Soil aggregation in a crop-livestock integration system under no-tillage

Grazing intensities can influence soil aggregation, which can be temporarily and permanently affected. The objective of this study was to evaluate the aggregate stability in water at the end of a soybean cycle and during pasture development in a crop-livestock integration system under no-tillage and grazing intensities. The experiment was initiated in 2001, in a dystrophic Red Latosol, after soybean harvest. Treatments consisted of pasture (black oat + Italian ryegrass) at heights of 10, 20 and 40 cm, grazed by young cattle, and a control (no grazing), followed by soybean cultivation, in a randomized block design. Soil samples were collected at the end of the soybean cycle (May/2007), during animal grazing (September/2007) and at the end of the grazing cycle (November/2007). The grazing period influences aggregate distribution, since in the September sampling (0-5 cm layer), there was a higher proportion of aggregates > 4.76 mm at all grazing intensities. Soil aggregation is higher in no-tillage crop-livestock integration systems in grazed than in ungrazed areas.

Nutrient relations during an eucalyptus cycle at different population densities

To synchronize nutrient availability with the requirements of eucalyptus during a cultivation cycle, the nutrient flow of this system must be well understood. Essential, for example, is information about nutrient dynamics in eucalyptus plantations throughout a cultivation cycle, as well as impacts on soil nutrient reserves caused by the accumulation and subsequent export of nutrients via biomass. It is also important to quantify the effect of some management practices, such as tree population density (PD) on these fluxes. Some nutrient relations in an experiment with Eucalyptus grandis, grown at different PDs in Santa Barbara, state of Minas Gerais, Brazil, were evaluated for one cultivation cycle. At forest ages of 0.25, 2.5, 4.5, and 6.75 years, evaluations were carried out in the stands at seven different PDs (between 500 and 5,000 trees ha-1) which consisted in chemical analyses of plant tissue sampled from components of the aboveground parts of the tree, from the forest floor and the litterfall. Nutrient contents and allocations of the different biomass components were estimated. In general, there were only small and statistically insignificant effects of PD on the nutrient concentration in trees. With increasing forest age, P, K, Ca and Mg concentrations were reduced in the aboveground components and the forest floor. The magnitud of biochemical nutrient cycling followed the sequence: P > K > N > Mg. At the end of the cycle, the quantities of N, P, Ca and Mg immobilized in the forest floor were higher than in the other components.

Cycle length of spermatogenesis in shrews (mammalia: soricidae) with high and low metabolic rates and different mating systems.

The aim of the present study was to establish and compare the durations of the seminiferous epithelium cycles of the common shrew Sorex araneus, which is characterized by a high metabolic rate and multiple paternity, and the greater white-toothed shrew Crocidura russula, which is characterized by a low metabolic rate and a monogamous mating system. Twelve S. araneus males and fifteen C. russula males were injected intraperitoneally with 5-bromodeoxyuridine, and the testes were collected. For cycle length determinations, we applied the classical method of estimation and linear regression as a new method. With regard to variance, and even with a relatively small sample size, the new method seems to be more precise. In addition, the regression method allows the inference of information for every animal tested, enabling comparisons of different factors with cycle lengths. Our results show that not only increased testis size leads to increased sperm production, but it also reduces the duration of spermatogenesis. The calculated cycle lengths were 8.35 days for S. araneus and 12.12 days for C. russula. The data obtained in the present study provide the basis for future investigations into the effects of metabolic rate and mating systems on the speed of spermatogenesis.

Hydrological processes obtained on the plot scale under four simulated rainfall tests during the cycle of different crop systems

The cropping system influences the interception of water by plants, water storage in depressions on the soil surface, water infiltration into the soil and runoff. The aim of this study was to quantify some hydrological processes under no tillage cropping systems at the edge of a slope, in 2009 and 2010, in a Humic Dystrudept soil, with the following treatments: corn, soybeans, and common beans alone; and intercropped corn and common bean. Treatments consisted of four simulated rainfall tests at different times, with a planned intensity of 64 mm h-1 and 90 min duration. The first test was applied 18 days after sowing, and the others at 39, 75 and 120 days after the first test. Different times of the simulated rainfall and stages of the crop cycle affected soil water content prior to the rain, and the time runoff began and its peak flow and, thus, the surface hydrological processes. The depth of the runoff and the depth of the water intercepted by the crop + soil infiltration + soil surface storage were affected by the crop systems and the rainfall applied at different times. The corn crop was the most effective treatment for controlling runoff, with a water loss ratio of 0.38, equivalent to 75 % of the water loss ratio exhibited by common bean (0.51), the least effective treatment in relation to the others. Total water loss by runoff decreased linearly with an increase in the time that runoff began, regardless of the treatment; however, soil water content on the gravimetric basis increased linearly from the beginning to the end of the rainfall.

MICROBIAL CHARACTERISTICS OF SOILS UNDER AN INTEGRATED CROP-LIVESTOCK SYSTEM

Integrated crop-livestock systems (ICLs) are a viable strategy for the recovery and maintenance of soil characteristics. In the present study, an ICL experiment was conducted by the Instituto Agronômico do Paraná in the municipality of Xambre, Parana (PR), Brazil, to evaluate the effects of various grazing intensities. The objective of the present study was to quantify the levels of microbial biomass carbon (MBC) and soil enzymatic activity in an ICL of soybean (summer) and Brachiaria ruziziensis (winter), with B. ruziziensis subjected to various grazing intensities. Treatments consisted of varying pasture heights and grazing intensities (GI): 10, 20, 30, and 40 cm (GI-10, GI-20, GI-30, and GI-40, respectively) and a no grazing (NG) control. The microbial characteristics analysed were MBC, microbial respiration (MR), metabolic quotient (qCO2), the activities of acid phosphatase, β-glucosidase, arylsuphatase, and cellulase, and fluorescein diacetate (FDA) hydrolysis. Following the second grazing cycle, the GI-20 treatment (20-cm - moderate) grazing intensity) contained the highest MBC concentrations and lowest qCO2 concentrations. Following the second soybean cycle, the treatment with the highest grazing intensity (GI-10) contained the lowest MBC concentration. Soil MBC concentrations in the pasture were favoured by the introduction of animals to the system. High grazing intensity (10-cm pasture height) during the pasture cycle may cause a decrease in soil MBC and have a negative effect on the microbial biomass during the succeeding crop. Of all the enzymes analyzed, only arylsuphatase and cellulase activities were altered by ICL management, with differences between the moderate grazing intensity (GI-20) and no grazing (NG) treatments.

Role of the renal circadian timing system in maintaining water and electrolytes homeostasis.

Many basic physiological functions exhibit circadian rhythmicity. These functional rhythms are driven, in part, by the circadian clock, an ubiquitous molecular mechanism allowing cells and tissues to anticipate regular environmental events and to prepare for them. This mechanism has been shown to play a particularly important role in maintaining stability (homeostasis) of internal conditions. Because the homeostatic equilibrium is continuously challenged by environmental changes, the role of the circadian clock is thought to consist in the anticipative adjustment of homeostatic pathways in relation with the 24h environmental cycle. The kidney is the principal organ responsible for the regulation of the composition and volume of extracellular fluids (ECF). Several major parameters of kidney function, including renal plasma flow (RPF), glomerular filtration rate (GFR) and tubular reabsorption and secretion have been shown to exhibit strong circadian oscillations. Recent evidence suggest that the circadian clock can be involved in generation of these rhythms through external circadian time cues (e.g. humoral factors, activity and body temperature rhythms) or, trough the intrinsic renal circadian clock. Here, we discuss the role of renal circadian mechanisms in maintaining homeostasis of water and three major ions, namely, Na(+), K(+) and Cl(-).

Small RNA-dependent expression of secondary metabolism is controlled by Krebs cycle function in Pseudomonas fluorescens.

Pseudomonas fluorescens CHA0, an antagonist of phytopathogenic fungi in the rhizosphere of crop plants, elaborates and excretes several secondary metabolites with antibiotic properties. Their synthesis depends on three small RNAs (RsmX, RsmY, and RsmZ), whose expression is positively controlled by the GacS-GacA two-component system at high cell population densities. To find regulatory links between primary and secondary metabolism in P. fluorescens and in the related species Pseudomonas aeruginosa, we searched for null mutations that affected central carbon metabolism as well as the expression of rsmY-gfp and rsmZ-gfp reporter constructs but without slowing down the growth rate in rich media. Mutation in the pycAB genes (for pyruvate carboxylase) led to down-regulation of rsmXYZ and secondary metabolism, whereas mutation in fumA (for a fumarase isoenzyme) resulted in up-regulation of the three small RNAs and secondary metabolism in the absence of detectable nutrient limitation. These effects required the GacS sensor kinase but not the accessory sensors RetS and LadS. An analysis of intracellular metabolites in P. fluorescens revealed a strong positive correlation between small RNA expression and the pools of 2-oxoglutarate, succinate, and fumarate. We conclude that Krebs cycle intermediates (already known to control GacA-dependent virulence factors in P. aeruginosa) exert a critical trigger function in secondary metabolism via the expression of GacA-dependent small RNAs.

Role of β-TrCP1, variant and homologue in HIV-1 life cycle

Summary The CD4 molecule plays a key role in AIDS pathogenesis, it is required for entry of the virus into permissive cells and its subsequent down-modulation of the cell surface is a hallmark of HN-1 infected cells. The virus encodes no less than three proteins that participate in this process: Nef, Vpu and Env. Vpu protein interacts with CD4 within the endoplasmic reticulum of infected cells, where it targets CD4 for degradation through the interaction with a cellular protein named ß-TrCP1. This F-box protein functions as the substrate recognition subunit of the SCF ß-Trcr E3 ubiquitin ligase, which normally induce the ubiquitination and subsequent degradation of various proteins such as ß-catenin and IxBa. Mammals possess a homologue of ß-TrCP1, HOS, also named ß-TrCP2 which has a cytoplasmic subcellular distribution. Structural analysis of the ligand-binding domain of both homologues shows striking surface similarities. Both F-box proteins have a redundant role in a number of cellular processes; however the potential role of ß-TrCP2 in HIV-1 infected cells has not been evaluated. In the present study, we assessed the existence of génetic variants of BRTC, encoding ß-TrCP1, and evaluated whether these variants would affect CD4 down-modulation. Additionally, we determined whether ß-TrCP2 shares with its homologue structural and functional properties that would allow it to bind Vpu, modulate CD4 expression, and thus participate in HN-1 pathogenesis. We identified a single nucleotide polymorphism present in the human population with an allelic frequency of 0.03 that leads to the substitution of alanine 507 by a serine. However, we showed by transient transfection in HeLa CD4+ cells that this variant behaves as ß-TrCP1 with respect to CD4 down-modulation. We established transient expression systems in HeLa CD4+ cells to test whether ß-TrCP2 is implicated in Vpu-mediated CD4 down-modulation. We show by coimmunoprecipitation experiments that ß-TrCP2 binds Vpu and is able to induce CD4 down-modulation as efficiently as ß-TrCP1. In two different cell lines, HeLa CD4+ and Jurkat, Vpu-mediated CD4 down-modulation could not be completely reversed through the silencing of endogenous ß-TrCP 1 or ß-TrCP2 individually, but required both genes to be silenced simultaneously. We evaluated the role of ß-TrCP1 and ß-TrCP2 in HIV-1 life cycle using silencing prior to actual viral infection. Both ß-TrCP1 and ß-TrCP2 contributed to CD4 down-modulation during aone-cycle viral infection iri Ghost cells. In addition, the combined silencing of both homologues in the absence of env and nef reversed CD4 down-modulation, showing that ß-TrCP 1 and ß-TrCP2 represent the main and additive effectors of HIV-1 encoded Vpu. In addition, we showed that silencing of ß-TrCPI but not ß-TrCP2 induced a decrease of HIV-1 LTR-driven expression. In a transient transfection system with Tat and a LTR luciferase reporter, both homologues modulated LTR-driven expression. The present study revealed that ß-TrCP2 represents a novel protein participating in HIV-1 cycle and complete comprehension of the complex interplay occurring between the two F-Box will improve our understanding of HIV-1 infection. Résumé La molécule CD4 joue un rôle clef dans la pathogenèse du SIDA ; elle est requise pour l'entrée du virus dans les cellules permissives et la diminution de sa concentration au niveau de la surface cellulaire est une importante caractéristique des cellules infectées par le VIH-1. Le virus encode pas moins de trois protéines qui participent à ce processus Nef, Vpu et Env. La protéine Vpu lie CD4 au niveau du réticulum endoplasmique et induit sa dégradation en interagissant avec une protéine cellulaire nommée ß-TrCP 1. Cette protéine de type F-Box est une sous unité du complexe ubiquitine-ligase E3 SCFß-TrCP. Elle permet la reconnaissance du substrat par le complexe qui induit l'ubiquitination et la subséquente dégradation de diverses protéines cellulaires comme la ß-catenin ou IκBα. Les mammifères possèdent un homologue à ß-TrCP1appelé ß-TrCP2 (ou HOS). L'analyse comparative du domaine permettant la reconnaissance des substrats des deux homologues montre de frappantes similarités. Le rôle de ß-TrCP2 dans le cycle viral du VIH-1 n'a pas encore été évalué. Lors de cette étude, nous avons recherché l'existence de variants génétique de BTRC (codant pour ß-TrCP1) et nous avons évalué si ces variants pourraient affecter la dégradation des molécules CD4 induite par le virus. Nous avons ainsi identifié un polymorphisme présent dans la population humaine avec une fréquence allélique de 0.03 qui consiste en une substitution de l'alanine 507 par une sérine. Nous avons cependant montré par transfection dans des cellules HeLa CD4+ que ce variant se comporte comme ß-TrCP 1 en ce qui concerne la modulation de CD4. De plus, nous avons déterminé si ß-TrCP2 partageait avec son homologue des propriétés structurelles et fonctionnelles qui lui permettraient de lier Vpu, moduler la concentration de CD4 et ainsi prendre part à la pathogenèse du SIDA. Pour ce faire, nous avons établi un système d'expression temporaire dans des cellules HeLa CD4+. Par co-immunoprécipitation, nous avons montré que ß-TrCP2 lie Vpu et est capable d'induire la dégradation de CD4 aussi efficacement que ß-TrCP1. Dans deux différentes lignées cellulaires, HeLa CD4+ et Jurkat, la dégradation de CD4 n'a pu être complètement inhibée par le silencing individuel de ß-TrCP 1 ou ß-TrCP2, mais nécessitait le silencing simultané des 2 gènes. Nous avons évalué le rôle des deux homologues dans le cycle viral du VIH-1 en infectant des cellules Ghost avec le virus après avoir effectué un silencing des deux protéines. Nous avons ainsi montré que ß-TrCP 1 et ß-TrCP2 contribuent de manière additive à la dégradation de CD4 induite par une infection du VIH-1. Le silencing combiné des deux homologues inhiba complètement cette dégradation en l'absence de env et nef, prouvant qu'aucune autre voie ne participe à ce processus: En outre, nous avons montré que le silencing de ß-TrCP 1 mais pas celui de ß-TrCP2 induisait une diminution de l'expression virale sous contrôle du LTR. Nous n'avons cependant pas été en mesure de reconstituer cet effet en exprimant Tat et un gène reporteur sous contrôle du LTR dans des cellules HeLa CD4+. Le présent travail révèle que ß-TrCP2 représente une nouvelle protéine participant dans le cycle viral du VIH-1. Une complète compréhension de l'effet de chacun des deux homologues sur le cycle viral permettra d'améliorer notre compréhension de l'infection par le VIH-1.

Inclusion of soil erosion impacts in life cycle assessment on a global scale: application to energy crops in Spain

Purpose: Despite the fundamental role of ecosystem goods and services in sustaining human activities, there is no harmonized and internationally agreed method for including them in life cycle assessment (LCA). The main goal of this study was to develop a globally applicable and spatially resolved method for assessing land-use impacts on the erosion regulation ecosystem service.Methods: Soil erosion depends much on location. Thus, unlike conventional LCA, the endpoint method was regionalized at the grid-cell level (5 arc-minutes, approximately 10×10 km2) to reflect the spatial conditions of the site. Spatially explicit characterization factors were not further aggregated at broader spatial scales. Results and discussion: Life cycle inventory data of topsoil and topsoil organic carbon (SOC) losses were interpreted at the endpoint level in terms of the ultimate damage to soil resources and ecosystem quality. Human health damages were excluded from the assessment. The method was tested on a case study of five three-year agricultural rotations, two of them with energy crops, grown in several locations in Spain. A large variation in soil and SOC losses was recorded in the inventory step, depending on climatic and edaphic conditions. The importance of using a spatially explicit model and characterization factors is shown in the case study.Conclusions and outlook: The regionalized assessment takes into account the differences in soil erosion-related environmental impacts caused by the great variability of soils. Taking this regionalized framework as the starting point, further research should focus on testing the applicability of the method trough the complete life cycle of a product and on determining an appropriate spatial scale at which to aggregate characterization factors, in order to deal with data gaps on location of processes, especially in the background system. Additional research should also focus on improving reliability of the method by quantifying and, insofar as it is possible, reducing uncertainty.

A system-level, molecular evolutionary analysis of mammalian phototransduction

Visual perception is initiated in the photoreceptor cells of the retina via the phototransduction system.This system has shown marked evolution during mammalian divergence in such complex attributes as activation time and recovery time. We have performed a molecular evolutionary analysis of proteins involved in mammalianphototransduction in order to unravel how the action of natural selection has been distributed throughout thesystem to evolve such traits. We found selective pressures to be non-randomly distributed according to both a simple protein classification scheme and a protein-interaction network representation of the signaling pathway. Proteins which are topologically central in the signaling pathway, such as the G proteins, as well as retinoid cycle chaperones and proteins involved in photoreceptor cell-type determination, were found to be more constrained in their evolution. Proteins peripheral to the pathway, such as ion channels and exchangers, as well as the retinoid cycle enzymes, have experienced a relaxation of selective pressures. Furthermore, signals of positive selection were detected in two genes: the short-wave (blue) opsin (OPN1SW) in hominids and the rod-specific Na+/Ca2+,K+ ion exchanger (SLC24A1) in rodents. The functions of the proteins involved in phototransduction and the topology of the interactions between them have imposed non-random constraints on their evolution. Thus, in shaping or conserving system-level phototransduction traits, natural selection has targeted the underlying proteins in a concerted manner.

Passive immunization with neutralizing antibodies interrupts the mouse mammary tumor virus life cycle.

Mouse mammary tumor virus (MMTV) infects the host via mucosal surfaces and exploits the host immune system for systemic spread and chronic infection. We have tested a neutralizing rat monoclonal antibody specific for the retroviral envelope glycoprotein gp52 for its efficiency in preventing acute and chronic mucosal and systemic infection. The antibody completely inhibits the superantigen response and chronic viral infection following systemic or nasal infection. Surprisingly however, the antibody only partially inhibits the early infection of antigen-presenting cells in the draining lymph node. Despite this initially inefficient protection from infection, superantigen-specific B- and T-cell responses and systemic viral spread are abolished, leading to complete clearance of the retroviral infection and hence interruption of the viral life cycle. In conclusion, systemic neutralizing monoclonal antibodies can provide an efficient protection against chronic retroviral amplification and persistence.

Urea and sugarcane straw nitrogen balance in a soil-sugarcane crop system

The objectives of this study were to evaluate nitrogen utilization by sugarcane ratoon from two sources, applied urea and sugarcane straw covering soil surface (trash blanket), besides the recovery of N from both sources in the soil-plant system. The following treatments were established in a randomized block design with four replicates: T1, vinasse-urea (100 kg ha-1 of urea-N) mixture applied on the total area of the soil covered with cane trash labeled with 15N; T2, vinasse-urea mixture (urea labeled with 15N; 100 kg ha-1 of urea-N) applied on the total area of the soil covered with non-labeled sugarcane trash; and T3, urea-15N (100 kg ha-1 of urea-N) applied in furrows at both sides of cane rows, with previous surface application of vinasse, onto soil without trash covering. The vinasse was applied at a rate of 100 m³ ha-1 in all treatments. The experiment was carried out on a Yellow Red Podzolic soil (Paleudalf), from October 1997 to August 1998, in Piracicaba, SP, Brazil. The nitrogen use efficiency of urea by the sugarcane ratoon was 21%, while that of the sugarcane straw was 9%. The main contributions of N from sugarcane trash, during one cycle, are the preservation and increase of the organic N in soil. The tendency for a lower accumulation of urea-N in the sugarcane plant, in the soil surface covered with sugarcane residue, was compensated by the assimilation of N from trash mineralization. Nitrogen derived from cane trash was more available to plants in the second half of the ratoon cycle

MOLECULAR MECHANISMS OF A ROOT SYSTEM TO SOIL ADAPTATION

Contrairement aux animaux, les plantes sont des organismes sessiles qui ne possèdent pas de mécanismes de fuite quand les conditions environnementales ne sont plus optimales. Les plantes sont physiquement ancrées à l'endroit où elles ont germées et aux conditions environnementales qui parfois peuvent être extrêmes. Les possibilités d'acclimatation de différentes espèces, parfois même de groupes de plantes au sein d'une même espèce, peuvent varier mais repose sur une adaptation génétique de la plante. L'adaptation est un long processus qui repose sur l'apparition spontanée de mutations génétiques, leur mise à l'épreuve face aux conditions environnementales, et dans le cas où la mutation a un impact positif sur la survie dans cet habitat particulier, elle sera maintenue dans une population donnée de plantes. De telles populations, appelées écotypes, sont le matériel de départ pour la découverte de gènes qui induisent un bénéfice pour la plante dans un environnement donné. La plante la plus étudiée en biologie moléculaire est Arabidopsis thaliana, l'arabette des prés. Dans une étude précédente, les racines d'écotypes naturels d'Arabidopsis ont été comparées et un écotype, Uk-1, avait le système racinaire le plus particulier. Cet écotype possède des racines beaucoup plus courtes et plus ramifiées que tous les autres écotypes. Des analyses plus poussées ont montré qu'une seule mutation dans un gène était la cause de ce phénotype, le gène BREVIS RADIX (BRX), mot latin signifiant 'racine courte'. Bien que l'on connaisse le gène BRX, on connaît finalement peu de choses sur son importance adaptative. Dans cette étude, nous avons montré que la mutation dans le gène BRX rend la plante plus résistante aux sols acides. Dans l'optique de mieux comprendre cette valeur adaptative du mutant brx, nous avons analysé dans quels tissus le gène BRX jouait un rôle important. Nous avons pu mettre en évidence que BRX est important pour le développement du protophloème. Le protophloème est un élément du système vasculaire de la plante. En général, les plantes supérieures possèdent deux systèmes de transport à longue distance. L'un d'eux, appelé xylème, transporte l'eau et les nutriments absorbés du sol par les racines vers les feuilles. Les feuilles sont le siège du processus de photosynthèse au cours duquel sont produits des sucres qui devront être distribués partout dans les autres parties de la plante. Le tissu cellulaire chargé de livrer les produits de la photosynthèse, ainsi que les régulateurs de croissance, est le phloème. Ce dernier regroupe le métaphloème et le protophloème. Le protophloème est essentiel pour la livraison des sucres synthétisés ainsi que des signaux de croissance aux pointes des racines, centres organogéniques responsables de la production de nouvelles cellules durant la phase de croissance de la racine. La structure du protophloème peut être décrite comme des tubes continus, vides et résistants, faits de cellules spécialisées qui permettent un transport efficace et rapide. Nous avons montré que dans les mutants brx ces canaux de transports sont discontinus car certaines cellules n'ont pas terminé leur cycle de différenciation. Ces cellules obstruent le conduit ce qui fait que les sucres et les signaux de croissance, comme l'auxine, ne peuvent plus être transportés aux méristèmes. En conséquence, la prolifération de l'activité des méristèmes est compromise, ce qui explique les racines courtes. Au lieu d'être délivré aux méristèmes, l'auxine se concentre en amont des méristèmes où cela provoque l'apparition de nouvelles racines branchées et, très probablement, l'activation des pompes à protons. Sur des sols acides, la concentration en ion H+ est très élevée. Ces ions entrent dans les cellules de la racine par diffusion et perturbent notablement la croissance des racines et de la plante en général. Si les cellules de la racine possédaient des pompes à protons hyperactives, elles seraient capable d'évacuer le surplus d'ions H+ en dehors de la cellule, ce qui leur assurerait de meilleures chances de survie sur sols acides. De fait, le mutant brx est capable d'acidifier le milieu de culture dans lequel il est cultivé plus efficacement que la plante sauvage. Ce mutant est également capable de donner plus de progéniture sur ce type de milieu de croissance que les plantes sauvages. Finalement, nous avons trouvé d'autres mutants brx en milieu naturel poussant sur sols acides, ce qui suggère fortement que la mutation du gène BRX est une des causes de l'adaptation aux sols acides. -- Plants as sessile organisms have developed different mechanisms to cope with the complex environmental conditions in which they live. Adaptation is the process through which traits evolve by natural selection to functionally improve in a given environmental context. An adaptation to the environment is characterized by the genetic changes in the entire populations that have been fixed by natural selection over many generations. BREVIS RADIX (BRX) gene was found through natural Arabidopsis accessions screen and was characterized as a root growth regulator since loss-of-function mutants exhibit arrested post-embryonic primary root growth in addition to a more branched root system. Although brx loss-of-function causes a complete alteration in root architecture, BRX activity is only required in the root vasculature, in particular in protophloem cell file. Protophloem is a part of the phloem transport network and is responsible for delivery of photo-assimilates and growth regulators, coming from the shoot through mature phloem component - metaphloem, to the all plant primary meristems. In order to perform its function, protophloem is the first cell file to differentiate within the root meristem. During this process, protophloem cells undergo a partial programmed cell death, during which they build a thicker cell wall, degrade nucleus and tonoplast while plasma membrane stays functional. Interestingly, protophloem cells enter elongation process only after differentiation into sieve elements is completed. Here we show that brx mutants fail to differentiate protophloem cell file properly, a phenotype that can be distinguished by a presence of a "gap" cells, non-differentiated cells between two flanking differentiated cells. Discontinuity of protophloem differentiation in brx mutants is considered to be a consequence of local hyperactivity of CLAVATA3/EMBRYO SURROUNDING REGION 45 (CLE45) - BARELY ANY MERISTEM 3 (BAM3) signaling module. Interestingly, a CLE45 activity, most probably at the level of receptor binding, can be modulated by apoplastic pH. Altogether, our results imply that the activity of proton pumps, expressed in non-differentiated cells of protophloem, must be maintained under certain threshold, otherwise CLE45-BAM3 signaling pathway will be stimulated and in turn protophloem will not differentiate. Based on vacuolar morphology, a premature cell wall acidification in brx mutants stochastically prevents the protophloem differentiation. Only after protophloem differentiates, proton pumps can be activated in order to acidify apoplast and to support enucleated protophloem multifold elongation driven by surrounding cells growth. Finally, the protophloem differentiation failure would result in an auxin "traffic jam" in the upper parts of the root, created from the phloem-transported auxin that cannot be efficiently delivered to the meristem. Physiologically, auxin "leakage" from the plant vasculature network could have various consequences, since auxin is involved in the regulation of almost every aspect of plant growth and development. Thus, given that auxin stimulates lateral roots initiation and growth, this scenario explains more branched brx root system. Nevertheless, auxin is considered to activate plasma membrane proton pumps. Along with this, it has been shown that brx mutants acidify media much more than the wild type plants do, a trait that was proposed as an adaptive feature of naturally occurring brx null alleles in Arabidopsis populations found on acidic soils. Additionally, in our study we found that most of accessions originally collected from acidic sampling sites exhibit hypersensitivity to CLE45 treatment. This implies that adaptation of plants to acidic soil involves a positive selection pressure against upstream negative regulators of CLE45-BAM3 signaling, such as BRX. Perspective analysis of these accessions would provide more profound understanding of molecular mechanisms underlying plant adaptation to acidic soils. All these results are suggesting that targeting of the factors that affect protophloem differentiation is a good strategy of natural selection to change the root architecture and to develop an adaptation to a certain environment. -- Les plantes comme organismes sessiles ont développé différents mécanismes pour s'adapter aux conditions environnementales complexes dans lesquelles elles vivent. L'adaptation est le processus par lequel des traits vont évoluer via la sélection naturelle vers une amélioration fonctionnelle dans un contexte environnemental donné. Une adaptation à l'environnement est caractérisée par des changements génétiques dans des populations entières qui ont été fixés par la sélection naturelle sur plusieurs générations. Le gène BREVIS RADIX (BRX) a été identifié dans le crible d'une collection d'accessions naturelles d'Arabidopsis et a été caractérisé comme un régulateur de la croissance racinaire étant donné que le mutant perte-de-fonction montre une croissance racinaire primaire arrêtée au stade post-embryonnaire et présente de plus un système racinaire plus ramifié que la plante sauvage. Bien que le mutant perte-de-fonction brx cause une altération complète de l'architecture racinaire, l'activité de BRX n'est requise que dans la vascularisation racinaire, en particulier au niveau du protophloème. Le protophloème est un composant du réseau de transport du phloème et est responsable du transit des dérivés de la photosynthèse ainsi que des régulateurs de croissances, venant de la partie aérienne par le phloème mature (métaphloème) vers tous les méristèmes primaires de la plante. Pour pouvoir réaliser sa fonction, le protophloème est la première file de cellules à se différencier à l'intérieur du méristème de la racine. Pendant ce processus, les cellules du protophloème subissent une mort cellulaire programmée partielle durant laquelle elles épaississent leur paroi cellulaire, dégradent le noyau et le tonoplaste tandis que la membrane plasmique demeure fonctionnelle. De manière intéressante, les cellules du protophloème entament le processus d'allongement seulement après que la différenciation en tubes criblés soit complète. Ce travail montre que le mutant brx est incapable de mener à bien la différenciation de la file de cellules du protophloème, phénotype qui peut être visualisé par la présence de cellules 'trous', de cellules non différenciées entourées de deux cellules différenciées. La discontinuité de la différenciation du phloème dans le mutant brx est considérée comme la conséquence de l'hyperactivité localisée du module de signalisation CLA VA TA3/EMBRYO SURROUNDING REGION 45 (CLE45) - BARELY ANY MERISTEM 3 (BAM3). De manière intéressante, l'activité de CLE45, très probablement au niveau de la liaison avec le récepteur, peut être modulé par le pH apoplastique. Pris ensemble, nos résultats impliquent que l'activité des pompes à protons, actives dans les cellules non différenciées du protophloème, doit être maintenue en dessous d'un certain seuil autrement la cascade de signalisation CLE45-BAM3 serait stimulée, en conséquence de quoi le protophloème ne pourrait se différencier. D'après la morphologie vacuolaire, une acidification prématurée de la paroi cellulaire dans le mutant brx empêche la différenciation du protophloème de manière stochastique. Une fois que le protophloème se différencie, les pompes à protons peuvent alors être activées afin d'acidifier l'apoplaste et ainsi faciliter l'allongement des cellules énuclées du protophloème, entraînées par la croissance des cellules environnantes. Finalement, la différenciation défectueuse du protophloème produit une accumulation d'auxine dans la partie supérieure de la racine car le phloème ne peut plus acheminer efficacement l'auxine au méristème. Physiologiquement, la 'fuite' d'auxine à partir du réseau vasculaire de la plante peut avoir des conséquences variées puisque l'auxine est impliquée dans la régulation de la majorité des aspects de la croissance et développement de la plante. Etant donné que l'auxine stimule l'initiation et développement des racines latérales, ce scénario pourrait expliquer le système racinaire plus ramifié du mutant brx. En plus, l'auxine est considérée comme un activateur des pompes à protons. Par ailleurs, nous avons montré que les mutants brx ont la capacité d'acidifier le milieu plus efficacement que les plantes sauvages, une caractéristique des populations sauvages <¥Arabidopsis poussant sur des sols acides et contenant les allèles délétés brx. De plus, dans nos résultats nous avons mis en évidence que la plupart des accessions collectées originellement sur des sites acidophiles montre une hypersensibilité au traitement par CLE45. Ceci implique que l'adaptation des plantes aux sols acides repose sur la pression de sélection positive à rencontre des régulateurs négatifs de CLE45- BAM3, situés en amont de la cascade, tel le produit du gène BRX. Les analyses de ces accessions pourraient aboutir à une meilleure compréhension des mécanismes moléculaires responsables de l'adaptation des plantes aux sols acides. Tous nos résultats suggèrent que le ciblage des facteurs affectant la différenciation du protophloème serait une stratégie gagnante dans la sélection naturelle pour changer l'architecture de la racine et ainsi s'adapter efficacement à un nouvel environnement.

Planning, Developing, and Implementing the Iowa Pavement Marking Management System (IPMMS): Phases I and II, 2006

With an annual pavement marking program of approximately $2 million and another $750 thousand invested in maintenance of durable markings each year, the Iowa DOT is seeking every opportunity to provide all-year markings staying in acceptable condition under all weather conditions. The goal of this study is to analyze existing pavement marking practices and to develop a prototype Pavement Marking Management System (PMMS). This report documents the first two phases of a three-phase research project. Phase I includes an overview of the Iowa DOT’s existing practices and a literature review regarding pavement marking practices in other states. Based on this information, a work plan was developed for Phases II and III of this study. Phase II organized the key components necessary to develop a prototype PMMS for the Iowa DOT. The two primary components are (1) performance/life cycle curves for pavement marking products, and (2) an application matrix tailored to the pavement marking products and roadway and environmental conditions faced by the Iowa DOT. Both components will continue to be refined and tailored to Iowa materials and conditions as more performance data becomes available.

Automated User Interface testing in Symbian Operating System

Tässä diplomityössä tutkitaan automatisoitua testausta ja käyttöliittymätestauksen tekemistä helpommaksi Symbian-käyttöjärjestelmässä. Työssä esitellään Symbian ja Symbian-sovelluskehityksessä kohdattavia haasteita. Lisäksi kerrotaan testausstrategioista ja -tavoista sekä automatisoidusta testaamisesta. Lopuksi esitetään työkalu, jolla testitapausten luominen toiminnalisuus- ja järjestelmätestaukseen tehdään helpommaksi. Graafiset käyttöliittymättuovat ainutlaatuisia haasteita ohjelmiston testaamiseen. Ne tehdään usein monimutkaisista komponenteista ja niitä suunnitellaan jatkuvasti uusiksi ohjelmistokehityksen aikana. Graafisten käyttöliittymien testaukseen käytetään usein kaappaus- ja toistotyökaluja. Käyttöliittymätestauksen testitapausten suunnittelu ja toteutus vaatii paljon panostusta. Koska graafiset käyttöliittymät muodostavat suuren osan koodista, voitaisiin säästää paljon resursseja tekemällä testitapausten luomisesta helpompaa. Käytännön osuudessa toteutettu projekti pyrkii tähän tekemällä testiskriptien luomisesta visuaalista. Näin ollen itse testien skriptikieltä ei tarvitse ymmärtää ja testien hahmottaminen on myös helpompaa.