Submitted by Jorge Cativo (jorge.cativo@inpa.gov.br) on 2018-08-23T14:38:12Z No. of bitstreams: 2 Dissertação Matheus Miranda Caniato.pdf: 2877850 bytes, checksum: 9f5a34aab328acaeb1c5681aefacab65 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-08-23T14:38:12Z (GMT). No. of bitstreams: 2 Dissertação Matheus Miranda Caniato.pdf: 2877850 bytes, checksum: 9f5a34aab328acaeb1c5681aefacab65 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-01-30 / The use of rhizobacteria to increase crop productivity by promoting growth and management of the target spot through the induction of systemic resistance to plant diseases are shown as tool capable of allowing the expansion of tomato North region. Thus, the objectives of this work were to evaluate the influence of biochar in the population of cultivable soil bacteria, besides the root colonization capacity of bacterial isolates, growth promotion, phosphate solubilization, indole acetic acid production and induction of resistance to Corynespora almost. The influence of the isolates origin on the mentioned variables was also evaluated. As well as the identification of the bacterial isolates. The influence of the bio-carbon on the population of cultivable soil bacteria was carried out using the colony-forming unit counting methodology. Verification of root colonization was performed using techniques for visualizing turbidity zones in tubes with agar-water (0.8%), optical microscopy and root plating. Growth promotion and induction of resistance were evaluated in greenhouse and the experimental design adopted for both experiments was completely randomized and inoculation of the bacteria was performed by microbiolization. Identification was performed by sequencing the 16S rRNA gene. The results indicate that doses up to 34 t ha-1 positively influence the population of cultivable soil bacteria in the 0-10 cm layer. The three methods used to evaluate root colonization were efficient and complement each other in the interpretation of the results. The isolates ISO4T2, ISO22T3, ISO53T1, ISO17T3 and ISO113T1 promoted an increment in the confirmation test and present potential to be evaluated in the field in the future. Isolates 7T1, 114T1 and 52T2 induced partial resistance against C. cassiicola and present potential to be evaluated under field conditions. The origin of the isolates has influence on the ability to colonize the root system of tomato plants and solubilize phosphate, but does not affect the other variables. The partial sequencing of the 16S rRNA gene allowed the identification of 23 isolates at the genus and group level, but not in species. / A inoculação de rizobactérias visando o aumento da produtividade das culturas, por meio da promoção de crescimento e o manejo da mancha alvo por meio da indução de resistência sistêmica a doenças em plantas, mostram-se como ferramentas capazes de permitirem a expansão do cultivo de tomate na Região Norte. Assim, os objetivos desse trabalho foram avaliar a influência do biocarvão sobre a população de bactérias cultiváveis do solo, além da capacidade de colonização radicular dos isolados bacterianos, promoção de crescimento, solubilização de fosfato, produção de ácido indól acético e indução de resistência a Corynespora casiicola. A influência da origem dos isolados sobre as variáveis mencionadas também foi avaliada. Assim como foi realizada a identificação dos isolados bacterianos. A influência do biocarvão sobre a população de bactérias cultiváveis do solo foi realizada por meio da metodologia de contagem de unidade formadora de colônias. A verificação da colonização radicular foi realizada por meio das técnicas de visualização de zonas de turbidez em tubos com ágar-água (0,8%), microscópia de luz e plaqueamento de raízes. A promoção de crescimento e a indução de resistência foram avaliadas em casa de vegetação e o delineamento experimental adotado para ambos os experimentos foi o inteiramente casualizado e a inoculação das bactérias foi realizada por microbiolização. A identificação foi realizada por meio do sequenciamento do gene 16S rRNA. Os resultados indicaram que doses até 34 t ha-1 influenciaram de forma positiva a população de bactérias cultiváveis do solo na camada de 0-10 cm. Os três métodos utilizados para avaliação da colonização radicular se mostraram eficientes e se complementaram na interpretação dos resultados. Os isolados ISO4T2, ISO22T3, ISO53T1, ISO17T3 e ISO113T1 promoveram incremento no teste de confirmação e apresentaram potencial para serem avaliados futuramente em campo. Os isolados, 7T1, 114T1 e 52T2 induziram resistência parcial a C. cassiicola e apresentaram potencial para serem avaliados em condição de campo. A origem dos isolados influenciou a capacidade de colonizar o sistema radicular de plântulas de tomateiro e de solubilizar fosfato. O sequenciamento parcial do gene 16S rRNA, permitiu identificar 23 isolados em nível de gênero e grupo, mas não em espécie.

Submitted by Inácio de Oliveira Lima Neto (inacio.neto@inpa.gov.br) on 2018-08-23T18:02:17Z No. of bitstreams: 1 Dissertação Final PDF.pdf: 14440240 bytes, checksum: 2af4bf3f84a50787393d47d1dd9daac4 (MD5) / Made available in DSpace on 2018-08-23T18:02:17Z (GMT). No. of bitstreams: 1 Dissertação Final PDF.pdf: 14440240 bytes, checksum: 2af4bf3f84a50787393d47d1dd9daac4 (MD5) Previous issue date: 2018-07-31 / The bacterial wilt, caused by Ralstonia solanacearum, is one of the major diseases that affect Tomato cultivation in tropical climates, limiting production. The present work had as objective to evaluate the potential of solanaceous species as rootstock in commercial tomato, on agronomic characteristics, fruit quality and control of bacterial wilt. The experiment was conducted in a greenhouse, in a randomized complete block design, with four blocks, and six treatments, being the rootstocks: Cubiu (Solanum sessiliflorum); two cultivars of Jiló (Solanum aethiopicum), long pale green and large hill; Jurubebão (Solanum lycocarpum) and Jurubeba Juna (Solanum stramonifolium), grafted with the cultivar Santa Cruz Kada, susceptible to the disease. The glue index, plant height and number of leaves at 15 days after grafting, stem diameter, graft stem diameter and graft point diameter at 30, 60 , 90 and 120 days after transplanting. The fruits were collected regularly for the evaluation of the total number of fruits, number of marketable fruits, number of scrap fruits, average mass of marketable fruits and scrap, vertical diameter and horizontal diameter, yield of marketable fruits, . Fruit quality was measured by the analysis of total soluble solids, pulp pH, titratable acidity and soluble solids ratio and titratable acidity. The incidence of the disease was evaluated weekly after the first wilting plant was investigated. Morphological and histological observations of the healing callus of the evaluated treatments were performed. All evaluated rootstocks presented glue index ≥ 93.33%, indicating that there was no initial incompatibility between the combinations evaluated. The number of commercial fruits differed significantly in the rootstocks, which were long, light green and covered, being the treatment with lower total number of fruits and number of commercial fruits among the evaluated treatments. Cubiu presented the lowest values of commercial fruit production and productivity in relation to treatments, except for the jurubeba juna in which it did not differ. There was no incidence of bacterial wilt in the long-green jiló and jurubeba juna rootstocks, which conferred disease resistance on the grafted plants. The histological sections showed incompatibility characteristics of the tomato evaluated with the cubiu, jurubebão and jurubeba juna rootstocks. The rootstocks, long jiló light green and jurubeba juna presented high index of adhesion, resistance to bacterial wilt, and productivity statistically equal to the control in the conditions of the present work, so that they have great potential for use as rootstocks in the crop of the tomato. Key-words: Solanaceae, Ralstonia solanacearum, Solanum lycopersicum, Resistance. / A murcha-bacteriana, causada por Ralstonia solanacearum, é uma das principais doenças que acometem o cultivo do Tomateiro em climas tropicais, limitando a produção. O presente trabalho teve como objetivo avaliar a potencialidade de espécies de solanáceas como porta-enxerto em tomate comercial, sobre características agronômicas, qualidade dos frutos e no controle da murcha bacteriana. O experimento foi conduzido em casa de vegetação, em delineamento experimental em blocos ao acaso, com quatro blocos, e seis tratamentos sendo eles os porta-enxertos: Cubiu (Solanum sessiliflorum); duas cultivares de Jiló (Solanum aethiopicum), a comprido verde claro e a morro grande; Jurubebão (Solanum lycocarpum) e Jurubeba Juna (Solanum stramonifolium), enxertados com o tomateiro cultivar Santa Cruz Kada, susceptível à doença. Foram avaliados o índice de pegamento, a altura da planta e o número de folhas aos 15 dias após a enxertia, e o diâmetro do caule do porta-enxerto, o diâmetro do caule do enxerto e o diâmetro do ponto de enxertia aos 30, 60, 90 e 120 dias após o transplantio. Os frutos foram colhidos regularmente para a avaliação do número total de frutos, número de frutos comerciáveis, número de frutos refugo, massa média dos frutos comerciáveis e refugo, diâmetro vertical e diâmetro horizontal, produção de frutos comerciáveis, produção de frutos refugo, e produtividade. A qualidade dos frutos foi mensurada pela análise de sólidos solúveis totais, pH da polpa, acidez titulável e relação entre sólidos solúveis e acidez titulável. A incidência da doença foi avaliada semanalmente após a averiguação da primeira planta murcha. Observações morfológicas e histológicas do calo cicatricial dos tratamentos avaliados foram realizadas. Todos os porta-enxertos avaliados apresentaram índice de pegamento ≥ 93,33%, indicando não haver incompatibilidade inicial entre as combinações avaliadas. O número de frutos comerciais diferiu significativamente nos porta-enxertos jiló comprido verde claro e cubiu, sendo este o tratamento com menor número total de frutos e número de frutos comerciais dentre os tratamentos avaliados. O cubiu apresentou os menores valores de produção de frutos comerciais e produtividade em relação aos tratamentos, com exceção da jurubeba juna em que não diferiu. Não houve incidência da murcha-bacteriana nos porta-enxertos jiló comprido verde claro e jurubeba juna, que conferiram resistência a doença nas plantas neles enxertadas. Os cortes histológicos evidenciaram características de incompatibilidade do tomateiro avaliado com os portaenxertos cubiu, jurubebão e jurubeba juna. Os porta-enxertos jiló comprido verde claro e jurubeba juna apresentaram alto índice de pegamento, resistência à murcha-bacteriana, e produtividade estatisticamente igual à testemunha nas condições do presente trabalho, de forma que possuem grande potencial para o uso como porta-enxertos na cultura do tomate. Palavras-chave: Solanaceae, Ralstonia solanacearum, Solanum lycopersicum, Resistência.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-08-22T19:15:30Z No. of bitstreams: 1 2013 - Evandro Silva Pereira Costa.pdf: 2748177 bytes, checksum: 3632d387d4cb90bea88d42726fae79ca (MD5) / Made available in DSpace on 2018-08-22T19:15:30Z (GMT). No. of bitstreams: 1 2013 - Evandro Silva Pereira Costa.pdf: 2748177 bytes, checksum: 3632d387d4cb90bea88d42726fae79ca (MD5) Previous issue date: 2013-12-18 / CAPES / The present work had the objective to characterize 59 accessions of tomato of the cherry group regarding its agronomic characteristics and resistance to the late blight and to select those more adapted to the organic agriculture. As standards were used: Carolina, Perinha ?gua Branca, Pending Yashi, Joanna and the hybrids Super Sweet, Sweet Million and Mascot F1. The experiments were conducted under field conditions in the Horticulture Sector of the Federal Rural University of Rio de Janeiro (UFRRJ) from June 2010 to November 2013 during which eight trials were carried out. In the different trials, productivity, number of total fruits, disease progression, morphological characteristics (coloration, shape, number of locules), physical (longitudinal and equatorial diameter), and physicochemical characteristics of fruits (total soluble solids - TSS , titratable total acidity - TTA, pH, TSS / TTA ratio). It was also evaluated the accumulation of dry mass of the stem, leaves and fruits, content and content of nitrogen (N), potassium (K) and phosphorus (P) in the respective organs. It was observed a great genetic variability between the accessions as to the physical and physical-chemical morphological attributes. Promising accesses were selected for use in breeding programs and with great productive potential and potential for cultivation in organic systems. Among these, we highlight the ENAS 1040, ENAS 1037, ENAS 1031 and ENAS 1026 accessions for cultivation in the spring / summer period and the accesses ENAS 1228, ENAS 1214, ENAS 1227 and ENAS 1220 by the highest total soluble solids (?Brix). The accessions ENAS 1121, EAS 1013, ENAS 1143 and ENAS 1029 were distinguished by the production of fruits with differentiated formats and the accesses ENAS 1007, ENAS 1008, ENAS 1037, ENAS 1033, ENAS 1017, ENAS 1036, ENAS 1062 and ENAS 1029 by coloring of the fruits. The accessions ENAS 1227 and 1026 stood out by the partial resistance to the late blight, equivalent to the standards 'Carolina' and 'Perinha ?gua Branca'. The accessions ENAS 1227, ENAS 1216, ENAS 1153 and ENAS 1060 were the ones that stood out the most regarding the resistance to the late blight and productivity. / O presente trabalho teve como objetivo caracterizar 59 acessos de tomateiro do grupo cereja quanto ?s suas caracter?sticas agron?micas e resist?ncia ? requeima e selecionar aqueles mais adaptados ? agricultura org?nica. Como padr?es foram utilizadas: Carolina, Perinha ?gua Branca, Pendente Yashi, Joanna e os h?bridos Super Sweet, Sweet Million e Mascot F1. Os experimentos foram conduzidos em condi??es de campo, no Setor de Horticultura da Universidade Federal Rural do Rio de Janeiro (UFRRJ) no per?odo de junho de 2010 a novembro de 2013 durante o qual realizaram-se oito ensaios. Nos diferentes ensaios, avaliaram-se produtividade, n?mero de frutos totais, progresso da requeima, caracter?sticas morfol?gicas (colora??o, formato, n?mero de l?culos), f?sicas (di?metro longitudinal e equatorial), e f?sico-qu?mica dos frutos (s?lidos sol?veis totais - SST, acidez total titulav?l - ATT, pH, rela??o SST/ATT). Avaliou-se, ainda, ac?mulo de massa seca do caule, folhas e frutos, teor e conte?do de nitrog?nio (N), pot?ssio (K) e f?sforo (P) nos respectivos ?rg?os. Observou-se grande variabilidade gen?tica entre os acessos quanto aos atributos morfol?gicos f?sicos e f?sico-qu?micos. Selecionaram-se acessos promissores para uso em programas de melhoramento e com grande potencial produtivo e com potencial para cultivo em sistemas org?nicos. Dentre estes, destacam-se os acessos ENAS 1040, ENAS 1037, ENAS 1031 e ENAS 1026 para cultivo no per?odo de primavera/ver?o e os acessos ENAS 1228, ENAS 1214, ENAS 1227 e ENAS 1220 pelos maiores teores de s?lidos sol?veis totais (?Brix). Os acessos ENAS 1121, EANAS 1013, ENAS 1143 e ENAS 1029 destacaram-se pela produ??o de frutos com formatos diferenciados e os acessos ENAS 1007, ENAS 1008, ENAS 1037, ENAS 1033, ENAS 1017, ENAS 1036, ENAS 1062 e ENAS 1029 pela colora??o dos frutos. Os acessos ENAS 1227 e 1026 destacaram-se pela resist?ncia parcial ? requeima, equivalente ? dos padr?es ?Carolina? e ?Perinha ?gua Branca?. Os acessos ENAS 1227, ENAS 1216, ENAS 1153 e ENAS 1060 foram os que mais se destacaram quanto a resist?ncia ? requeima e produtividade.

Se determinó la respuesta de resistencia de 10 patrones de tomate a una población avirulenta de Meloidogyne javanica en maceta. Los ensayos se realizaron en primavera, cuando las temperaturas permitían la expresión fenotípica de la resistencia del gen Mi-1 (28˚C), en verano sometidos a altas temperaturas y en campo, exponiéndolos a altas densidades poblacionales del nematodo. A temperaturas inferiores a 28˚C los patrones mostraron gran variabilidad en la respuesta de resistencia que osciló entre alta y moderadamente resistente (PG-76, Gladiator, MKT-410; Brigeor, 42851, 43965, Big Power y He-man), hasta susceptible (Beaufort, Maxifort). Por encima de 28˚C, sólo dos patrones (PG-76 y He-man) inhibieron la reproducción del nematodo. Frente a distintas poblaciones de M. arenaria, M. incognita y M. javanica, el patrón PG-76, fue altamente resistente a todas las poblaciones, Brigeor osciló entre altamente resistente y moderadamente resistente, mientras que Beaufort y Maxifort mostraron menor resistencia o fueron totalmente susceptibles; además ésta varió en función de la población analizada. Se caracterizó molecularmente el locus Mi-1 en los patrones híbridos y cultivares de tomate estudiados. Se emplearon los marcadores moleculares PM3, PMi y Mi23, específicos para la caracterización del locus Mi en patrones híbridos de tomate (S. lycopersicum × S. habrochaites; S. lycopersicum × S. chilense), mediante PCR. También se realizaron análisis bioinformáticos con marcadores específicos (Mint-up/do, C172, C2S4, IMO-F1/R1, y VIGS) para determinar la presencia del gen Mi-1.2 en dichos patrones. Los resultados mostraron que los marcadores PMi y Mi23 amplifican homólogos del gen Mi-1 en S. chilense, S. habrochaites y S. peruvianum y también en S. lycopersicum (marcador Mi23). El marcador PM3 amplificó el gen Mi-1.2 en Beaufort y Maxifort (S. lycopersicum × S. habrochaites) pero no fue efectivo para los híbridos de S. chilense. El marcador molecular PM3, no pudo determinar la expresión de del gen Mi-1.2 en Beaufort y Maxifort por hallarse fuera de la secuencia codificadora (CDS) del gen. Análisis bioinformáticos indicaron que ningún marcador específico diseñado para el gen Mi-1.2, podía este gen de otros homólogos presentes en S. lypcopersicum y S. peruvianum. El nuevo marcador Pau-Do en combinación con el primer C2S4, amplificaron un fragmento de 1.494 pb en la CDS del gen Mi-1.2 en raíces y hojas de Beaufort y Maxifort. La durabilidad de la resistencia del gen Mi-1 después del cultivo reiterado de patrones de tomate se determinó en ensayos de campo durante tres años consecutivos, empleando PG-76 y Brigeor. El patrón PG-76 fue muy resistente después del 1er ciclo de cultivo, pero mostró resistencia intermedia y suscetibilidad al finalizar el 2o y el 3er año de cultivo, respectivamente. El patrón Brigeor y el cultivar de tomate resistente Monika (control) mantuvieron un nivel de resistencia intermedio al final del 3er cultivo, aunque ensayos posteriores confirmaron la aparición de virulencia. Los resultados mostraron que el cultivo reiterado de patrones de tomate resistentes seleccionó rápidamente aislados virulentos de M. javanica. El fenotipo virulento de estas poblaciones se analizó molecularmente con el marcador MVC, diseñado para distinguir las poblaciones virulentas seleccionadas de Meloidogyne de los aislados naturalmente virulentos. Se analizaron dos poblaciones japonesas seleccionadas de M. incognita y M. javanica, tres poblaciones españolas virulentas seleccionadas, una población naturalmente virulenta y una avirulenta (todas M. javanica). Las muestras de ADN se obtuvieron de individuos juveniles o de hembras adultas y se incluyeron muestras de agua sin nematodos (5 µm filtrada) procedentes del drenaje de una maceta con una planta infectada por una población virulenta japonesa. El marcador MVC amplificó ADN en las muestras de agua pero no en las que sólo contenían ADN de nematodos. Las secuencias de ADN mostraron una estrecha correlación con diversas proteínas de especies de betaproteobacterias. Los experimentos revelaron que el marcador de MVC no está relacionado con un gen de virulencia del nematodo (avr) sino con betaproteobacterias. Finalmente, se estudió la existencia de homólogos del gen Mi en las especies de tomate silvestre Solanum chilense, S. habrochaites, S. peruvianum y S. huaylasense. La respuesta de resistencia de la variedad LA-1358 de S. huaylasense varió en función de la especie del nematodo estudiada: fue resistente frente a M. arenaria y susceptible frente a M. javanica. La reproducción de M. incognita fue muy variable y no difirió de la reproducción alcanzada en los dos cultivares empleados como controles. / The response of 10 Mi-1 tomato rootstocks to a Mi-avirulent population of M. javanica was determined in pot tests conducted in a greenhouse in spring when temperatures remained below the Mi-1 functionality resistance threshold (28 ˚C), and in summer when daily temperatures exceeded the Mi-1 expression threshold. Rootstocks were also evaluated in the field exposing them to high population densities of the nematode. Results on infectivity and reproduction below 28 ˚C indicated a wide variability in the resistance response of the rootstocks ranging from highly or intermediate resistance (PG-76, Gladiator, MKT-410; Brigeor, 42851, 43965, Big Power and He-man) to fully susceptible (Beaufort and Maxifort). At high temperature conditions, only PG-76 and He-man inhibited the reproduction of M. javanica. Rootstocks PG-76, Brigeor, Beaufort and Maxifort were challenged to different populations of M. arenaria, M. incognita and M. javanica. Rootstock PG-76 was highly resistant to all the populations tested, whereas the response of Brigeor ranged from highly to moderate resistance; the resistance response of rootstocks Beaufort and Maxifort varied according to the population tested. Molecular characterization of the resistance phenotype was performed for all the tomato hybrid rootstocks and cultivars tested. The markers PM3, PMi, Mi23, for the characterization of the Mi-locus of hybrid tomato rootstocks (S. lycopersicum × S. habrochaites and S. lycopersicum × S. chilense) were used for PCR reactions. In silico analyses were done with specific markers for the Mi-1.2 gene (Mint-up/do, C1/2, C2S4, IMO-F1/R1, and VIGS). Markers PMi and Mi23 were polymorphic for the Mi-1 locus in wild Solanum species (S. chilense, S. habrochaites, and S. peruvianum) and for S. lycopersicum (marker Mi23). Marker PM3 detected the Mi-1.2 gene in S. lycopersicum × S. habrochaites hybrid rootstocks, but not in the S. chilense hybrids. As marker PM3 is located outside the coding sequence of the Mi-1.2 gene, expression of this homolog could not be determined in Beaufort and Maxifort. In silico results indicated that none of the available markers for the Mi-1.2 gene could distinguish this homolog from the other Mi-homologs from S. lypcopersicum and S. peruvianum species. A new marker Pau-Do, in combination with C2S4, was designed to amplify in CDS of the Mi-1.2 gene. Amplification with these primers of cDNA from Beaufort and Maxifort indicated that the Mi-1.2 gene was expressed in both rootstocks, despite their susceptible phenotypic response to some Meloidogyne populations. The durability of the Mi-1 gene after repeated cultivation of resistant tomato rootstocks (PG-76 and Brigeor) was determined through field trials during three consecutive years. Rootstock PG-76 responded as highly resistant after the first cropping cycle, although it became fully susceptible after the second and the third cropping cycles. Rootstock Brigeor and the resistant tomato cultivar Monika (control), retained intermediate resistance levels at the end of the third year. Bioassays confirmed that selection of virulence occurred more rapidly in plots with rootstock PG-76 followed by Brigeor and the resistant tomato cultivar Monika. The virulent phenotype of the selected M. javanica populations in the field experiments was determined with MVC molecular marker, designed to distinguish selected from naturally virulent populations of Meloidogyne spp. The populations analyzed included two Japanese selected virulent populations, and the three virulent populations selected in the field trials, and one naturally virulent population and one avirulent population from Spanish. DNA samples were obtained from individual juveniles (J2) or adult females from all the selected virulent populations. Experiments included water samples free of nematodes (5-µm filtered), obtained from the draining-water of a plant infected by a Japanese selected virulent population. Amplification of DNA only occurred in samples of filtered water, but not in those containing only nematode genetic material. Sequencing and BLAST of the DNA fragments amplified by the MVC molecular marker, established a strong correlation of the amplified bands with proteins from betaproteobacteria species Overall, these results showed that the MVC marker is not related to a nematode virulence gene (avr) but to betaproteobacteria. New root-knot nematode resistant Mi-homologs were searched in accessions of the wild Solanum species. The S. huaylasense accession LA-1358 reduced reproduction of a population of M. arenaria to similar levels than the resistant tomato cultivar Anairis. Nevertheless, the resistance response of S. huaylasense accession LA-1358 was also nematode-species specific.

Solasodine, a steroidal alkaloid, is used by the pharmaceutical industry in certain parts of the world, as a raw material in the synthesis of steroid drugs. The compound is contained in many members of the genus Solanum, including S. mauritianum Scop., a common weed in South Africa. The levels of solasodine in three culture systems of S. mauritianum under various cultural conditions were examined. A high performance liquid chromatographic (HPLC) method was developed for the detection of solasodine. In order that low-cost, fixed wavelength ultra-violet detectors could be used, which would make the technique more widely applicable, a derivatization step, namely benzoylation, was included in the sample preparation. An extraction and purification protocol was then established, that complemented the HPLC technique and allowed successful detection of solasodine levels in a whole range of different sample types, including callus, suspension cultured cells, roots, stems and leaves. The three culture systems examined were callus, suspension and hairy root cultures. The callus system was used to establish which cultural parameters affected solasodine content in vitro to the greatest extent. A control culture was grown on a MURASHIGE and SKOOG (1962) medium (excluding glycine) supplemented with 3 % sucrose, 0.1 g I ¯¹ myo-inositol and lacking hormones. This culture contained an average of 9.2 μg g ¯¹ DW of solasodine. Many factors, including alteration of the carbon : nitrogen ratio and substitution of Gelrite for agar as the gelling agent, had no significant effect on the solasodine content of the callus or its growth. Greatly increased solasodine productivity of the callus was recorded when glucose was substituted for sucrose, the medium strength was reduced by half, or certain combinations of the hormones benzyladenine and naphthaleneacetic acid were added to the medium. The maximum levels of solasodine recorded in these cultures, on a per gram dry weight basis, equalled those of the vegetative parts of an intact S. mauritianum plant, but were approximately three times lower than those of the green berries. Suspension cultures could not be grown on the same medium as the callus cultures. Substitution of the vitamin complement of MURASHIGE and SKOOG (1962) with the so-called RT vitamin complement of KHANNA and STAB A (1968), resulted in successful growth and maintenance of S. mauritianum suspension cultures. The auxin, 2,4-dichlorophenoxyacetic acid (1 mg I ¯¹) was included in the medium. None of the suspension cultures grown on this medium, or slight modifications thereof, contained any trace of solasodine. This system could therefore not be used for the synthesis of solasodine in vitro. Hairy root cultures were initiated by inoculation of an excised hypocotyl of an in vitro-grown seedling of S. mauritianum with a 48 hour culture of Agrobacterium rhizogenes LBA 9402. Transformation frequency was extremely low. The transformed roots could be excised and grown successfully on a phytohormone-free medium, either in the solid or liquid form. Solasodine was extracted from hairy roots grown in a full-strength liquid MURASHIGE and SKOOG (1962) medium (excluding glycine) supplemented with 3 % sucrose and 0.1 g I ¯¹ myo-inositol, a half-strength such medium and a full-strength medium with 3 % glucose substituting for 3 % sucrose. In the latter medium, growth was very poor, whereas in the other two media, growth was very rapid . Both solasodine content (126 μg g¯¹DW) and root growth were greatest in the full-strength medium supplemented with 3 % sucrose. This level of solasodine was greater than that found in any of the callus cultures or vegetative parts of the plant and approached that of the green berries of S. mauritianum. Overall, of the culture types of S. mauritianum tested, the hairy root culture system appears to be most favourable for the in vitro production of solasodine. / Thesis (Ph.D)-University of Natal, Pietermaritzburg, 1993.

Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1992. / Vita. Abstract. Includes bibliographical references (leaves 53-54). Also available via the Internet.

Submitted by José Sidnaldo Pinsetta Junior (j.pinsetta@gmail.com) on 2018-09-10T13:07:04Z No. of bitstreams: 1 Dissertação José Sidnaldo final - repositorio.pdf: 4492049 bytes, checksum: 4ab3eb4e73554043616e04c8821af820 (MD5) / Approved for entry into archive by Neli Silvia Pereira null (nelisps@fcav.unesp.br) on 2018-09-10T18:26:39Z (GMT) No. of bitstreams: 1 pinsettajunior_js_me_jabo.pdf: 4492049 bytes, checksum: 4ab3eb4e73554043616e04c8821af820 (MD5) / Made available in DSpace on 2018-09-10T18:26:39Z (GMT). No. of bitstreams: 1 pinsettajunior_js_me_jabo.pdf: 4492049 bytes, checksum: 4ab3eb4e73554043616e04c8821af820 (MD5) Previous issue date: 2018-07-12 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A berinjela é uma olerícola de grande importância em diversos países e seu consumo tem aumentado no Brasil devido às características nutricionais para a alimentação humana. No entanto, esse vegetal apresenta limitações de comercialização como produto minimamente processado devido ao rápido escurecimento enzimático após o corte. O objetivo deste trabalho foi estudar os efeitos do recobrimento comestível a base de hidroxipropilmetilcelulose (HPMC) combinado, ou não, com agentes antiescurecimento sobre a qualidade de berinjelas minimamente processadas (BMP). Foram utilizadas berinjelas da cv Nápoli higienizadas e processadas em cubos, com posterior aplicação os recobrimentos por aspersão. Na primeira etapa foram testadas três formulações de HPMC + Cera de Abelha (CA) nas concentrações de 20, 40 e 60%. No segundo experimento testou-se o efeito do HPMC associado ao ácido cítrico (0,5; 1 e 1,5 %) e no terceiro experimento, ao ácido ascórbico (0,5; 1 e 1,5 %). As BPM foram acondicionadas em embalagens de PET (Polietileno tereftalato) e armazenadas em expositores refrigerados a 5°C. As análises foram realizadas a cada 3 dias até 12 dias, determinando-se a perda acumulada de massa fresca, firmeza, índice de brancura, composição gasosa do interior da embalagem, determinação de acetaldeído e etanol, compostos fenólicos totais, atividade das enzimas polifenoloxidase (PPO), peroxidase (POD) e fenilalanina amônia-liase (PAL), e contagem microbiana (microrganismos aeróbios mesófilos, coliformes totais e E. coli). O recobrimento com HPMC+40% de cera de abelha reduziu a atividade de enzimas responsáveis pelo escurecimento e a adição de 0,5% de ácido cítrico ou 1% de ácido ascórbico ao recobrimento levou a uma menor síntese de compostos fenólicos e menor atividade enzimática. / The eggplant is a vegetable of great importance in many countries and it has an increasing consumption in Brazil thanks to nutritional benefits for human health. Nevertheless, it presents limitations to commercialization due to fast enzymatic browning after cutting. The aim of this Project was to study the effects of an edible coating Hydroxypropyl methylcellulose (HPMC) based in association, or not, with food aditives on the quality of fresh-cut eggplant (FCE). Eggplants cv. "Napoli" were sanitised and processed in cubes of 2,5 x 2,5 x 2,5 cm and later sprayed with the coatings. In the first step, three HPMC and Beewax (BW), 20, 40 and 60% emulsions were tested. In the second experiment, the effect of HPMC+40% BW associated with citric acid (0,5; 1 e 1,5 %) were assessed and in a third experiment, in association with ascorbic acid (0,5; 1 e 1,5 %). The FCE was packed in PET trays and stored at 5°C. Analysis were carried out each 3 days until 12 days. It was assessed the accumulated loss of fresh matter, firmness, whiteness index, atmosphere inside packaging, acetaldehyde and ethanol determination, total phenols, enzyme activity of polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia-lyase (PAL) and microbiology counting (total mesophilic aerobic count, total coliforms and E. coli). The coating with HPMC+40% BW reduced the activity of enzymes responsible for browning and the addition of 0,5% of citric acid or 1% of ascorbic acid to the coating led to a lower synthesis of phenolic compounds and to lower enzymatic activity. / FAPESP: 2016/23600-1

Microtubers were obtained from potato plants (Solanum tuberosum L.) cultured in vitro, they were used in following experiments. The impact of growth regulators (FLD, AgNO3, BA, ABA) was monitored in length of dormancy. The content of ABA in the budding tubers and the content of endogenous CK (BA, IP, DHZ, DHZR, Z) was determined during the dormancy as well. Production of ACC, ethylene, O2, CO2 and ethane was determined by gas chromatography. Variants containing FLD, AgNO3 and BA had a significant impact in the shortening of dormancy and stimulation the growth of buds microtubers. When they were used the occurrence of budding tubers was increased by 30-40 % over the control. Variant of ABA inhibited the growth of buds. ABA content correlated with the process of dormancy and the occurrence of budding tubers. The highest content of ABA was in variant with freshly collected dormant tubers. Concentration of various CK was dependent on the type of CK and monitored variant. Generally, It was slightly increased with occurrence of budding tubers.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-08-31T16:43:51Z No. of bitstreams: 1 texto completo.pdf: 1571624 bytes, checksum: 532bd28b48be76bfcc011401faf8fee6 (MD5) / Made available in DSpace on 2017-08-31T16:43:51Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1571624 bytes, checksum: 532bd28b48be76bfcc011401faf8fee6 (MD5) Previous issue date: 2017-07-13 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O cultivo do tomateiro possui relevante importância sócio-econômica na agricultura brasileira. Mesmo diante deste cenário favorável, a tomaticultura é considerada uma atividade de elevado risco econômico e de grande complexidade agronômica. Dentre os principais problemas, destaca-se a requeima, causada pelo oomiceto Phytophthora infestans (Mont.) de Bary. O controle da doença é altamente dependente do uso de agrotóxicos, pois a maioria dos cultivares de tomateiro não são resistentes. Na tentativa de reverter esse quadro, a principal alternativa é a transferência de genes de resistência presentes em acessos de tomateiro conservados em Bancos de Germoplasma. Assim, após oito anos de pesquisas foi selecionado o acesso BGH-2127 (Solanum lycopersicum), como fontes de resistência a requeima, dentre os 870 acessos de tomateiro do Banco de Germoplama de Hortaliças da Universidade Federal de Viçosa (BGH-UFV). Pelo fato da resistência ser quantitativa, torna-se difícil a avaliação e seleção fenotípica de genótipos superiores, feita apenas no campo ou em associação com marcadores moleculares genéticos tipo QTL. Neste contexto, a metodologia da análise proteômica torna-se uma alternativa viável, pois pode fornecer importantes informações e ferramentas para maior entendimento das rotas metabólicas da interação planta-patógeno e assim auxiliar no desenvolvimento de futuras estratégias para o desenvolvimento de cultivares de tomateiro resistentes. Com o auxílio de ferramentas proteômica, os objetivos deste estudo foram: analisar possíveis diferenças entre os proteomas constitutivos de genótipos de tomateiro contrastantes quanto a resistência à requeima; e identificar proteínas frente a infecção do patógeno que possam explicar possíveis mecanismos moleculares de resistência do tomateiro a esta doença. Foram avaliados o acesso BGH-2127 e o cultivar Santa Clara, resistente e suscetível à requeima, respectivamente. Para avaliação do proteoma constitutivo, as proteínas foram extraídas de amostras das folhas dos genótipos sem que tivesse ocorrido a inoculação, apenas comparando a constituição proteica do BGH-2127 e do Santa Clara. Na avaliação à resposta ao patógeno, as plantas foram inoculadas com uma mistura de esporângios de P. infestans, na concentração de 1x10 3 esporângios mL -1 , coletados em regiões da Zona da Mata Mineira, e as proteínas foram extraídas de folhas de cada genótipo inoculado e não inoculado (controle) coletadas nos tempos 0, 2 e 48 horas. Na análise proteômica foi utilizada a técnica da eletroforese bidimensional (2-DE), com a primeira dimensão (focalização isoelétrica) realizada em tiras IPG de 24 cm, pH 3-10, e a segunda dimensão em SDS-PAGE, em gel 12,5%T. Foram obtidos três géis por tratamento, correspondendo a três repetições biológicas. Em seguida, os géis foram revelados por coloração com azul de coomassie coloidal. Posteriormente as imagens dos géis foram digitalizadas usando o equipamento Image Scanner III e analisadas no software ImageMaster 2D Platinum 7.5. Spots com variação de sobreposição acima de 1,5 e ANOVA p<0,05 foram considerados diferencialmente abundantes. Os spots das proteínas diferencialmente abundantes foram identificados em espectrômetro de massas do tipo MALDI-TOF/TOF Ultraflex III. As listas de massas obtidas foram confrontadas contra os bancos de dados de proteínas obtidos do UNIPROT, com auxílio do software MASCOT e os resultados obtidos, validados pelo aplicativo SCAFFOLD com pelo menos 90% de probabilidade. As proteínas identificadas foram categorizadas funcionalmente usando o software Mapman. Uma vez listadas as proteínas diferencialmente abundantes para os genótipos BGH-2127 e Santa Clara foi realizada a construção de uma rede de interação proteína-proteína utilizando o software STRING. Ensaios de PCR em Tempo Real (RT-PCR) foram realizados para confirmar os níveis de abundância de algumas proteínas identificadas. O RNA total foi extraído de amostras de folhas e o cDNA obtido foi quantificado pelo equipamento SpectraMax M5 microplate/cuvette reader. A amplificação dos fragmentos alvo foi realizada utilizando o equipamento StepOneTM Real-Time PCR System e para a quantificação da expressão gênica foi utilizado o software REST. Na avaliação do proteoma constitutivo, 19 proteínas foram identificadas, e estão relacionadas com metabolismo e energia, fotossíntese, estresse e defesa e à transcrição. Aproximadamente 90% destas proteínas foram mais abundantes no cultivar Santa Clara. As proteínas endoquitinase ácida de 26 kDa e ribonuclease T2 foram mais abundantes no BGH-2127. Atividade enzimática confirmou maior abundância de quitinase no acesso BGH-2127 em relação à Santa Clara. Os padrões de expressão avaliados por PCR em tempo real diferiram dos resultados da análise proteômica. Na avaliação da resposta ao patógeno, 56 proteínas diferencialmente abundantes foram identificadas, sendo 39 referentes ao genótipo resistente e 17 ao suscetível. Estas proteínas foram categorizadas em grupos de funções biológicas de energia e metabolismo, fotossíntese, estresse e defesa, transcrição, outras proteínas e não caracterizadas. Para o acesso BGH-2127, proteínas do estresse oxidativo (2-cis peroxirredoxina BAS1 e 2-cis peroxirredoxina) e relacionadas à patogênese da família PR-5 (taumatina) tiveram os níveis de abundancia relativa aumentados nos tempos 0 e 48 horas após a inoculação, respectivamente, e foram consideradas importantes para o mecanismo de defesa deste genótipo. Os padrões de expressão avaliados por PCR em tempo real diferiram dos resultados da análise proteômica. Redes de interações proteína-proteína forneceram importantes informações sobre atividades celulares envolvidas na resistência do BGH-2127 à requeima. Diferenças na abundância das proteínas endoquitinase ácida de 26 kDa e ribonuclease T2 entre os proteomas constitutivos dos genótipos avaliados podem contribuir para o mecanismo de resistência do BGH-2127 à requeima. Proteínas relacionadas ao estresse oxidativo (PR-9) e família taumatina (PR-5) possuem papel importante no mecanismo de defesa do acesso BGH- 2127 resistente a requeima do tomateiro. / Growing tomatoes has significant economic and social importance in brazilian agriculture. Nevertheless, the tomato production is considered an activity of high economic risk and major agronomic complexity. Among the main problems with regard to pests and diseases, there is the late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary. Disease control is highly dependent on the use of pesticides and there is no resistant cultivars. The main strategy to reverse that one situation is the introgression of resistance genes present in tomato accessions stored in germplasm banks. So, after eight years of research, BGH-2127 access (Solanum lycopersicum) was selected as a source of resistance to late blight, among the 870 tomato accessions of Vegetable Germplasm Bank of the Federal University of Viçosa (BGH-UFV). Due to the resistance to be quantitative it becomes difficult to practice selection of superior genotypes, made only in the field or associated with genetic molecular markers. In this context, the methodology of proteomics becomes viable alternative to provide valuable information and tools to better understanding the different ways of plant-pathogen relationship, thus this methodology can help future proposals genetic strategies to develop resistance cultivars of tomatoes. With the help of proteomic tools, the objectives of this study were: to analyze differences between the constitutive proteomes of tomato genotypes contrasting in resistance to late blight; and identify proteins against infection of the pathogen that may explain possible molecular mechanisms of resistance of tomato to this disease. The BGH-2127 and Santa Clara cultivar, resistant and susceptible to late blight respectively, were evaluated. For the evaluation of the constitutive proteome, proteins were extracted from leaf samples of the genotypes without inoculation, only comparing the protein constitution of BGH-2127 and Santa Clara. In evaluating the response to the pathogen, the plants were inoculated with a mixture of sporangia of P. infestans sporangia at a concentration of 1 x 10 3 sporangia mL −1 from different regions of Zona da Mata. Proteins were extracted from leaves of each genotype inoculated and non-inoculated (control) collected at time 0, 2 and 48 hours. In the proteomic analysis, the two-dimensional electrophoresis technique (2-DE) was used, with the first dimension (isoelectric focusing) performed using a 24-cm strip containing an immobilized pH gradient (IPG) ranging from 3 to 10 and the second dimension was performed on SDS-PAGE 12.5%T. Three gels were obtained by treatment, corresponding to three biological replicates. Then the gels were stained with colloidal coomassie blue. Posteriorly the gels were scanned with the aid of Image Scanner III and analyzed using ImageMaster 2D Platinum 7.0 software. The expression values were considered significant according to the analysis of variance (ANOVA) at p<0.05. Proteins in which the abundance ranged at least 1.5 times were considered to be differentially abundant. Protein identification was performed using the MALDI- TOF/TOF Ultraflex III type mass spectrometer. The standard list of proteins was obtained UNIPROT, through the use of the MASCOT application. The result obtained by MASCOT was validated by the SCAFFOLD application, with a minimum of 90% probability. Identified proteins were functionally categorized using Mapman program. Once the differentially abundant proteins for BGH-2127 and Santa Clara genotypes were listed, the construction of a protein-protein interaction network was carried out using the STRING software. Real-time PCR assays (RT-PCR) were performed to confirm the levels of abundance of some identified proteins. Total RNA was extracted from leaf samples and the cDNA obtained was quantified by the SpectraMax M5 microplate/cuvette reader. The PCR reaction was performed using StepOneTM Real- Time PCR System. Quantification of gene expression was performed using REST software. In evaluation of the constitutive proteome, nineteen proteins were identified, which were then related to metabolism and energy, photosynthesis, transcription, stress and defenses. Approximately 90% of these proteins were more abundant in Santa Clara, a susceptible cultivar. Acidic 26 kDa endochitinase and ribonuclease T2 proteins were more abundant in BGH-2127 access. The enzymatic activity confirmed a greater abundance of chitinase in the BGH-2127 access as compared to the cultivar Santa Clara. Gene expression analyses by real time PCR demonstrated that the mRNA levels were not correlated with the respective protein levels. Abundance of the acidic 26 kDa endochitinase and ribonuclease T2 proteins in the constitutive proteomes of BGH-2127 may be associated with the answer to the resistance of this access. In evaluating the response to the pathogen, fifty-six differentially abundant proteins were identified, with thirty-nine referring to the resistant genotype and seventeen to the susceptible genotype. These proteins were categorized into functional groups of energy and metabolism, photosynthesis, stress and defense, transcription, other proteins and not characterized. For access BGH-2127, oxidative stress proteins (2-cis peroxiedoxin BAS1 and 2-cis peroxiredoxin) and thaumatin-like protein had levels of relative abundance increased at times 0 and 48 hours after respectively, and were considered important for the defense mechanism of this genotype. The expression standards evaluated by RT-PCR differed from the results of the proteomic analysis. Protein-protein interaction networks provided important information on cellular activities involved in the resistance of BGH-2127 late blight. Abundance of the acidic 26 kDa endochitinase and ribonuclease T2 proteins in the constitutive proteomes of BGH-2127 may be associated with the answer to the resistance of this access. Proteins related to oxidative stress (PR-9) and thaumatin-like family (PR-5) play an important role in the BGH-2127 access defense mechanism resistant to tomato late blight.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2016-01-07T08:08:29Z No. of bitstreams: 1 texto completo.pdf: 385459 bytes, checksum: f1b68d6dade6f1f37952de2eeedf571a (MD5) / Made available in DSpace on 2016-01-07T08:08:29Z (GMT). No. of bitstreams: 1 texto completo.pdf: 385459 bytes, checksum: f1b68d6dade6f1f37952de2eeedf571a (MD5) Previous issue date: 2015-07-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O selênio (Se) é elemento traço essencial às funções do corpo humano, como controle do metabolismo de hormônios e de doenças cardiovasculares, prevenção da doença de Kashin-Beck, além de ter ação anticarcinogênica. Os teores de selênio (Se) nas plantas estão relacionados com a disponibilidade deste elemento no solo e a capacidade de absorção e acúmulo pelas plantas. Com intuito de produzir alimentos nutricionalmente mais completos e saudáveis para consumo humano e animal, uma estratégia utilizada é a biofortificação. Assim, objetivou-se avaliar doses e formas de aplicação de Se para fortificar tubérculos de batata. Os experimentos foram conduzidos em casa de vegetação, em vasos de 8 dm-3 de solo e selenato de sódio como fonte de Se. O delineamento foi o inteiramente aleatorizado com cinco repetições e os tratamentos consistiram de cinco doses via solo (0; 0,12; 0,25; 0,50 e 1,0 mg dm -3de Se) e cinco concentrações via foliar (0; 0,10; 0,21; 0,31 e 0,42 % de Se). Foram avaliados sintomas de fitotoxicidade, alterações fisiológicas ou morfológicas nas plantas, produtividade, variáveis fotossintéticas, atividade antioxidante total nos tubérculos e teores de Se no solo, parte aérea e nos tubérculos de batata. A aplicação de Se via solo não influenciou as variáveis fotossintéticas, o crescimento das plantas e a produção de tubérculos. Por outro lado, a aplicação foliar causou fitotoxidade nas folhas e redução na produção e na matéria seca de tubérculos. A atividade antioxidante total nos tubérculos não apresentou diferença entre os tratamentos. A aplicação de Se, independente da forma de aplicação e dose testada, não apresentou efeito significativo quanto aos teores de P, Mg, Zn, Cu, Fe e Ca nos tubérculos. Entretanto, observaram-se diferenças significativas quanto ao teor de Se nos tubérculos, em ambas as formas de aplicação e nas menores doses testadas. Os tubérculos de batata são capazes de acumular Se de maneira eficiente e a parte aérea das plantas transloca este elemento para os tubérculos, mesmo sob níveis severos de fitotoxidade. Palavras chave: Solanum tuberosum, translocação, nutrição humana. / Selenium (Se) is an essential trace element the human body functions such as hormone metabolism and control of cardiovascular diseases, prevention of Kashin-Beck disease, and have anticarcinogenic action.The levels of Se in the plant are related to the availability of this element in the soil and the ability to uptake and accumulation, of plants. Aiming to produce more nutritionally complete and healthy foods for human consumption and animal, a strategy used is biofortification. This experiment aimed to evaluate doses and Se application forms able to fortify potato tubers. The experiments were conducted in greenhouse, in pots of 8 dm-3 of soil and sodium selenate as a source of Se. The design was completely randomized with five replications and the treatments consisted of five doses in the soil (0; 0.12; 0.25; 0.50 and 1.0 mg dm-3 Se) and five foliar concentrations (0; 0.10; 0.21; 0.31 and 0.42 % of Se). Were evaluated phytotoxicity symptoms, physiological or morphological changes in plants, productivity, photosynthetic variables, total antioxidant activity in the tubers and Se levels in the soil, shoot and potato tubers. The use of Se in the soil did not affect the photosynthetic variables, the growth of plants and tuber yield. Moreover, foliar application caused phytotoxicity on leaves and reduction in dry matter production and tubers. The total antioxidant activity in the tubers showed no difference between treatments. Applying Se, regardless of the form of application and dose tested, no significant effect as the P, Mg, Zn, Cu, Fe and Ca contents in tubers was observed. However, there were significant differences in the Se content of tubers in both application forms and in smaller doses tested. The potato tubers are able to accumulate Se efficiently and the shoot translocates this element to the tubers, even with severe levels of phytotoxicity. Keywords: Solanum tuberosum, translocation, human nutrition.