimation processes in its simulations of the experiment Beta. Thus we attempt to simulate the acclimation of the algae to nutrient-stress conditions by manual intervention on some parameters of nutrient-stress thresholds, but we received conflicting results. Further studies are required to shed light on this interesting aspect. In this work we also improve the range of applicability of a state of the art marine biogeochemical model (ERSEM) by implementing in it an ecological relevant process such as the production of toxic compounds.

Satellite-Detected Fluorescence Reveals Global Physiology of Ocean Phytoplankton

Phytoplankton photosynthesis links global ocean biology and climate-driven fluctuations in the physical environment. These interactions are largely expressed through changes in phytoplankton physiology, but physiological status has proven extremely challenging to characterize globally. Phytoplankton fluorescence does provide a rich source of physiological information long exploited in laboratory and field studies, and is now observed from space. Here we evaluate the physiological underpinnings of global variations in satellite-based phytoplankton chlorophyll fluorescence. The three dominant factors influencing fluorescence distributions are chlorophyll concentration, pigment packaging effects on light absorption, and light-dependent energy-quenching processes. After accounting for these three factors, resultant global distributions of quenching-corrected fluorescence quantum yields reveal a striking consistency with anticipated patterns of iron availability. High fluorescence quantum yields are typically found in low iron waters, while low quantum yields dominate regions where other environmental factors are most limiting to phytoplankton growth. Specific properties of photosynthetic membranes are discussed that provide a mechanistic view linking iron stress to satellite-detected fluorescence. Our results present satellite-based fluorescence as a valuable tool for evaluating nutrient stress predictions in ocean ecosystem models and give the first synoptic observational evidence that iron plays an important role in seasonal phytoplankton dynamics of the Indian Ocean. Satellite fluorescence may also provide a path for monitoring climate-phytoplankton physiology interactions and improving descriptions of phytoplankton light use efficiencies in ocean productivity models.

Carbon-Based Primary Productivity Modeling With Vertically Resolved Photoacclimation

Net primary production (NPP) is commonly modeled as a function of chlorophyll concentration (Chl), even though it has been long recognized that variability in intracellular chlorophyll content from light acclimation and nutrient stress confounds the relationship between Chl and phytoplankton biomass. It was suggested previously that satellite estimates of backscattering can be related to phytoplankton carbon biomass (C) under conditions of a conserved particle size distribution or a relatively stable relationship between C and total particulate organic carbon. Together, C and Chl can be used to describe physiological state (through variations in Chl:C ratios) and NPP. Here, we fully develop the carbon-based productivity model (CbPM) to include information on the subsurface light field and nitracline depths to parameterize photoacclimation and nutrient stress throughout the water column. This depth-resolved approach produces profiles of biological properties (Chl, C, NPP) that are broadly consistent with observations. The CbPM is validated using regional in situ data sets of irradiance-derived products, phytoplankton chlorophyll: carbon ratios, and measured NPP rates. CbPM-based distributions of global NPP are significantly different in both space and time from previous Chl-based estimates because of the distinction between biomass and physiological influences on global Chl fields. The new model yields annual, areally integrated water column production of similar to 52 Pg C a(-1) for the global oceans.

Regulation of Protein Degradation in the Heart by AMP-Activated Protein Kinase

The degradation of proteins by the ubiquitin proteasome system is essential for cellular homeostasis in the heart. An important regulator of metabolic homeostasis is AMP-activated protein kinase (AMPK). During nutrient deprivation, AMPK is activated and intracellular proteolysis is enhanced through the ubiquitin proteasome system (UPS). Whether AMPK plays a role in protein degradation through the UPS in the heart is not known. Here I present data in support of the hypothesis that AMPK transcriptionally regulates key players in the UPS, which, under extreme conditions can be detrimental to the heart. The ubiquitin ligases MAFbx /Atrogin-1 and MuRF1, key regulators of protein degradation, and AMPK activity are increased during nutrient deprivation. Pharmacologic and genetic activation of AMPK is sufficient for the induction of MAFbx/Atrogin-1 and MuRF1 in cardiomyocytes and in the heart in vivo. Comprehensive experiments demonstrate that the molecular mechanism by which AMPK regulates MuRF1 expression is through the transcription factor myocyte enhancer factor 2 (MEF2), which is involved in stress response and cardiomyocyte remodeling. MuRF1 is required for AMPK-mediated protein degradation through the UPS in cardiomyocytes. Consequently, the absence of MuRF1 during chronic fasting preserves cardiac function, possibly by limiting degradation of critical metabolic enzymes. Furthermore, during cardiac hypertrophy, chronic activation of AMPK also leads to cardiac dysfunction, possibly through enhanced protein degradation and metabolic dysregulation. Collectively, my findings demonstrate that AMPK regulates expression of ubiquitin ligases which are required for UPS-mediated protein degradation in the heart. Based on these results, I propose that specific metabolic signals may serve as modulators of intracellular protein degradation in the heart.

Experiment: Organic matter exudation by Emiliania huxleyi under simulated future ocean conditions

Emiliania huxleyi (strain B 92/11) was exposed to different nutrient supply, CO2 and temperature conditions in phosphorus controlled chemostats to investigate effects on organic carbon exudation and partitioning between the pools of particulate organic carbon (POC) and dissolved organic carbon (DOC). 14C incubation measurements for primary production (PP) and extracellular release (ER) were performed. Chemical analysis included the amount and composition of high molecular weight (>1 kDa) dissolved combined carbohydrates (HMW-dCCHO), particulate combined carbohydrates (pCCHO) and the carbon content of transparent exopolymer particles (TEP-C). Applied CO2 and temperature conditions were 300, 550 and 900 µatm pCO2 at 14 °C, and additionally 900 µatm pCO2 at 18 °C simulating a greenhouse ocean scenario. Enhanced nutrient stress by reducing the dilution rate (D) from D = 0.3 /d to D = 0.1 /d (D = µ) induced the strongest response in E. huxleyi. At µ = 0.3 /d, PP was significantly higher at elevated CO2 and temperature and DO14C production correlated to PO14C production in all treatments, resulting in similar percentages of extracellular release (PER; (DO14C production/PP) × 100) averaging 3.74 ± 0.94%. At µ = 0.1 /d, PO14C production decreased significantly, while exudation of DO14C increased. Thus, indicating a stronger partitioning from the particulate to the dissolved pool. Maximum PER of 16.3 ± 2.3% were observed at µ = 0.1 /d at elevated CO2 and temperature. While cell densities remained constant within each treatment and throughout the experiment, concentrations of HMW-dCCHO, pCCHO and TEP were generally higher under enhanced nutrient stress. At µ= 0.3 /d, pCCHO concentration increased significantly with elevated CO2 and temperature. At µ = 0.1 /d, the contribution (mol % C) of HMW-dCCHO to DOC was lower at elevated CO2 and temperature while pCCHO and TEP concentrations were higher. This was most pronounced under greenhouse conditions. Our findings suggest a stronger transformation of primary produced DOC into POC by coagulation of exudates under nutrient limitation. Our results further imply that elevated CO2 and temperature will increase exudation by E. huxleyi and may affect organic carbon partitioning in the ocean due to an enhanced transfer of HMW-dCCHO to TEP by aggregation processes.

Transgenically Enhanced Sorbitol Synthesis Facilitates Phloem Boron Transport and Increases Tolerance of Tobacco to Boron Deficiency1

The mobility of elements within plants contributes to a plant species' tolerance of nutrient deficiencies in the soil. The genetic manipulation of within-plant nutrient movement may therefore provide a means to enhance plant growth under conditions of variable soil nutrient availability. In these experiments tobacco (Nicotiana tabacum) was engineered to synthesize sorbitol, and the resultant effect on phloem mobility of boron (B) was determined. In contrast to wild-type tobacco, transgenic tobacco plants containing sorbitol exhibit a marked increase in within-plant B mobility and a resultant increase in plant growth and yield when grown with limited or interrupted soil B supply. Growth of transgenic tobacco could be maintained by reutilization of B present in mature tissues or from B supplied as a foliar application to mature leaves. In contrast, B present in mature leaves of control tobacco lines could not be used to provide the B requirements for new plant growth. 10B-labeling experiments verified that B is phloem mobile in transgenic tobacco but is immobile in control lines. These results demonstrate that the transgenic enhancement of within-plant nutrient mobility is a viable approach to improve plant tolerance of nutrient stress.

Transcriptome analysis of intraspecific competition in Arabidopsis thaliana reveals organ-specific signatures related to nutrient acquisition and general stress response pathways.

ABSTRACT: BACKGROUND: Plants are sessile and therefore have to perceive and adjust to changes in their environment. The presence of neighbours leads to a competitive situation where resources and space will be limited. Complex adaptive responses to such situation are poorly understood at the molecular level. RESULTS: Using microarrays, we analysed whole-genome expression changes in Arabidopsis thaliana plants subjected to intraspecific competition. The leaf and root transcriptome was strongly altered by competition. Differentially expressed genes were enriched in genes involved in nutrient deficiency (mainly N, P, K), perception of light quality, and responses to abiotic and biotic stresses. Interestingly, performance of the generalist insect Spodoptera littoralis on densely grown plants was significantly reduced, suggesting that plants under competition display enhanced resistance to herbivory. CONCLUSIONS: This study provides a comprehensive list of genes whose expression is affected by intraspecific competition in Arabidopsis. The outcome is a unique response that involves genes related to light, nutrient deficiency, abiotic stress, and defence responses.

Root Water Extraction under Combined Water and Osmotic Stress

Using a numerical implicit model for root water extraction by a single root in a symmetric radial flow problem, based on the Richards equation and the combined convection-dispersion equation, we investigated some aspects of the response of root water uptake to combined water and osmotic stress. The model implicitly incorporates the effect of simultaneous pressure head and osmotic head on root water uptake, and does not require additional assumptions (additive or multiplicative) to derive the combined effect of water and salt stress. Simulation results showed that relative transpiration equals relative matric flux potential, which is defined as the matric flux potential calculated with an osmotic pressure head-dependent lower bound of integration, divided by the matric flux potential at the onset of limiting hydraulic conditions. In the falling rate phase, the osmotic head near the root surface was shown to increase in time due to decreasing root water extraction rates, causing a more gradual decline of relative transpiration than with water stress alone. Results furthermore show that osmotic stress effects on uptake depend on pressure head or water content, allowing a refinement of the approach in which fixed reduction factors based on the electrical conductivity of the saturated soil solution extract are used. One of the consequences is that osmotic stress is predicted to occur in situations not predicted by the saturation extract analysis approach. It is also shown that this way of combining salinity and water as stressors yields results that are different from a purely multiplicative approach. An analytical steady state solution is presented to calculate the solute content at the root surface, and compared with the outputs of the numerical model. Using the analytical solution, a method has been developed to estimate relative transpiration as a function of system parameters, which are often already used in vadose zone models: potential transpiration rate, root length density, minimum root surface pressure head, and soil theta-h and K-h functions.

Zn uptake, physiological response and stress attenuation in mycorrhizal jack bean growing in soil with increasing Zn concentrations

The influence of arbuscular mycorrhizal fungi (AMF) inoculation on Canavalia ensiformis growth. nutrient and Zn uptake, and on some physiological parameters in response to increasing soil Zn concentrations was studied. Treatments were applied in seven replicates in a 2 x 4 factorial design, consisting of the inoculation or not with the AMF Glomus etunicatum, and the addition of Zn to soil at the concentrations of 0, 100, 300 and 900 mg kg(-1). AMF inoculation enhanced the accumulation of Zn in tissues and promoted biomass yields and root nodulation. Mycorrhizal plants exhibited relative tolerance to Zn up to 300 mg kg(-1) without exhibiting visual symptoms of toxicity, in contrast to non-mycorrhizal plants which exhibited a significant growth reduction at the same soil Zn concentration. The highest concentration of Zn added to soil was highly toxic to the plants. Leaves of plants grown in high Zn concentration exhibited a Zn-induced proline accumulation and also an increase in soluble amino acid contents; however proline contents were lower in mycorrhizal jack beans. Plants in association or not with the AMF exhibited marked differences in the foliar soluble amino acid profile and composition in response to Zn addition to soil. In general, Zn induced oxidative stress which could be verified by increased lipid peroxidation rates and changes in catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase activities. In summary, G. etunicatum was able to maintain an efficient symbiosis with jack bean plants in moderately contaminated Zn-soils, improving plant performance under those conditions, which is likely to be due to a combination of physiological and nutritional changes caused by the intimate relation between fungus and plant. The enhanced Zn uptake by AMF inoculated jack bean plants might be of interest for phytoremediation purposes. (C) 2009 Elsevier Ltd. All rights reserved.