Made available in DSpace on 2018-08-01T22:33:10Z (GMT). No. of bitstreams: 1 tese_9008_Vitor Heringer.pdf: 1038279 bytes, checksum: 64e5aa0ed12c68d26d3b0b8108d94b8d (MD5) Previous issue date: 2016-07-29 / A seca é um dos fenômenos climáticos que mais causam prejuízos na agricultura influenciando de forma negativa o desenvolvimento socioeconômico. Ela ocorre quando a precipitação apresenta valores abaixo da normal climatológica para determinada região. O diagnóstico da seca pode ser feito por meio de índices quantificadores de seca e análises estatísticas com base em um serie climatológica. Os índices de seca identificam os períodos de seca ou umidade em uma área pontual ou regional a partir de equações empíricas e permitem determinar a intensidade, duração e frequência que ocorrem. Eles relacionam vários anos de variáveis climatológicas como: precipitação, temperatura do ar, evapotranspiração, escoamento superficial e umidade do solo. O objetivo deste trabalho foi avaliar a relação do Índice de Severidade de Seca de Palmer (PDSI) com a produtividade de café no Espírito Santo (ES) e da soja nos estados do Mato Grosso (MT), Paraná (PR) e Rio Grande do Sul (RS). Os cálculos foram realizados por meio do código adaptado de Jacobi et al. (2013), em que são necessários evapotranspiração, precipitação e capacidade de campo. Os dados de produtividade utilizados foram do IBGE, a série histórica de dados utilizada foi no período de 1990 a 2013. O PDSI foi calculado para o ES, RS, PR e MT. Após foi analisado a relação da produtividade com o índice de Palmer. No ES o PDSI foi avaliado com a produtividade de café, no PR, MT e RS o PDSI foi avaliado com a produtividade de soja. De acordo com a regressão linear simples conclui-se que o PDSI não foi significativo na correlação com a produtividade de café no ES, para a cultura da soja o PDSI foi significativo para o estado do RS e MT.

La production de semences très vigoureuses est un levier clépour augmenter les rendements agricoles. Chez le soja, lesrisques de production de semences de faible vigueur germinativesont élevés sous les climats tropicaux, ce qui conduit àune mauvaise installation de la culture, voire un déclassementdes lots en grain. La longévité (c-a-d la capacité de survivre àl’état sec pendant de longues périodes) représente une caractéristiqueimportante de la vigueur germinative. Cependant,la façon dont elle est acquise au cours de la maturation dela graine n’est pas connue. L’objectif de la thèse était d’obtenirun aperçu des mécanismes de régulation de l’acquisitionde la longévité chez le soja. Celle-ci est acquise peu aprèsla phase de remplissage et l’acquisition de la tolérance à ladessiccation (stade R7.2) et continue d’augmenter jusqu’enfi n de maturation. Des différences de longévité ont été observéesentre les récoltes 2013 et 2014.Elles seraient liées auxtempératures élevées au cours de la formation des graines,un facteur connu pour impacter la qualité des semences. Leséquençage Illumina de graines récoltées à 7 stades de développementa révélé des changements complexes se produisantau niveau transcriptionnel dès le stade R7.2, pourtantconsidéré comme correspondant à la maturité physiologique.Les analyses d’enrichissement GO ont révélé une associationpositive entre longévité et les processus de maturation desARN et régulateurs de la post-transcription et une associationnégative avec les processus liés à la photosynthèse. L’acquisi / Soybean is one of the most important oil crop species usedfor food and feed as well as a range of industrial applications.However, producing highly vigorous seeds is a key leverto increase crop production effi ciency. Low physiologicalseed quality, which is more prone to occur under tropicalenvironment, leads to poor stand establishment and decreasedin yields. Seed longevity is the ability to survive the dry statefor prolonged periods of time and represents an importanttrait for seed quality. Here, the objective was to obtain insightsinto the mechanisms regulating the progressive acquisitionof longevity. Using Illumina high-throughput sequencing,RNA was sequenced from seven different stages duringthe acquisition of longevity, generating between 14 and 38million of reads.These reads were aligned to the Glycine maxWm82.a2.v1 gene model. Differentially expressed transcripts(DET) were correlated with the increase in seed longevity.Transcriptome and GO enrichment analyses of these DETrevealed a signifi cant over-representation of terms associatedwith response to stress and RNA processing and modifi cation.Photosynthesis biological process was related to low seedlongevity. HSF and several TF associated with biotic defense(WRKY3 and NLFX1) are candidate genes whose putative rolein seed longevity deserve further characterization. We alsoperformed the determination of the content of non-reducingsoluble sugars, and we observed that the accumulation ofnon-reducing soluble sugars are related to acquisition oflongevity but only the a

Memoria para optar al Título Profesional de: Ingeniera Agrónoma, Mención: Fruticultura / La industria de las hortalizas mínimamente procesadas en fresco (MPF) utiliza comúnmente hipoclorito de sodio (NaClO) para sanitizar tanto el producto como los equipos que se manipulan, sin embargo, existe motivación por reemplazarlo, debido a la generación de subproductos dañinos para la salud. Con este fin, se estudiaron tres sanitizantes alternativos: dióxido de cloro (5 y 10 mg∙L -1 ; OC-5 y OC-10), clorito de sodio acidificado (250 y 500 mg∙L -1 ; CS-250 y CS-500) y ácido peroxiacético (50 y 90 mg∙L -1 ; AP-50 y AP-90), junto con hipoclorito de sodio (100 mg∙L -1 ; HS-100) en germinados de soya (Glycine max) envasados en atmósfera modificada activa (AMA) durante 11 días a 5 ºC. En el primer ensayo se utilizó una bolsa plástica altamente permeable, la cual fue inicialmente barrida con N 2 gaseoso hasta lograr una concentración de 8% O 2 . Los tratamientos HS-100 y CS (250 y 500 mg∙L -1 ) presentaron las mayores reducciones microbianas iniciales (entre 0,7 y 1,8 log ufc∙g -1 ), sin embargo, al final de la conservación todos los recuentos se concentraron alrededor de 8 log ufc∙g -1 para aerobios mesófilos, psicrótrofos y enterobacterias. Sensorialmente, OC-5 obtuvo las mayores puntuaciones tanto en apariencia como intensidad de color hasta el día 7, en contraste, desde el día 7, OC-10 y CS-500 fueron los tratamientos peor evaluados por el panel. Para el segundo ensayo, se escogieron los sanitizantes más efectivos para controlar el crecimiento microbiano, seleccionando de esta forma hipoclorito de sodio (HS-100) dióxido de cloro (OC-5 y OC-10) y clorito de sodio acidificado (CS-250 y CS-500). Se utilizó una bolsa plástica prácticamente impermeable, la cual fue inicialmente barrida con O 2 hasta lograr una concentración de 90% O 2 , alcanzando el día 11 una concentración promedio de 34% O 2 y 49% CO 2 . CS-500 presentó la mayor reducción microbiana inicial (entre 0,8 y 1,1 log ufc∙g -1 ), sin embargo, al final de la conservación todos los valores se concentraron alrededor de 9 log ufc∙g -1 para aerobios mesófilos, psicrótrofos y enterobacterias. La mayor tasa respiratoria la presentó HS-100, que se ajustó también a un mayor gasto de O 2 dentro de la bolsa, pero que no se evidenció en un mayor consumo de azúcar. Sensorialmente, el panel evaluador no encontró diferencias entre tratamientos, que concuerda con la poca variación observada en los parámetros de L, C* y H ab . En ambos ensayos, tanto el contenido de fenoles totales como la actividad antioxidante total no se vieron afectados por la utilización de diferentes sanitizantes. / The industry of minimally processed vegetables commonly uses sodium hypochlorite (NaClO) to sanitize both the product and the equipment to be handled, however, there is interest to replace it due the generation of by-products harmful to health. For this aim, three alternative sanitizers were studied: chlorine dioxide (5 and 10 mg∙L -1 ; OC-5 and OC-10), acidified sodium chlorite (250 and 500 mg∙L -1 ; CS-250 and CS-500) and peroxyacetic acid (50 and 90 mg∙L -1 ; AP-50 and AP-90 ), and as control, sodium hypochlorite (100 mg∙L -1 ; HS-100), in soybean sprouts (Glycine max) under active modified atmosphere packaging for 11 days at 5 °C. In the first assay was used highly permeable plastic bag, which was initially swept with N 2 until a concentration of 8% O 2 . HS-100 and CS (250 and 500 mg∙L -1 ) treatments showed the highest initial microbial reductions (between 0,7 and 1,8 log cfu∙g -1 ), nevertheless, at the end of storage all counts were concentrated around 8 log cfu∙g -1 for aerobic mesophilic, psychrotrophic and enterobacteria. In sensorial evaluation, OC-5 obtained the highest scores in both appearance and color intensity until day 7, in contrast, from day 7, OC-10 and CS-500 were the worst treatments evaluated by the panel. For the second assay, the most effective sanitizers by controlling microbial growth were chosen, thereby, sodium hypochlorite (HS-100), chlorine dioxide (OC-5 and OC-10) and acidified sodium chlorite (CS-250 and CS-500) were selected. The most impermeable plastic bag was used, which was initially swept with O 2 to achieve a concentration of 90% O 2 , reaching for day 11 an average concentration of 34% O 2 and 49% CO 2 . CS-500 showed the highest initial microbial reduction (between 0,8 and 1,1 log cfu∙g -1 ), however, at the end of storage all counts were concentrated around 9 log cfu∙g -1 for aerobic mesophilic, psychrotrophic and enterobacteria. HS-100 displayed the highest respiratory rate, which also adjusted to the highest consumption of O 2 inside the bag, but was not associated with increased consumption of sugar. In sensorial evaluation, panel found no difference between treatments, consistent with low variation observed in the parameters of L, C* and H ab . In both assays, the total phenolic content and total antioxidant activity were not affected by the use of different sanitizers.

Hypoxia as a stressor has physiological implications that have been a focal point for many physiological studies in recent years. In some studies, hypoxia had large effects on the organ tissue degeneration, which ultimately effects multiple ecological processes. These organ tissue studies played a part in the development of new fields like neurocardiology, a specialty that studied the relationship between the brain and the heart. This thesis focuses on how hypoxia-induced cardiac arrest alters the amounts of GLYT2, a glycine reuptake transporter, in the central nervous system of zebrafish, Danio rerio. At 7 days post-fertilization (dpf), zebrafish were exposed to acute, severe hypoxia until they lost equilibrium, and minutes later, subsequent cardiac arrest occurred. Zebrafish were then placed into recovery groups to measure the GLYT2 levels at multiple points in zebrafish recovery. Fish were then sacrificed, and their brains dissected. Using immunofluorescence, the outer left optic tectum of the zebrafish was imaged, and mean image pixel fluorescent intensity was taken. There were significant changes (one-way ANOVA) in the levels of GLYT2 compared to that of the control groups during the course of recovery. GLYT2 levels continued to rise through the 24-hour recovery mark but did not show significant difference after 3 hours of recovery. This suggest that GLYT2 levels increased rapidly in the first 3 hours of recovery and continued to increase through 24 hours at a slower rate. Changes in GLYT2 levels may affect motor and sensory information, movement, visualization, and audition in these zebrafish. Further research should be conducted to determine how long it takes for GLYT2 levels to return to baseline, as well as behavioral measurements through each recovery period as it relates to glycine function.

This document only includes an excerpt of the corresponding thesis or dissertation. To request a digital scan of the full text, please contact the Ruth Lilly Medical Library's Interlibrary Loan Department (rlmlill@iu.edu).

The secretory pathway of G. max performs a prominent role in its defense to H. glycines parasitism, a finding reinforced by the identification of the SNARE-associated gene ALPHA-SNAP-5 being rhg1. Other proteins performing important roles in secretion are Sec61-ALPHA, Sec61-BETA and Sec61-GAMMA which bind to form a trimeric complex that imports proteins into the ER for their eventual secretion. Comparative analyses to the Saccharomyces cerevisiae Sec61ALPHA, Sec61BETA and Sec61GAMMA proteins, respectively, shows G. max has 4 Sec61-ALPHA, 9 Sec61-BETA and4 Sec61-GAMMA proteins. At least one paralog from each gene family is expressed in H. glycines-parasitized root cells in G. max, but while undergoing a defense process. The overexpression of one selected Sec61-ALPHA, Sec61-BETA and Sec61-GAMMA in the H. glycines-susceptible genotype G. max[Williams 82/PI 518671] leads to an engineered defense response. In contrast, RNAi of the same selected Sec61-ALPHA, Sec61-BETA and Sec61-GAMMA genes in the H. glycines-resistant genotype G. max[Peking/PI 548402] leads to an engineered susceptible response. The combined opposite outcomes of the Sec61-ALPHA, Sec61-BETA and Sec61-GAMMA gene overexpression and RNAi provides evidence that they function in the defense process, consistent with the hypothesis that the G. max secretion system plays a role in its defense to H. glycines parasitism.

No description available.

The compatible solute glycine betaine accumulates in many plants including spinach (Spinacea oleracea) under conditions of water deficit stress. The precursor to glycine betaine is choline, a ubiquitous metabolite in plants as a component of phosphotidylcholine. In spinach choline is synthesized from phosphocholine, a product of three sequential N-methylations of phosphoethanolamine catalysed by the cytosolic enzyme S-adenosyl-L-methionine: phosphoethanolamine-N-methyltransferase (PEAMT). PEAMT activity shows diurnal changes with peak activity at the end of the photoperiod and a decrease overnight. The activity of this enzyme is up-regulated 2 to 3-fold in salt-stressed plants relative to unstressed plants. The objective of this thesis is to determine how PEAMT activity is regulated in vivo. Thus, PEAMT activity, protein and transcript levels were quantified in spinach leaves from plants subjected to different light and salinity conditions. A spinach PEAMT eDNA sequence was used to over-express recombinant PEAMT in the protein expression vector pET30a (+). The presence of a polyhistidine-tag on the overexpressed protein allowed for purification by a cobalt metal affinity column. The affinity purified protein was used to produce polyclonal antibodies for immunoblot hybridization analysis. For these studies, PEAMT protein was first immunoaffinity purified from soluble extracts prepared from leaves and then the protein subjected to electrophoresis by SDS-p AGE. Enzyme assays and immunoblot analysis show PEAMT activity and protein levels increase and become relatively constant in leaves of plants exposed to continuous light. In continuous darkness, PEAMT activity and protein levels decrease and remain low and constant. Thus the pattern of changes in PEAMT activity levels are associated with changes in PEAMT protein levels. In contrast, Northern blot hybridizations show that under conditions of constant light, peamt transcript levels undergo cyclical changes with peak levels at 20 and 40 h and troughs at 28 and 52 h after the continuous light treatment was imposed. These peaks coincide with the dark and light cycles of the normal photoperiod. The same cyclical changes in peamt transcript levels was seen for plants transferred from a normal photoperiod to continuous darkness. Since these changes persist in the absence of a day/night cue we conclude that peamt transcript levels are circadian-regulated. The peamt transcript levels of control unstressed and salt-stressed plants also show circadian rhythms, however the levels found in salt-stressed plants were 0.5 to 2-fold higher than the controls. Therefore, while salinization of plants increases peamt transcript abundance, it does not alter the circadian rhythm that transcripts of this gene display. Changes in PEAMT activity and protein levels are likely controlled by other as yet unknown post-translational mechanisms, processes that override and obscure operation of a circadian rhythm in regulating the level of peamt transcripts. / Thesis / Master of Science (MS)

The period gene determines biological rhythmicity in Drosophila melanogaster. The X-linked gene is 7.4kb, containing 8 exons and 7 introns from which a 4.5kb message is translated. A striking feature of the protein encoded by per is a series of alternating threonine-glycine residues in the fifth exon. Moreover, this string of residues is polymorphic for length variation in natural populations, the most frequent variants having 17, 20 or 23 Thr-Gly pairs. In the present study, a geographic analysis of this polymorphism within North American populations was conducted, the results of which indicate significant variation of allele frequency with latitude. The use of spatial autocorrelation analysis and Mantel tests clearly show that the most common variant, encoding 17 Thr-Gly pairs, exhibits a clinal pattern in its distribution along a north-south axis. Furthermore, DNA sequence analysis of several variants has uncovered a novel new variant which encodes 22 Thr-Gly pairs whose nucleotide sequence differs from any published data. Similar statistical analysis conducted on seven allozymes for populations collected along the same transect shows that several have monotonic clinal patterns in their allele frequency distributions which also show correlation with latitude. A previous study of morphological traits on the same populations showed the existence of a non-monotonic clinal pattern. Comparison of the results observed for the molecular and morphological markers indicates that they are subject to different evolutionary forces. The results highlight the importance of comparing patterns of geographic variation using different genetic elements. / Thesis / Master of Science (MS)

NMDA glutamate receptor is a ligand-gated ion channel that mediates a major component of excitatory neurotransmission in the central nervous system (CNS). NMDA receptors are activated by simultaneous binding of two different agonists, glutamate and glycine/ D-serine1. With aging, glutamate concentration gets altered, giving rise to glutamate toxicity that contributes to age-related pathologies like Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, and dementia88,95. Some treatments for these conditions include NMDA receptor blockers like memantine130. However, when completely blocking the receptors, there is a restriction of the receptor's normal physiological function59. A different approach to regulate NMDAR receptors is thorough allosteric modulators that could allow cell type or circuit-specific modulation, due to widely distributed GluN2 expression, without global NMDAR overactivation59,65,122. In one study, we hypothesized that the compound CNS4 selectively modulates NMDA diheteromeric receptors (GluN2A, GluN2B, GuN2C, and GluN2C) based on (three) different glutamate concentrations. Electrophysiological recordings carried out on recombinant NMDA receptors expressed in xenopus oocytes revealed that 30μM and 100μM of CNS4 potentiated ionic currents for the GluN2C and GluN2D subunits with 0.3μM Glu/100μM Gly. However, when using 300μM Glu/100μM Gly, CNS4 inhibited the relative response in the GluN2D subunit and had no effect on the remaining subunits. CNS4 reduced the response to glutamate alone for GluN2A but increased it for GluN2B and did not appear to replace glutamate. Another set of electrophysiological recordings measuring current-voltage relationship was made in order to understand ion flow across the channel in the presence of CNS4. 100μM CNS4 numerically increased the ionic inward current through the channel pore with more positive membrane potential, reflected by a significant difference in reversal potential values, in the GluN2C and GluN2D subunits. CNS4 also exhibited a non-voltage dependent activity and it did not appear to compete with magnesium which naturally blocks the receptor. Finally, the effect of CNS4 on calcium uptake and cellular viability was study in neurons from primary rat brain culture. Cortial and striatal neurons were given excessive doses of synthetic agonist NMDA in order to hyperactivate native NMDAR. In the calcium assay, 100µM of CNS4 significantly increased calcium upatake when given with 300µM NMDA compared with NMDA alone in cortex and when given with 100µM and 300µM NMDA in striatum. In the MTS assay, CNS4 did not alter neuronal viability in either cortical or striatal neurons compared with NMDA alone. Also, when CNS4 was used in non treated neurons it did not alter neuronal viability. Findings from the primary brain culture let us conclude that CNS4 could facilitate calcium influx and possibly be non toxic for neurons. / Master of Science / NMDA ionotropic glutamate receptors are predominately expressed in the central nervous system (CNS). These receptors are activated by glutamate and glycine/ D-serine1. With aging, glutamate concentration in the synapse gets altered giving rise to toxic environments for neurons that can contribute to age-related pathologies like Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, and dementia88,95. Some treatments of these conditions include the receptor blockers like memantine130. However, when completely blocking the receptors, there is a restriction of the receptor's normal physiological function59. A different approach to regulate NMDAR is through allosteric modulators that are compounds that modulate the receptor function without competing with endogenous agonists59,65,122. In this study, we hypothesized that the compound CNS4 selectively modulates NMDAR based on glutamate concentration. Electrophysiological recordings on stage four xenopus oocytes helped us to identify the dose-dependent activity of CNS4 and we found that 30 and 100μM of CNS4 selectively potentiates ionic currents for GluN2C and GluN2D subunits with 0.3μM Glu/100μM Gly but inhibited currents for only GluN2D with 300μM Glu/100μM Gly. Following this, a current-voltage plot was made to examine the channel activity of CNS4. We found a numerical increase of ionic inward current through the channel pore with more positive membrane potential values in the GluN2C and GluN2D subunits. Also, the effect of CNS4 on the ion current activity changed based on glutamate concentration, and CNS4 did not exhibit a voltage-dependent activity, which is a positive feature for compounds that target the receptor133. Finally, to better understand the effect of the compound CNS4 in primary neurons in a toxic environment, a rat brain neuronal culture was made. Increasing doses of NMDA with constant 100µM CNS4 increased cellular Ca2+ influx in a dose-dependent manner. Particularly, 100µM CNS4 with 300µM NMDA exhibited a significant increase in Ca2+ influx in both cortical and striatal neurons compared with 300µM NMDA alone. However, when used alone, 100µM CNS4 did not have an effect on the amount of Ca2+ influx. In addition, CNS4 plus NMDA did not increase viability compared to NMDA alone, and CNS4 alone did not proportionally reduce neuronal viability.