El objetivo de éste estudio fue optimizar un método por Electroforesis Capilar de Zona para la determinación rápida de glutatión reducido (GSH) y oxidado (GSSG) en la matriz de tejido vegetal de hojas y raíces de plantas de tomate (Lycopersicon esculentum L.). Se compararon dos tratamientos de muestra diferentes para el aislamiento de los analitos de la matriz vegetal, se determinó los niveles de GSH y GSSG en plantas sometidas a estrés por cobre en dos periodos de tiempo (48 horas y 28 días) y se evaluó el efecto de cobre sobre la biomasa y los niveles de macronutrientes presentes en la planta. De los resultados obtenidos se concluye que: La optimización de los parámetros instrumentales, tales como largo de capilar, voltaje aplicado y polaridad de la fuente de poder, así como la optimización de los parámetros relacionados con el electrolito de corrida, tales como composición, concentración, pH y fuerza iónica permiten la separación de ambos analitos en un tiempo inferior a los tres minutos. Las condiciones óptimas fueron: polaridad de la fuente positiva, capilar de 40 cm de largo, voltaje de 20 kV, electrolito borato 300 mM, pH 7,6 y 30 segundos de inyección hidrostática de la muestra. El método optimizado para la matriz vegetal presenta una muy buena correlación entre área bajo la curva y concentración de GSH y GSSG, con coeficientes de correlación de 0,9995 y 0,9994, respectivamente. Los límites de detección fueron 2,75 y 1,68 µmol/L para GSH y GSSG y la recuperación del analito en el proceso fue de 105 y 87 %, respectivamente. En la etapa de tratamiento de la muestra, el uso de un método químico (ácido metafosfórico) es más efectivo que el uso de un método físico (copas de ultrafiltración) para el aislamiento de los analitos desde la matriz proteica. Esta efectividad se reflejo en un menor consumo de tiempo, mayor volumen de muestra obtenido y obtención de una matriz mas limpia. El tratamiento prolongado de las plantas con cobre se manifiesta en síntomas de toxicidad con una drástica disminución de la biomasa de la parte aérea y raíz, en asociación con un descenso de la concentración de potasio en ambos órganos. Este efecto no se observa en plantas tratadas por un periodo corto de tiempo con el metal. Los niveles de cobre alcanzados en la parte aérea y raíz correlacionan positivamente con la concentración de cobre aplicada en ambos tratamientos, siendo las concentraciones más altas observadas en la raíz El cobre aplicado a la planta modifica los niveles de glutatión reducido y oxidado tanto en el tratamiento crónico como en el agudo. El efecto más notorio se manifestó sobre el glutatión reducido contenido en la raíz. Este péptido decrece su concentración a medida que incrementa la concentración de cobre, siendo este efecto independiente de los cambios morfológicos que presenta la raíz y del lapso de tiempo en que el cobre fue aplicado. Los cambios producidos en los niveles de glutatión reducido y oxidado constituyen un buen indicador de la respuesta temprana al estrés por metal en plantas desarrolladas, expuestas a cobre por un periodo de tiempo corto (horas)

Cuticular waxes cover all above-ground growing parts of plants. They provide the outermost contact zone between plants and their environment and play a pivotal role in limiting transpirational water loss across the plant surface. The complex mechanisms in cuticular wax biosynthesis conferring proper barrier function still remain to be elucidated. The present study focuses on biosynthetic pathways in wax formation, cuticular wax accumulation and composition and its impact on the epidermal barrier property of the intact system of the astomatous tomato fruit (Lycopersicon esculentum Mill.). Fruits of all developmental stages of the wild type cultivar MicroTom and its lecer6 mutant defective in a β-ketoacyl-CoA synthase involved in very-long-chain fatty acid elongation were analyzed. This 'reverse genetic' approach clarified the importance of the β-ketoacyl-CoA synthase LeCER6 for epidermal barrier property in vivo on the biochemical-analytical level, on the transcriptional level and, furthermore, on the physiological level comparatively between MicroTom wild type and MicroTom lecer6. Surfaces of MicroTom wild type and MicroTom lecer6 fruits showed similar patterns of quantitative wax accumulation, but differed considerably in the permeance for water. Qualitative analyses of the chemical composition of fruit cuticular waxes in the course of fruit development revealed the meaning of the β-ketoacyl-CoA synthase deficiency in the lecer6 mutant. Fruits of this mutant exhibited a distinct decrease in the proportion of n-alkanes of chain lengths > C28. Moreover, a concomitant increase in pentacyclic triterpenoids became discernible in the mature green fruit stage of the mutant. Since quantitative changes of the cutin matrix were not sufficient to affect transpiration barrier properties of the lecer6 mutant presumably the shift in cuticular wax biosynthesis of the lecer6 mutant is responsible for the observed increase of water permeance. In order to investigate the molecular basis of wax formation, a microarray experiment was established that allows the simultaneous and comprehensive analysis of the timing and abundance of transcriptional changes in MicroTom wild type and MicroTom lecer6. This microarray consists of 167 oligonucleotides corresponding to EST and gene sequences of tomato potentially participating in wax biosynthesis, wax modification, transport processes and stress responsiveness. These parameters were correlated with the course of fruit development. This comparison of gene expression patterns showed a variety of differential expressed transcripts encoding for example lipid transfer proteins and the dehydrin TAS14. On the basis of these findings, it can be proposed that diverse regulatory mechanisms like lipid transfer processes or osmotic stress response are affected by the LeCER6 deficiency, which is primarily accompanied by an impaired water barrier property of the fruit cuticle. This present study correlates the continuous increase of LeCer6 gene expression and the accumulation of very-long-chain n-alkanes within the cuticular waxes during the transition from the immature green to the early breaker fruit phase displaying a developmental regulation of the cuticular wax biosynthesis. Organ-specific wax biosynthesis resulted in different cuticular wax pattern in tomato fruits and leaves. Moreover, in contrast to the fruits, LeCER6-deficient leaves showed a significantly reduced wax accumulation, mainly due to a decrease of n-alkanes with chain lengths > C30, while the proportion of pentacyclic triterpenoids were not affected. Deduced from these biochemical-analytical data on tomato fruits and leaves LeCER6 was characterized as a key enzyme in VLCFA biosynthetic pathway responsible for cuticular wax accumulation. In silico analysis of the LeCER6 sequence revealed the presence of two putative transmembrane domains in the N-terminal position. In addition, highly conserved configurations of catalytic residues in the active site of the enzyme were observed, which are probably essential to its overall structure and function in the fatty acid elongation process. High sequence homology of LeCER6 to the very-long-chain condensing enzymes GhCER6 of Gossypium hirsutum L. and AtCER6 of Arabidopsis thaliana (L.) Heynh. was found, which might be a good evidence for similar biochemical functions. Apart from developmental regulation of the cuticular wax biosynthesis, environmental factors influenced the cuticular wax coverage of tomato fruits. Mechanical removal of epicuticular fruit wax evoked large-scale modifications of the quantitative and qualitative wax composition, such as a reduction of aliphatic wax components, and therewith affected the cuticular water permeability. A subsequent regeneration event was included in the regular wax biosynthesis process and led to the compensation of the detached wax amounts and increased the water barrier properties of the cuticular membrane again. In contrast, water-limited conditions had only minor impact on alterations in cuticular wax biosynthesis and, consequently, on the permeance for water of tomato fruits. Floral organ fusion and conditional sterility, as observed in this study, are caused as pleiotropic effects in cell-cell signaling by the loss-of-function mutation in LeCER6. These findings corroborated the functional impact of LeCER6 on the epidermal integrity and are consistent with the current knowledge on eceriferum mutants of Arabidopsis. Investigations of phenotypic and biochemical characteristics of tomato fruits allowed a broader system-orientated perspective of the fruit development of MicroTom wild type and its lecer6 mutant. These analyses highlight more precisely alterations in the fruit surface area, fresh and dry weight, epidermal cell density, photosynthetic activity or glucose content in the course of fruit development. The differences between MicroTom wild type and MicroTom lecer6 characterize very well the large-scale consequences of the LeCER6 deficiency on the physiological status of tomato fruits. Moreover, the results clearly show a part of the genetic controlled network that governs tomato fruit metabolism and mediates extensive changes of the tomato fruit life cycle. The analyses of the stem scar tissue of the tomato fruit revealed a complex set of responses caused by the harvesting process in detail. Throughout storage of the tomato fruits barrier properties were attributed to the suberized stem scar tissue in regard to water loss limitation and reduction of the fungal infection rate. Thereby the endogenous level of abscisic acid was found to be involved in the molecular signaling pathway that regulates the de novo formation of this tissue. For the first time, the chemical composition and physiological importance could be correlated with molecular changes at the transcriptional level during suberization of the stem scar of tomato fruits. In conclusion, this work indicates a novel intact model system for an integrative functional approach for plant barrier properties that was successfully established and carefully studied. The results highlight correlations between wax biosynthesis, distribution of cuticular waxes, and its relevance on the transpirational water loss across the plant surface and, thus, promote the global understanding of plant cuticle biology. / Kutikuläre Wachse bedecken alle oberirdischen Pflanzenteile und stellen somit die Kontaktzone zwischen Pflanzen und ihrer Umwelt dar. Zudem spielen sie eine entscheidende Rolle für den Schutz der Pflanzen vor unkontrolliertem Wasserverlust. Die komplexen Mechanismen der Wachsbiosynthese, die zur dieser Barrierefunktion beitragen, sind jedoch noch weitgehend unaufgeklärt. Die vorliegende Arbeit untersucht Biosynthesewege von kutikulären Wachsen, ihre chemische Beschaffenheit sowie deren funktionelle Bedeutung als Transpirationsbarriere an dem intakten System der astomatären Tomatenfrucht (Lycopersicon esculentum Mill.). Untersuchungen wurden dabei an Früchten unterschiedlicher Entwicklungsstadien des Tomatenkultivars MicroTom Wildtyp und dessen lecer6 Mutante durchgeführt. Die lecer6 Mutante ist durch einen genetisch determinierten Defekt in der β-Ketoacyl-CoA Synthase LeCER6 unfähig zur Verlängerung von sehr langkettigen Fettsäuren. Durch diesen 'reverse genetic' Ansatz wurde der Einfluss der β-Ketoacyl-CoA Synthase LeCER6 auf die Barrierefunktion der Epidermis zunächst in vivo auf der biochemisch-analytischen und physiologischen Ebene vergleichend zwischen MicroTom Wildtyp und MicroTom lecer6 analysiert. Daran schlossen sich Untersuchungen auf transkriptioneller Ebene an. Die den Früchten von MicroTom Wildtyp und der lecer6 Mutante aufgelagerten Wachse unterscheiden sich quantitativ nur wenig, weisen hingegen deutliche Unterschiede in der qualitativen Zusammensetzung und den Wasserleitwerten auf. Die Analyse der chemischen Zusammensetzung der kutikulären Wachse zeigte im Verlauf der Fruchtentwicklung, dass die Defizienz in der β-Ketoacyl-CoA Synthase LeCER6 eine Abnahme des n-Alkananteils in den Wachsen ab einer Kettenlängen > C28 bewirkt, was bereits im Stadium der reifen grünen Früchte zu erkennen ist. Die in der lecer6 Mutante vermehrt eingelagerten pentazyklischen Triterpenoide können die Transpirationsbarriereeigenschaft der aliphatischen n-Alkane nicht adäquat ersetzen. Ein möglicher Einfluss der ebenso untersuchten Kutinmatrix der Tomatenfrucht konnte ausgeschlossen werden. Für eine umfangreiche Genexpressionsanalyse von MicroTom Wildtyp und MicroTom lecer6 wurde ein microarray Experiment konzipiert, welches 167 Oligonukleotide umfasst entsprechend zu bekannten EST- und Gensequenzen der Tomate, die möglicherweise an der Wachsbiosynthese, Wachsmodifikation, relevanten Transportprozessen oder Stressreaktionen beteiligt sind. Der Vergleich der Genexpression zwischen Wildtyp und der lecer6 Mutante zeigte eine Vielzahl von differentiell expremierten Transkripten unter anderem Lipidtransferproteine und das Dehydrin TAS14. Anhand derer kann davon ausgegangen werden, dass der Verlust der LeCER6 Funktion unterschiedliche regulative Mechanismen beeinflusst, wie zum Beispiel Lipidtransportprozesse und Reaktionen des osmotischen Stresses, die mit einer Schwächung der kutikulären Transpirationsbarriere der Fruchtepidermis einhergehen. Die vorliegende Studie belegt zudem erstmals einen Zusammenhang zwischen der Steigerung der LeCer6 Genexpression, der nur geringfügig zeitverzögerten Anreicherung sehr langkettiger n-Alkane in den kutikulären Wachsen und der daraus resultierenden Barriereleistungsfähigkeit. Ebenso wird eine Regulation der kutikulären Wachsbiosynthese in Abhängigkeit von den jeweiligen Stadien der Fruchtentwicklung veranschaulicht. Der organspezifische Vergleich der kutikulären Wachsbiosynthese zeigte, dass sich die Wachsmuster von Früchten und Blättern der Tomatenpflanzen deutlich voneinander unterscheiden. Die Wachsakkumulation auf der Blätterepidermis ist durch die LeCER6-Defizienz hauptsächlich im Anteil sehr langkettiger n-Alkane > C30 signifikant herabgesenkt, während der Gehalt an pentazyklischen Triterpenoiden jedoch nicht, so wie in den Früchten der lecer6 Mutante beobachtet, ansteigt. Aufgrund dieser Untersuchungen der Tomatenfrüchte und -blätter konnte LeCER6 als ein Schlüsselenzym für die Verlängerung sehr langkettiger Fettsäurederivate innerhalb der kutikulären Wachsbiosynthese funktionell charakterisiert werden. Anhand von vergleichenden in silico Sequenzanalysen mit den Fettsäurenelongasen GhCER6 aus Gossypium hirsutum L. und AtCER6 aus Arabidopsis thaliana (L.) Heynh. konnten sowohl zwei mögliche transmembrane Proteindomänen im N-terminalen Bereich als auch hochkonservierte Bereiche im katalytischen Zentrum des LeCER6-Enzyms lokalisiert werden, die vermutlich zur funktionellen Struktur des Enzyms beitragen. Neben der bereits angeführten entwicklungsabhängigen Regulation der Wachsbiosynthese beeinflussen auch Umweltstressoren die kutikuläre Wachsauflage der Tomatenfrüchte. Ein mechanisches Entfernen der epikutikulären Wachse führt zu einer beträchtlichen Reduktion der aliphatischen Wachsbestandteile, welche maßgeblich die Barriereeigenschaft der Kutikulamembran bestimmen. Die einsetzende Regeneration der manipulierten Wachsoberfläche führt zu einer vollständigen Kompensation der entfernten Wachskomponenten, so dass die Tomatenfrüchte in nur kurzer Zeit wieder eine dem Reifestadium entsprechende normale Verteilung der kutikulären Wachse aufweisen. Im Gegensatz dazu führt Wassermangel nur zu sehr geringfügigen qualitativen und quantitativen Veränderungen der kutikulären Wachsschicht und folglich des Wasserleitwertes der Tomatenfrüchte. Die hier dokumentierte Organfusion der Blüte und die eingeschränkte Sterilität der Tomatenpflanzen wurden als pleiotrope Effekte der lecer6 Mutation auf die Zell-Zell-Kommunikation charakterisiert, was der funktionellen Bedeutung von LeCER6 für die Epidermisintegrität entspricht und mit Beobachtungen an eceriferum Mutanten in Arabidopsis übereinstimmt. Die kombinierte Untersuchung phänotypischer und biochemischer Merkmale der Tomatenfrucht erlaubt eine breitere, systemorientierte Gegenüberstellung der Fruchtentwicklung von MicroTom Wildtyp und MicroTom lecer6. Dabei werden durch die Analysen von Größe, Frisch- und Trockengewicht, Dichte der Epidermiszellen, Photosyntheseaktivität und Glukosegehalt der Früchte die Unterschiede zwischen MicroTom Wildtyp und der lecer6 Mutante deutlich aufgezeigt. Die LeCER6-Defizienz der Mutante führt dabei zu weitreichenden Veränderungen im physiologischen Status der Frucht. Diese Ergebnisse spiegeln somit einen Teil des physiologischen Netzwerkes wider, welches weitreichende sekundäre Veränderungen im Lebenszyklus der Tomatenfrucht vermittelt. Das Stielnarbengewebe der Tomatenfrucht wird infolge der Verletzung durch den Ernteprozess gebildet. Basierend auf der de novo Suberinbiosynthese kann diesem Gewebe eine wichtige Barrierefunktion sowohl zur Einschränkung des unkontrollierten Wasserverlustes als auch zur Verringerung der Infektionsrate durch einen pilzlichen Erreger während der Lagerung von Tomatenfrüchten beigemessen werden. Eine Beteiligung der endogenen Abscisinsäure an dem der Bildung des suberinisierten Gewebes der Fruchtstielnarbe zugrunde liegendem, molekularen Signalweg konnte nachgewiesen werden. Zusammenfassend dokumentiert diese Arbeit erstmalig detaillierte Studien im Hinblick auf pflanzliche Barriereeigenschaften an einem intakten Modellsystem. Die präsentierten Ergebnisse zu molekularen Untersuchungen der Wachsbiosynthese und qualitative and quantitative Analysen der Wachsakkumulation werden im Zusammenhang des Schutzes der Pflanzenoberfläche gegen Wasserverlust durch Transpiration diskutiert und bieten somit neue Erkenntnisse über die pflanzliche Kutikula.

A Agricultura Orgânica no Brasil cresce a taxas superiores a 30 % ao ano, devido, principalmente, a uma maior conscientização dos consumidores, que buscam hábitos alimentares mais saudáveis. Dentre as hortaliças, o tomate orgânico representa uma boa oportunidade de investimento para o produtor. No entanto, não existem recomendações técnicas ou genótipos desenvolvidos exclusivamente para esse sistema de produção, o que dificulta a expansão da área sob manejo orgânico. Diante disso, o presente trabalho teve como objetivo avaliar o desempenho de 11 cultivares de tomate (Avalon, Colibri, HTX - 5415, HTX - 8027, Sahel, San Marzano, San Vito, Júpiter, Raminho, Saladete DRW 3410 e Jane) sob dois sistemas orgânicos distintos em ambiente protegido, de março a outubro de 2004, nos municípios de Ibiúna, SP (experimento I) e Araraquara, SP (experimento II). Os experimentos foram conduzidos em propriedades que produzem tomate orgânico em escala comercial. No experimento I, as plantas foram conduzidas em linhas simples, com uma haste e com poda apical drástica, realizada acima do terceiro racemo. As estufas eram conjugadas, e a irrigação realizada por gotejo, utilizado também para a fertiirrigação com biofertilizantes. O espaçamento adotado foi de 1,5 m entre linhas e 0,35 m entre plantas. No experimento II, as estufas não eram conjugadas, e as plantas foram conduzidas em fileiras duplas, com duas hastes por planta, sem poda apical, no espaçamento de 0,8 m entre linhas e 0,6 m entre plantas. Em ambos os experimentos a densidade de plantio foi de 20 mil plantas por hectare. O delineamento experimental adotado foi em blocos ao acaso, com cinco repetições e dez tratamentos (Avalon, Colbri, HTX - 5415, HTX - 8027, Sahel, San Marzano, San Vito, Júpiter, Raminho e Saladete DRW 3410), para o experimento I, e quatro repetições e oito tratamentos (Avalon, Colbri, HTX - 5415, HTX - 8027, Sahel, San Marzano, San Vito, e Jane) para o experimento II. Considerando as condições em que os experimentos foram desenvolvidos, os resultados permitiram concluir que o híbrido Sahel mostrou o melhor desempenho para rendimento comercial, e para outros componentes da produção nos dois sistemas de condução ao qual foi submetido. No experimento I, destacaram-se, ainda, os híbridos Colibri e Saladete DRW 3410, que não diferiram estatisticamente de ‘Sahel’. Os híbridos, de um modo geral, mostraram melhor desempenho que os cultivares de polinização livre, porém, apenas "Sahel" mostrou maior amplitude adaptativa. No experimento I, conduzido com poda apical drástica, a porcentagem de frutos de melhor cotação comercial foi maior do que no experimento II, sem poda apical. O experimento I, também apresentou período de colheita menor e mais concentrado do que no experimento II. A traça-do-tomateiro revelou-se como fator limitante afetando significativamente o rendimento comercial. / Organic agriculture in Brazil has increased about 30% over the last few years since consumers are seeking for healthier foods, i.e. nutritious and free of pesticide residues. Among vegetable crops organically-grown, tomato constitutes an attractive economic opportunity for growers. However, the lack of information about management practices and cultivars adapted to organic production systems under protected cultivation are pointed out as important constraints that prevents this activity to expand. This work aimed to evaluate the performance of 11 indeterminate tomato cultivars (Avalon, Colibri, HTX - 5415, HTX - 8027, Sahel, San Marzano, San Vito, Júpiter, Raminho Sel. IPA, Saladete DRW 3410 and Jane) under two different organic management systems in unheated plastic greenhouse. Two experiments were carried out under climatically different areas of the State of São Paulo (experiment I, Ibiúna county, and experiment II in Araraquara county), from March to October 2004, in farms that commercially produce organic tomatoes under unheated plastic greenhouses. In both experiments tomato plants were staked. Tomato plants in experiment I were conducted with a single stem and apical bud removed upper 3rd cluster. The experiment was fertigated using with biofertilizers. Plant spacing was 1.5 m between rows and 0.35 m between plants. In experiment II, tomato plants were conducted with two stems without apical pruning. They were planted in double line and spaced 0.8 m between lines and 0.6 m between plants. In both experiments plant density was about 20,000 plants ha-1. Experiments were set up in a randomized complete block design. Experiment I had five replications and ten treatments, and experiment II four replications and eight treatments. Sahel hybrid exhibited an outstanding performance for marketable yield in both organic systems but, in experiment I did not differ statistically from hybrids Colibri and Saladete DRW 3410. Hybrid cultivars, in general, outperformed open pollinated cultivars for the evaluated parameters. Apical pruning had significant effect on the increase of tomato fruits graded as of better commercial acceptance (50-60 mm) and also was associated with concentration of the picking period. It was found that tomato pinworm (Tuta absoluta) constitutes a serious limiting pest for organically-grown tomatoes in both regions where the experiments were set up.

Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2018-03-06T18:10:51Z No. of bitstreams: 1 ANDRESSA_VON_WURMB_HELRIGUEL_LEITE_DIS.pdf: 1584575 bytes, checksum: 5f9a44ec42089e160df54cc84eaadb9a (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2018-03-07T14:04:49Z (GMT) No. of bitstreams: 1 ANDRESSA_VON_WURMB_HELRIGUEL_LEITE_DIS.pdf: 1584575 bytes, checksum: 5f9a44ec42089e160df54cc84eaadb9a (MD5) / Made available in DSpace on 2018-03-07T14:11:58Z (GMT). No. of bitstreams: 1 ANDRESSA_VON_WURMB_HELRIGUEL_LEITE_DIS.pdf: 1584575 bytes, checksum: 5f9a44ec42089e160df54cc84eaadb9a (MD5) Previous issue date: 2017-08-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Tomato (Solanum lycopersicum) cultures are frequently attacked by many pathogens, leading to yield losses. Among the pathogens, the fungus Alternaria solani (?Early blight? disease) can cause significant yield reductions and decreases the fruit quality. Due to the high susceptibility of tomato to fungal pathogens, the agricultural practices include the intensive use of agrochemicals, which may result in fruits with high levels of residual pesticides and higher production costs. The promotion of plant defense system represents an additional tool for disease control and plant growth management. Some rhizobacteria have been shown the ability to affect plant development and modulate plant defense against pathogens. Among those bacteria, the genus Streptomyces is able to synthesize molecules that directly interfere with growth, through auxin and ACC deaminase production, and indirectly, through antibiotics and siderophores synthesis. The aim of this work was to select Streptomyces (CLV04, CLV05 and CLV07) with the ability to promote seed germination and plant growth, as well as improve tomato plant resistance against A. solani. Results showed that CLV07 delayed the initial seed germination, but it did not impair the final germination process. However, CLV04, CLV05 and CLV07 reduced the early root and shoot growth. All these rhizobacteria produced high auxin levels. CLV07 was efficient for inducing plant resistance against A. solani. Furthermore, all CLVs showed antagonism by competition, decreasing the A. solani growth. Taking together all results, Streptomyces tested were deleterious for seed and initial plant growth, but CLV07 showed a potential to be used for inducing plant resistance. However, the inoculation of these rhizobacteria on seeds is not recommended. / O tomateiro (Solanum lycopersicum) ? constantemente atacado por diversos pat?genos, levando a perdas de produtividade. Dentre os principais pat?genos, destaca-se o fungo Alternaria solani que causa a doen?a ?pinta-preta? (?Early blight?), capaz de reduzir a produ??o e/ou levar ? forma??o de frutos de baixa qualidade. Devido ? grande susceptibilidade do tomateiro a pat?genos, as pr?ticas agr?colas incluem o uso intensivo de agroqu?micos que resultam, frequentemente, em tomates com altos n?veis residuais de pesticidas. A promo??o do crescimento e da defesa vegetal utilizando microrganismos representa uma ferramenta para o manejo do tomateiro. Dentre as diversas rizobact?rias com capacidade de modular o crescimento e a defesa vegetal, o g?nero Streptomyces vem se destacando. Estas bact?rias s?o capazes de sintetizar mol?culas que interferem tanto diretamente no crescimento, como a auxina e a ACC desaminase, quanto indiretamente, atrav?s da produ??o de antibi?ticos e sider?foros. Pretendeu-se, neste trabalho, selecionar isolados de Streptomyces (CLV04, CLV05 e CLV07) com capacidade de promover a germina??o e o crescimento de plantas de tomate, al?m de aumentar a resist?ncia vegetal contra A. solani. Os resultados demonstram que o CLV07 causou retardo na germina??o, mas n?o reduziu a taxa final de germina??o. Apesar das rizobact?rias CLV04, CLV05 e CLV07 produzirem elevados n?veis de auxina, elas causaram a redu??o do crescimento tanto da raiz quanto da parte a?rea de pl?ntulas de tomate cultivadas in vitro. O CLV07 foi eficiente na promo??o da defesa de plantas de tomate contra A. solani. Todas as rizobact?rias apresentaram antagonismo por competi??o, reduzindo o crescimento do mic?lio do fungo A. solani. Considerando todos os resultados, as diferentes bact?rias Streptomyces foram prejudiciais ? germina??o e ao crescimento inicial de pl?ntulas de tomate. Desta forma, n?o ? recomend?vel a utiliza??o destas bact?rias em sementes e pl?ntulas jovens. Contudo, o CLV07 apresenta potencial para ser utilizado para a indu??o de resist?ncia de plantas de tomate contra o pat?geno A. solani.

Com o objetivo de avaliar novos substratos experimentais e o efeito da aplicação de CO2 via água de irrigação na produção de mudas de tomateiro cultivar 'Hypeel 108', beterraba cultivar 'Early Wonder Tall Top' e repolho cultivar 'Ombrios', cultivadas em bandejas sob ambiente protegido, conduziu-se um experimento na área experimental do Departamento de Produção Vegetal da Escola Superior de Agricultura 'Luiz de Queiroz'/USP, em Piracicaba - SP. O experimento constou de duas etapas: determinação das curvas de fotossíntese em função da radiação fotossinteticamente ativa (PAR) e da concentração de dióxido de carbono para definir o nível de CO2 a ser utilizado na segunda etapa que foi a produção das mudas em substratos experimentais sob dois níveis de CO2 na água de irrigação (presença e ausência de CO2). Para a determinação das curvas utilizou-se o medidor portátil de fotossíntese, modelo LI-6400, sob as concentrações de 200, 365, 500, 600, 800, 900, e 1000mmolCO2 mol-1 e PAR de 700 e 1400mmol-2 s -1. Para a produção de mudas foram utilizados quatro substratos S1, S2, S3 e S4. O delineamento experimental adotado foi em blocos ao acaso, com análise conjunta de dois experimentos. O desenvolvimento das mudas foi avaliado aos 18, 22, 26 e 30 dias à partir da semeadura. Foram avaliadas as variáveis área foliar, massa seca da folha, massa seca do caule e massa seca da raiz e os parâmetros índice de área foliar, taxa de crescimento relativo, taxa de assimilação líquida, taxa de produção de matéria seca e relação parte aérea/sistema radicular. As curvas indicam um resultado favorável das espécies estudadas na concentração de 800mmolCO2mol-1. O substrato S4 foi o que obteve melhores resultados para a produção de mudas de tomateiro, beterraba e repolho, destacando-se por ser o único a apresentar bons resultados nas três espécies avaliadas. Somente as mudas de tomateiro mostraram resultados favoráveis com a aplicação de CO2. / The objective of this study was to evaluate new substrates and the effect of carbon dioxide applied through irrigation water in the production of the following seedlings. These seedling were growth in a controlled environment in an experimental area of the Crop Science Department at ESALQ/USP, Piracicaba, SP. The experiment was divided in two parts. The first part consisted of determining the photosynthesis rate as a function of photosynthesis active radiation (PAR) and carbon dioxide levels to be applied at the second part of the experiment. The second part consisted of seedlings production with 2 levels of CO2 applied through irrigation water. To determine the photosynthesis rate was used a mobile photosynthesis gage (LI-6400), and it wan determined using the following levels of CO2 and PAR: 200, 365, 500, 600, 800, 900 e 1000mmolCO2 mol-1 and 700 and 1400mmol-2 s-1. Four substrates, S1, S2, S3 and S4, were used to grow the seedlings. The experimental design was a randomly blocks with an analyze of the 2 experiments together. The seedlings development were evaluated with 18, 22, 26 and 30 days after planting. The following variable were analyzed: leaf area, leaf dry matter, steam dry matter and root dry matter and the following parameters: leaf area index, growth rate, development rate and the aboveground dry matter/root dry matter rate. The photosynthesis rate showed a good agreement in the plants studies for the 800mmolCO2 mol-1 of CO2. The substrate S4 showed the best result for seedlings production for all crops studied. Only the tomato seedlings showed a good result for CO2application through irrigation water.

Orientador: Hélio Grassi Filho / Banca: Antonio Evaldo Klar / Banca: Rodrigo Domingues Barbosa / Resumo: Devido a escassez da água no planeta, são considerados viáveis estudos que buscam alternativas para economia e melhor aproveitamento da água. Sendo assim a utilização da água de reúso para agricultura e o correto manejo da irrigação torna se uma possibilidade para minimizar o problema da falta de água. Perante isso o objetivo do presente trabalho foi avaliar o efeito de diferentes concentrações e disponibilidades de águas de esgoto doméstica tratado no desenvolvimento e produção do tomate cereja. O cultivo dos tomates foi conduzido na área experimental do Departamento de Engenharia Rural da Faculdade de Ciências Agronômicas da Universidade Estadual Paulista, Campus de Botucatu, no período de agosto a dezembro de 2015. O sistema de irrigação adotado foi localizado por gotejamento, sendo o manejo baseado no tanque classe A. O delineamento experimental foi em sistema fatorial (5x2) com parcelas subdivididas, onde adotou duas lâminas 70 e 100% da evapotranspiração da cultura, a água para os tratamentos foram, água de abastecimento e água de esgoto doméstico tratado com 4 diluições 25, 50, 75 e 100 de reúso, totalizando 10 tratamentos. As avaliações feitas foram: altura; diâmetro do caule; fitomassa verde e seca; teor relativo de água nas folhas (TRA); potencial de água nas folhas (Ψf); teor de nutrientes nas folhas; teor de nutrientes nos frutos; número de frutos maior e menor que 20 mm de diâmetro transversal, número total de frutos; peso total de frutos. Os dados coletados fora... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Due to the scarcity of water on the planet, studies are considered feasible that seek alternatives for economy and better use of water. Therefore, the use of reuse water for agriculture and the correct management of irrigation makes it possible to minimize the problem of lack of water. Therefore, the objective of the present work was to evaluate the effect of different concentrations and availability of treated domestic sewage in the development and production of cherry tomatoes. Tomato cultivation was conducted in the experimental area of the Department of Rural Engineering of the Faculty of Agronomic Sciences of Universidade Estadual Paulista, Campus Botucatu, from August to September 2015. The irrigation system was located by dripping, the management based on the tank class A. The experimental design was in a factorial system (5x2) with subdivided plots, where it adopted two slides 70 and 100% of the crop evapotranspiration, the water for the treatments Were water supply and household sewage treated with 4 dilutions 25, 50, 75 and 100 of reuse, totaling 10 treatments. The evaluations were: height; Stem diameter; Green and dry biomass; Relative leaf water content (TRA); Leaf water potential (Ψf); Nutrient content in leaves; Nutrient content in fruits; Number of fruits larger and smaller than 20 mm of transverse diameter, total number of fruits; Total weight of fruits. The data collected were submitted to analysis of variance and the averages of the tukey test at 5% probability. The results showed that both slides and dilutions of reuse water had significant effects on the agronomic parameters (height, diameter, Ψf, green and dry biomass, nutrient content) during the development of the crop, but for number of fruits, And total weight of fruits, the reuse water had no significant effect, the influence in these parameters was with respect to the slides, plants irrigated with 100% ... / Mestre

As plantas expressam diferentes mecanismos de defesa em resposta a patógenos. Em geral, ocorre o reconhecimento de padrões moleculares associados ao patógeno (PAMPs) por receptores específicos das plantas, desencadeando as respostas de defesa através das vias de sinalização mediadas por jasmonato (JA), etileno (ET) e ácido salicílico (AS). Além disso, a descoberta de sinais endógenos como AtPep1 que regulam a expressão de genes de defesa em Arabidopsis thaliana tem auxiliado na compreensão dos mecanismos de defesa. No entanto, os mecanismos de defesa de plantas a patógenos necrotróficos como Pythium spp. são pouco conhecidos. Desta forma, o desenvolvimento de estratégias para identificar e caracterizar genes de resistência à Pythium spp. pode auxiliar na compreensão dos mecanismos de defesa, possibilitando a obtenção de cultivares resistentes. Uma das estratégias empregadas para a identificação de genes de resistência a patógenos do gênero Pythium consistiu na construção de uma biblioteca de cDNAs diferencialmente expressos em A. thaliana susperexpressando AtPROPEP1, que são mais resistentes à P. irregulare e a P. deliense. Paralelamente, foi realizada a busca de um gene ortólogo ao AtPROPEP1 de A. thaliana em diferentes espécies de solanáceas. Outra estratégia utilizada envolveu a avaliação do efeito da aplicação dos hormônios JA e AS e do regulador de crescimento etefon na resistência de plantas de tomate à P. dissotocum. Posteriormente foi realizado o silenciamento dos genes CTR1, ERF1 e SlPROPEP em plantas de tomate a fim de verificar o papel dos mesmos na resistência das plantas à P. dissotocum. Os resultados obtidos na biblioteca subtrativa indicam que A. thaliana superexpressando AtPROPEP1 pode induzir a expressão não somente da defensina PDF1.2, mas também de GRP3, outro gene envolvido na defesa de plantas contra patógenos. Através da busca de ortólogos de AtPROPEP1 em diferentes solanáceas foi possível identificar um ortólogo em Solanum lycopersicum - SlPROPEP. O tratamento com etefon aumentou a resitência das plantas à P. dissotocum comparado com as plantas não tratadas. Este tratamento também induziu a expressão relativa de SlPROPEP, PR-1, PR-5 e da defensina DEF2. Assim como o tratamento com etefon o silenciamento do gene CTR1, que atua como um regulador negativo da via de sinalização de defesa por ET aumentou a resistência das plantas à P. dissotocum. Enquanto que as plantas silenciadas para os genes ERF1 e SlPROPEP foram mais suscetíveis. O silenciamento de SlPROPEP reprimiu a expressão relativa dos genes PR-1, PR-5, ERF1, LOX-D e DEF2 envolvidos na defesa de plantas à patógenos. Estes resultados, em conjunto com o aumento da resistência nas plantas tratadas com etefon, sugerem que a via de sinalização de ET atua na resistência de tomate à P. dissotocum sendo que esta resistência possivelmente é mediada por SlPROPEP. / Plants express different defense mechanisms in response to pathogens. In general, specific receptors in plants reconize pathogen-associated molecular patterns (PAMPs), triggering the response defenses by signaling pathways mediated by jasmonate (JA), ethylene (ET) and salicylic acid (SA). Furthermore, the discovery of endogenous signals, as AtPep1, which regulate defense genes expression in Arabidopsis thaliana, has aided the understanding of the defense mechanisms. However, the defense mechanisms of plants to necrotrophic pathogens such as Pythium spp. are poorly known. Thus, development of strategies to identify and characterize the resistance genes to Pythium spp. can help the understand of defense mechanisms, allowing the obtainance of resistant cultivars. One of the strategies used for the identification of resistance genes to pathogens within the genus Pythium consisted to construct a cDNA library of differentially expressed genes in plants overexpressing AtPROPEP1, which are more resistant to P. irregulare and P. deliense. Simultaneously, a search for orthologous genes to AtPROPEP1 from A. thaliana in different species of Solanaceae was performed. Another strategy used involved the evaluation of application of JA and AS and of the growth regulator ethephon on the resistance of tomato plants to P. dissotocum. Then, the silencing of the genes CTR1, ERF1 and SlPROPEP was performed in tomato plants in order to check the role of the tomato resistance to P. dissotocum. The results obtained from subtractive cDNA library indicate that A. thaliana plants overexpressing AtPROPEP1 may induce expression not only of the PDF1.2 defensin but also the GRP3 expression, another gene involved in plant defense against pathogens. By the search for orthologous genes to AtPROPEP1 in different species of Solanaceae was possible to identify an ortholog in Solanum lycopersicum - SlPROPEP. The ethephon treatment increased resistance of plants to P. dissotocum compared to untreated plants. This treatment also induced the expression of SlPROPEP, PR-1, PR-5 and DEF2 defensin. As ethephon treatment, the silencing of CTR1 gene, which acts as a negative regulator of defense signaling pathway by ET, increased plant resistance to P. dissotocum. Whereas the plants containing ERF1 and SlPROPEP genes silenced were more susceptible. SlPROPEP silencing repressed the relative expression of the genes PR-1, PR-5, ERF1, LOX-D e DEF2 involved in plant defense to pathogens. These results, together with increased resistance in the plants treated with ethephon, suggest that ET signaling pathway acts in tomato to resistance P. dissotocum and this resistance is likely mediated by SlPROPEP.

Atualmente a tomaticultura brasileira ostenta o título de cultivo com o maior custo de produção médio no país. Dentre algumas alternativas a fim de minimizar os elevados gastos está o uso de híbridos de crescimento semideterminado em sistema de meia-estaca, devido a menor necessidade de podas, insumos e mão de obra. Realizaram-se dois experimentos, em condições de campo aberto, com os objetivos de (a) determinar o efeito de diferentes sistemas de poda (P1, P2 e P3) e distintos espaçamentos entre plantas (E1 e E2) sobre a produção e seus componentes para o híbrido Silvety, de hábito de crescimento semideterminado do segmento Salada Longa Vida (SLV) (Experimento 1); e (b) avaliar o efeito de distintos sistemas de poda (P1 e P2) sobre a produção e seus componentes para os híbridos Silvety, Sagatan e H003, de hábito de crescimento semideterminado do segmento SLV (Experimento 2). Em ambos os experimentos adotou-se o delineamento em blocos ao acaso com os tratamentos em esquema fatorial 3 x 2 e quatro repetições. O experimento 1 foi conduzido em Jacuí, MG, de maio de 2015 a novembro de 2015, já o experimento 2 foi conduzido em Holambra, SP, de fevereiro de 2016 a agosto de 2016. Avaliaram-se o número total de frutos (NTF), a produção total por parcela (PTP), o número de frutos comercializáveis (NFC), a produção comercial (PCO), a produção por planta (PPP), a massa média dos frutos (MMF), o número de frutos por classe AAA, AA e A (NAAA, NAA e NA), a massa de frutos por classe AAA, AA e A (MAAA, MAA e MA), o número de hastes (NH) o número de inflorescências (NI), o número de frutos por inflorescência (NFI), o número de frutos com rachaduras cuticulares (CUT), o número de frutos ocos (OCO) e o número de frutos manchados (MAN). Nas condições em que os experimentos foram conduzidos e de acordo com os resultados obtidos conclui-se que no experimento 1, a poda P2e o espaçamento E1 mostraram-se os mais indicados para o híbrido Silvety. No experimento 2, novamente, a poda P2 mostrou ser a mais recomendada para os híbridos Silvety, Sagatan e H003. / Currently, the Brazilian tomato crop holds the title with the highest average cost of production in the country. Among the alternatives to minimize the high expenses, is the use of hybrids with semi-determinate growth habit in a training system of \"half-stakes\", due to the less need of pruning, inputs and labor. Two experiments were carried out under open field conditions, aiming to (a) to determine the effect of different pruning system (P1, P2 and P3) and spacing between plants (E1 and E2) on the production and its components for the Silvety hybrid, of semi-determinate growth habit of varietal segment Salad Extended Shelf Life (SLV) (Experiment 1), and (b) to evaluate the effect of pruning system (P1 and P2) on the production and its components for the Silvety, Sagatan and H003 hybrids, of semi- determinate growth habit of segment SLV (Experiment 2). In both experiments it was adopted the randomized block design with the treatments in a 3 x 2 factorial scheme, and four replications. Experiment 1 was conducted in Jacuí, MG, from May 2015 to November 2015, and Experiment 2 was conducted in Holambra, SP, from February 2016 to August 2016. There were assessed the total number of fruits (NTF), the total production per plot (PTP), the number of commercial fruits (NFC), the commercial production (PCO), the production per plant (PPP), the average mass of fruits (MMF), the number of fruits per classes AAA,AA and A (NAAA, NAA and NA), the mass of fruits per classes AAA, AA and A (MAAA, MAA and MA), the number of stems (NH), the number of clusters (NI), number of fruits per cluster (NFI), number of fruits with micro-cracking (CUT), number of hollow fruits (OCO), number of fruits with blotchy (MAN). Under the conditions that the trials were conducted and according to the results obtained, it was concluded that in experiment 1, the pruning system P2 and E1 spacing were the most suitable for Silvety hybrid. In experiment 2, again, the pruning system P2 showed to be the most recommended for Silvety, Sagatan and H003 hybrids.

Orientador: Rumy Goto / Banca: Omulo Fujito Kobori / Banca: Roberto Botelho Ferraz Branco / Resumo: Realizou-se dois experimentos, sendo o primeiro com objetivo em avaliar o melhor estádio para se realizar a aplicação foliar da mistura de auxina (Ax), citocinina (CK) e giberelina (GA), em tomate determinado (Solanum lycopersicum L.), utilizando-se a dose de 0,20% do produto. O segundo experimento com objetivo em avaliar o efeito da "poda belga" e a influência da mistura de auxina (Ax), citocinina (CK) e giberelina (GA) no desenvolvimento e metabolismos das plantas. O primeiro experimento foi conduzido na área experimental da Faculdade de Ciências Agronômicas da Universidade Estadual Paulista-UNESP, em São Manuel. Foi utilizado delineamento de blocos ao acaso sendo 16 tratamentos, quatro repetições: T1: Testemunha; T2: Antes do transplante; T3: Antes do transplante + Enraizamento; T4: Antes do transplante + Enraizamento + Desbrota; T5: Antes do transplante + Enraizamento + Desbrota + Início do Florescimento; T6: Antes do transplante + Enraizamento + Desbrota + Início do Florescimento+ Frutificação; T7: Enraizamento; T8: Enraizamento + Desbrota; T9: Enraizamento + Desbrota + Ínicio do Florescimento; T10: Enraizamento + Desbrota + Início do Florescimento+ Frutificação; T11: Desbrota (mesmo dia); T12: Desbrota ... / Abstract: Two experiments were conducted, the first with the aim to evaluate the best stage for performing foliar application of the mixture of auxin (Ax), cytokinins (CK) and gibberellins (GA), in particular tomato (Solanum lycopersicum L.), using a dose of 0,20% of the product. The second experiment in order to evaluate the effect of "Belgian pruning" and the influence of mixing auxin (Ax), cytokinin (CK) and gibberellic acid (GA) in the development and metabolism of plants. The first experiment was conducted in the experimental area of the Faculty of Agricultural Sciences at the Universidade Estadual Paulista-UNESP, in São Manuel. The experimental design was randomized blocks with 16 treatments, four replications: T1:Control; T2: Before Transplant; T3: Before Transplant + Rooting; T4: Before Transplant + Rooting + Sprouting; T5: Before Transplant + Rooting + Sprouting + Starts Flowering; T6: Before Transplant + Rooting + Sprouting + Starts Flowering+ Starts Fruiting; T7: Rooting; T8: Rooting + Sprouting; T9: Rooting + Sprouting + Ínicio do Florescimento; T10: Rooting + Sprouting + Starts Flowering+ Starts Fruiting; T11: Sprouting (mesmo dia); T12: Sprouting + Starts Flowering; T13: Sprouting + Starts Flowering+ Starts Fruiting; T14: Starts Flowering; T15: Starts Flowering+ Starts Fruiting; T16: Starts Fruiting. The second experiment was conducted ... / Mestre

Orientador: Marcos Ferreira Minicucci / Coorientador: Sergio Alberto Rupp de Paiva / Coorientador: Paula Schmidt Azevedo Gaiolla / Banca: Leonardo Antonio Mamede Zornoff / Banca: Helio Vannucchi / Resumo: Introdução: A insuficiência cardíaca é uma frequente complicação após o infarto agudo do miocárido (IAM), sendo que a remodelação cardíaca apresenta papel de destaque nesse cenário. O aumento do estresse oxidativo e do processo inflamatório estão associados com a remodelação cardíaca após o IAM. Devido ao grande impacto sócio econômico e às altas taxas de mortalidade, torna-se relevante a identificação de outros fatores que modulem o processo de remodelação cardíaca e podemos destacar a suplementação de alimentos com propriedades antioxidantes, como o tomate (Lycopersicon esculentum), e um de seus principais constituintes, o licopeno. Objetivo: Avaliar a influência da suplementação de tomate e licopeno na remodelação cardíaca após o IAM. Materiais e métodos: Ratos Wistar machos, com peso entre 200 e 250g, foram submetidos ao infarto experimental ou à cirurgia simulada e alocados em seis grupos: 1) grupo Sham (animais submetidos à cirurgia simulada) alimentado com dieta padrão (grupo- SC); 2) grupo Sham alimentado com dieta suplementada com licopeno 1mg/kg peso corporal/dia (grupo-SL); 3) grupo Sham alimentado com dieta suplementada com tomate (grupo-ST); 4) grupo infartado (animais submetidos ao infarto experimental) alimentado com dieta padrão (grupo-IC); 5) grupo infartado alimentado com dieta suplementada com licopeno 1mg/kg de peso corporal/dia (grupo-IL); 6) grupo infartado alimentado com dieta suplementada com tomate (grupo-IT). O licopeno e o tomate das dietas foram diluídos em óleo de milho para serem adicionados à ração (0,5ml de óleo/kg de peso corporal/dia) e os grupos controles tiveram a mesma quantidade de óleo adicionado. Após três meses de observação os animais foram submetidos ao estudo funcional, morfométrico, e bioquímico. Os valores obtidos foram apresentados em média ± desvio padrão. As variáveis que não preencheram os critérios de normalidade e de... / Abstract: Introduction: Heart failure is a common complication after the acute myocardial infaction (AMI), and cardiac remodeling has a prominent role in this scenario. Increased oxidative stress and inflammation are associated with cardiac remodeling after AMI. Due to the great economic and social and high mortality rates, it's important to identify other factors that modulate the process of cardiac remodeling and among this factors are the supplementation of foods with antioxidant properties, such as tomato (Lycopersicon esculentum), and of its primary constituents, the lycopene. Objectives: Evaluate the influence of tomato and lycopene supplementation in cardiac remodeling after AMI. Methods: Male Wistar rats, weighing between 200 and 250g, were submitted to the experimental infarction or sham surgery and divided into six groups: 1) Sham group (animals undergoing sham surgery) fed a standard diet (SC group); 2) Sham group fed diet supplemented with 1 mg lycopene/kg body/day (SL group); 3) Sham group fed diet supplemented with tomato (ST group); 4) infarcted group (animals submitted to experimental infarction) fed a standard diet (IC group); 5) infarcted group fed diet supplemented with 1 mg lycopene/kg of body weight/day (IL group); 6) infarcted group fed diet supplemented with tomato (IT group). The lycopene and the tomato were dilute in corn oil to be add into the diet (0.5 ml of corn oil/kg body weight/day) and the standard group received the same oil. After three months of observation the animals underwent functional, morphometric and biochemical analysis. The values were expressed as mean ± standard deviation. The variables that did not meet the criteria of normality and equal variance were normalized before analysis. Comparisons between groups were made by two-way analysis of variance (ANOVA). For non-normalized data, we used Kruskal-Wallis test (comparison between the Sham groups, SC, SL and ... / Mestre