Effect of mannitol and cold treatments on phosphate uptake and protein phosphorylation in Lemna minor (L.) plants

This report is aimed at elucidating the effect of mannitol and cold treatments on P uptake and protein phosphorylation in Lemna minor plants. Duckweed p lants were incu bated in the presence of [32P]or [33P]Pi in half-strength phosphate deprived E-medium under constant light regime for 1.5 h. Total plant protein extracts (pellet and supernatant) were then prepared and subjected to IEF x SDS-PAGE. To analyse the effect of the stresses on P uptake and protein labelling, Lemna minor plants were preincubated with 0.1, 0.5 mol · L-1 mannitol and at 4°C respectively, for 4 hours, before adding labelled orthophosphate. The results show that the general protein phosphorylation (including LHCII) is related to the level of P uptake. Radioactive phosphate incorporation is stimulated by a low concentration of mannitol (0.1 mol · L-1) but reduced by 0.5 mol · L-1 mannitol and cold stress in planta. The labelling into proteins is affected neither when stresses were applied to the plants after incubation with labelled orthophosphate, nor after in vitro protein phosphorylation. This indicates that general protein kinase activities in vivo are strictly limited by P uptake. A marked accumulation of soluble hexoses (mainly sucrose, glucose, and fructose) is observed under imposed stress, suggesting that the inhibition of P uptake in response to hyperosmotic and cold stresses is mediated by sugar accumulation in situ. However, metabolisable sugars like glucose did not alter the entry of phosphate at concentrations of 0.5 mol · L-1, showing that the chemical nature of the osmoticum influences P uptake.

Regulation of Sulfate Assimilation by Nitrogen in Arabidopsis

Using Arabidopsis, we analyzed the effect of omission of a nitrogen source and of the addition of different nitrogen-containing compounds on the extractable activity and the enzyme and mRNA accumulation of adenosine 5′-phosphosulfate reductase (APR). During 72 h without a nitrogen source, the APR activity decreased to 70% and 50% of controls in leaves and roots, respectively, while cysteine (Cys) and glutathione contents were not affected. Northern and western analysis revealed that the decrease of APR activity was correlated with decreased mRNA and enzyme levels. The reduced APR activity in roots could be fully restored within 24 h by the addition of 4 mM each of NO3 −, NH4 +, or glutamine (Gln), or 1 mM O-acetylserine (OAS). 35SO4 2− feeding showed that after addition of NH4 +, Gln, or OAS to nitrogen-starved plants, incorporation of 35S into proteins significantly increased in roots; however, glutathione and Cys labeling was higher only with Gln and OAS or with OAS alone, respectively. OAS strongly increased mRNA levels of all three APR isoforms in roots and also those of sulfite reductase, Cys synthase, and serine acetyltransferase. Our data demonstrate that sulfate reduction is regulated by nitrogen nutrition at the transcriptional level and that OAS plays a major role in this regulation.

Estimating the uptake of traffic-derived NO2 from 15N abundance in Norway spruce needles

The 15N ratio of nitrogen oxides (NOx) emitted from vehicles, measured in the air adjacent to a highway in the Swiss Middle Land, was very high [δ15N(NO2) = +5.7‰]. This high 15N abundance was used to estimate long-term NO2 dry deposition into a forest ecosystem by measuring δ15N in the needles and the soil of potted and autochthonous spruce trees [Picea abies (L.) Karst] exposed to NO2 in a transect orthogonal to the highway. δ15N in the current-year needles of potted trees was 2.0‰ higher than that of the control after 4 months of exposure close to the highway, suggesting a 25% contribution to the N-nutrition of these needles. Needle fall into the pots was prevented by grids placed above the soil, while the continuous decomposition of needle litter below the autochthonous trees over previous years has increased δ15N values in the soil, resulting in parallel gradients of δ15N in soil and needles with distance from the highway. Estimates of NO2 uptake into needles obtained from the δ15N data were significantly correlated with the inputs calculated with a shoot gas exchange model based on a parameterisation widely used in deposition modelling. Therefore, we provide an indication of estimated N inputs to forest ecosystems via dry deposition of NO2 at the receptor level under field conditions.

Phytoextraction of heavy metals from mine soils using hyperaccumulator plants.

Phytoextraction is an environmental-friendly and cost-effective technology that uses metal hyperaccumulator plants to remove heavy metals from soils. The metals are absorbed by the roots, transported and accumulated in the aerial parts of the plants, which can be harvested and eliminated. The aim of this work was to study some hyperaccumulator species that could be useful to decontaminate mine soils and also to investigate the bioavailability and uptake of these metals by plants with the addition of organic amendments. Pot experiments were performed with soil samples collected from two mining areas in the north of Madrid, where there was an intense mining activity more than 50 years ago. Three species (Thlaspi arvense, Brassica juncea and Atriplex halimus) were grown under controlled conditions in pots filled with contaminated soils mixed with 0 Mg, 30 Mg and 60 Mg per hectare of two different organic amendments: a commercial compost made of pine bark, peat and wood fiber and other made of horse and sheep manure and wood fiber. Plants were harvested at the end of their crop cycle and were digested in order to measure metal concentration (Zn, Cu and Cd) in roots and shoots. Highest plant metal concentration was observed in pots treated with pine bark amendment and with pure soil due to an increase in metal bioavailability with decreasing pH. Also in those treatments the total plant biomass was lower, even some plants could not germinate. On the contrary, there was a lower metal concentration in plant tissues of pots with manure because its higher pH whereas plant growth was significantly larger so there was an incresing amount of metals removed from soil by plants. Comparing the three species results indicate a higher total metal uptake in A. halimus than B. juncea and T. arvense. In conclusion, results show that pH affects metal bioavailability and uptake by hyperaccumulator plants. Addition of organic amendments could be a successful technique for stabilization of metals in contaminated soils.

Phytoextraction of heavy metals from mine soils using hyperaccumulator plants

The aim of this work was to study some hyperaccumulator species that could be useful to decontaminate mine soils and also to investigate the bioavailability and uptake of these metals by plants with the addition of organic amendments. Pot experiments were performed with soil samples collected from two mining areas in the north of Madrid, where there was an intense mining activity more than 50 years ago. Three species (Thlaspi arvense, Brassica juncea and Atriplex halimus) were grown under controlled conditions in pots filled with contaminated soils mixed with 0 Mg, 30 Mg and 60 Mg per hectare of two different organic amendments: a commercial compost made of pine bark, peat and wood fiber and other made of horse and sheep manure and wood fiber. Plants were harvested at the end of their crop cycle and were digested in order to measure metal concentration (Zn, Cu and Cd) in roots and shoots. Highest plant metal concentration was observed in pots treated with pine bark amendment and with pure soil due to an increase in metal bioavailability with decreasing pH. Also in those treatments the total plant biomass was lower, even some plants could not germinate. On the contrary, there was a lower metal concentration in plant tissues of pots with manure because its higher pH whereas plant growth was significantly larger so there was an incresing amount of metals removed from soil by plants. Comparing the three species results indicate a higher total metal uptake in A. halimus than B. juncea and T. arvense. In conclusion, results show that pH affects metal bioavailability and uptake by hyperaccumulator plants. Addition of organic amendments could be a successful technique for stabilization of metals in contaminated soils.

Contributions to uncertainty reduction in the estimation of PV plants performance

Esta tesis doctoral presenta un procedimiento integral de control de calidad en centrales fotovoltaicas, que comprende desde la fase inicial de estimación de las expectativas de producción hasta la vigilancia del funcionamiento de la instalación una vez en operación, y que permite reducir la incertidumbre asociada su comportamiento y aumentar su fiabilidad a largo plazo, optimizando su funcionamiento. La coyuntura de la tecnología fotovoltaica ha evolucionado enormemente en los últimos años, haciendo que las centrales fotovoltaicas sean capaces de producir energía a unos precios totalmente competitivos en relación con otras fuentes de energía. Esto hace que aumente la exigencia sobre el funcionamiento y la fiabilidad de estas instalaciones. Para cumplir con dicha exigencia, es necesaria la adecuación de los procedimientos de control de calidad aplicados, así como el desarrollo de nuevos métodos que deriven en un conocimiento más completo del estado de las centrales, y que permitan mantener la vigilancia sobre las mismas a lo largo del tiempo. Además, los ajustados márgenes de explotación actuales requieren que durante la fase de diseño se disponga de métodos de estimación de la producción que comporten la menor incertidumbre posible. La propuesta de control de calidad presentada en este trabajo parte de protocolos anteriores orientados a la fase de puesta en marcha de una instalación fotovoltaica, y las complementa con métodos aplicables a la fase de operación, prestando especial atención a los principales problemas que aparecen en las centrales a lo largo de su vida útil (puntos calientes, impacto de la suciedad, envejecimiento…). Además, incorpora un protocolo de vigilancia y análisis del funcionamiento de las instalaciones a partir de sus datos de monitorización, que incluye desde la comprobación de la validez de los propios datos registrados hasta la detección y el diagnóstico de fallos, y que permite un conocimiento automatizado y detallado de las plantas. Dicho procedimiento está orientado a facilitar las tareas de operación y mantenimiento, de manera que se garantice una alta disponibilidad de funcionamiento de la instalación. De vuelta a la fase inicial de cálculo de las expectativas de producción, se utilizan los datos registrados en las centrales para llevar a cabo una mejora de los métodos de estimación de la radiación, que es la componente que más incertidumbre añade al proceso de modelado. El desarrollo y la aplicación de este procedimiento de control de calidad se han llevado a cabo en 39 grandes centrales fotovoltaicas, que totalizan una potencia de 250 MW, distribuidas por varios países de Europa y América Latina. ABSTRACT This thesis presents a comprehensive quality control procedure to be applied in photovoltaic plants, which covers from the initial phase of energy production estimation to the monitoring of the installation performance, once it is in operation. This protocol allows reducing the uncertainty associated to the photovoltaic plants behaviour and increases their long term reliability, therefore optimizing their performance. The situation of photovoltaic technology has drastically evolved in recent years, making photovoltaic plants capable of producing energy at fully competitive prices, in relation to other energy sources. This fact increases the requirements on the performance and reliability of these facilities. To meet this demand, it is necessary to adapt the quality control procedures and to develop new methods able to provide a more complete knowledge of the state of health of the plants, and able to maintain surveillance on them over time. In addition, the current meagre margins in which these installations operate require procedures capable of estimating energy production with the lower possible uncertainty during the design phase. The quality control procedure presented in this work starts from previous protocols oriented to the commissioning phase of a photovoltaic system, and complete them with procedures for the operation phase, paying particular attention to the major problems that arise in photovoltaic plants during their lifetime (hot spots, dust impact, ageing...). It also incorporates a protocol to control and analyse the installation performance directly from its monitoring data, which comprises from checking the validity of the recorded data itself to the detection and diagnosis of failures, and which allows an automated and detailed knowledge of the PV plant performance that can be oriented to facilitate the operation and maintenance of the installation, so as to ensure a high operation availability of the system. Back to the initial stage of calculating production expectations, the data recorded in the photovoltaic plants is used to improved methods for estimating the incident irradiation, which is the component that adds more uncertainty to the modelling process. The development and implementation of the presented quality control procedure has been carried out in 39 large photovoltaic plants, with a total power of 250 MW, located in different European and Latin-American countries.

The specific features of methionine biosynthesis and metabolism in plants

Plants, unlike other higher eukaryotes, possess all the necessary enzymatic equipment for de novo synthesis of methionine, an amino acid that supports additional roles than simply serving as a building block for protein synthesis. This is because methionine is the immediate precursor of S-adenosylmethionine (AdoMet), which plays numerous roles of being the major methyl-group donor in transmethylation reactions and an intermediate in the biosynthesis of polyamines and of the phytohormone ethylene. In addition, AdoMet has regulatory function in plants behaving as an allosteric activator of threonine synthase. Among the AdoMet-dependent reactions occurring in plants, methylation of cytosine residues in DNA has raised recent interest because impediment of this function alters plant morphology and induces homeotic alterations in flower organs. Also, AdoMet metabolism seems somehow implicated in plant growth via an as yet fully understood link with plant-growth hormones such as cytokinins and auxin and in plant pathogen interactions. Because of this central role in cellular metabolism, a precise knowledge of the biosynthetic pathways that are responsible for homeostatic regulation of methionine and AdoMet in plants has practical implications, particularly in herbicide design.

Galactose-extended glycans of antibodies produced by transgenic plants

Plant-specific N-glycosylation can represent an important limitation for the use of recombinant glycoproteins of mammalian origin produced by transgenic plants. Comparison of plant and mammalian N-glycan biosynthesis indicates that β1,4-galactosyltransferase is the most important enzyme that is missing for conversion of typical plant N-glycans into mammalian-like N-glycans. Here, the stable expression of human β1,4-galactosyltransferase in tobacco plants is described. Proteins isolated from transgenic tobacco plants expressing the mammalian enzyme bear N-glycans, of which about 15% exhibit terminal β1,4-galactose residues in addition to the specific plant N-glycan epitopes. The results indicate that the human enzyme is fully functional and localizes correctly in the Golgi apparatus. Despite the fact that through the modified glycosylation machinery numerous proteins have acquired unusual N-glycans with terminal β1,4-galactose residues, no obvious changes in the physiology of the transgenic plants are observed, and the feature is inheritable. The crossing of a tobacco plant expressing human β1,4-galactosyltransferase with a plant expressing the heavy and light chains of a mouse antibody results in the expression of a plantibody that exhibits partially galactosylated N-glycans (30%), which is approximately as abundant as when the same antibody is produced by hybridoma cells. These results are a major step in the in planta engineering of the N-glycosylation of recombinant antibodies.

Studies of American plants. by Paul C. Standley.

v.22:no.3(1940)

Cost of landspreading and hauling sludge from municipal wastewater treatment plants : case studies.

Mode of access: Internet.

Application of selected industrial engineering techniques to wastewater treatment plants,

"EPA-R2-73-176."

Compilation of reported noble gas effluent monitor capabilities at operating nuclear power plants /

"NUREG-0592."