Thesis (MscAgric (Horticulture))--University of Stellenbosch, 2008. / Commercial production of sweet cherries has recently increased in South Africa, with more than 400 ha planted by 2006. Cherry, a high chilling fruit variety, is however not suited for the mild winter climate of South Africa. This was recognizable through common observed symptoms of delayed foliation and poor fruit set. In addition, cherry is exposed to long and hot summers in the postharvest period. The objective of this study was to evaluate cherry cultural practices that can manipulate (reduce) the trees chilling requirement under South African conditions. Cultural practices where aimed at increasing reserves (nitrogen, cytokinin and carbohydrates) in the tree. In addition, bud dormancy progression of cherry buds was quantified to determine the bud dormancy progression pattern under mild winter conditions. This was achieved through sampling of cherry shoots from different cherry production areas which was then forced in the growth cabinets. A model was developed to identify possible factors and groupings that can explain the cherry bud dormancy pattern. A model, comprising two joined straight lines, was fitted in order to characterize bud dormancy behaviour for sweet cherry cultivars under mild winter conditions. All cherry cultivars followed the expected pattern of entrance and exit from dormancy. Factor analysis showed that factors related to the entrance into dormancy primarily characterize bud dormancy behaviour. Bud dormancy patterns were also a function of environmental conditions within a year as shown by cluster analysis. In addition, buds entered dormancy in mid-summer and remained dormant until chilling accumulation commenced. Bud dormancy release was generally extended over a three to five-month period for all cultivars. Prior to spring budburst exit of both lateral and terminal buds occurred rapidly. Data indicate that there is no ecodormant phase for cherry under the prevalent climatic conditions in South Africa. Further experimentation was aimed at increasing reserves within the trees through cultural practices. In the nitrogen trials, fertilization in the postharvest period had no significant effect on field budburst or bud dormancy progression in one-year-old shoots. Time of flowering was advanced in N treatments during 2007 only. Yield was not significantly increased. Therefore, in this trial, N fertilization in the postharvest period did not significantly reduce the chilling requirement of mature sweet cherry trees under mild winter conditions. Application of particle films (Surround® and Raynox®) or ethylene inhibitors (Retain®) in the summer did not reduce the heat stress the trees experienced. Treatments had no significant effect on carbon assimilation, stomatal conductance, leaf surface temperature, fluorescence, bud dormancy, budburst, flowering and fruit set. Cytokinins sprays (benzyladenine) in autumn did not affect bud dormancy progression, spring budburst or flowering. Hydrogen cyanamide application in spring significantly advanced budburst, time to full bloom and increased yield. Promalin® and Retain®, however, had no significant effect on budburst, flowering or yield. It is therefore evident that cherry, due to its unexpected bud dormancy behaviour and its inability to be significantly influenced by several cultural practices, adapts poorly to South African climatic conditions through not reducing its chilling requirement significantly.

Thesis (MscAgric (Horticulture))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Commercial fig production with popular European cultivars, Bourjasotte Noire, Col de Damme Noire and Noire de Caromb, is new to the Western Cape. Little research on fig production has been conducted in South Africa and producers are struggling to implement effective commercial practices. In order to establish practices that will maximise yield of quality fruit, the most productive one-year-old shoot lengths were identified in a phenological study. All shoot length categories evaluated in ‘Bourjasotte Noire’ (10 – 15 cm, 25 – 40 cm, 50 – 65 cm, 75+ cm) yielded fruit and will probably yield well the following season. In ‘Col de Damme Noire’, shoots longer than 60 cm seem to be suited to reproduction, yet they might produce a poor yield the following season. Shoots 10 – 20 cm long in ‘Noire de Caromb’ are productive relative to their length, while shoots 30 – 50 cm and 60 – 80 cm long are also fairly productive. Shoots longer than 100 cm produced suitable shoot lengths for yield the following season. Two experiments were conducted on ‘Bourjasotte Noire’ and ‘Col de Damme Noire’ to establish the type (Experiment 1, different intensity heading cuts) and timing (Experiment 2) of pruning cuts required to stimulate the growth of shoots of the same length as the shoots selected to be optimal for yield in the phenology study, and to reduce the expression of distal branching. In ‘Bourjasotte Noire’, removing one third of the total length of one-year-old shoots on 21 July by heading stimulated the development of more growth and longer current season shoots compared to other treatments, while reducing yield slightly. Heading back to three nodes in ‘Col de Damme Noire’ stimulated the growth of current season shoots of the optimal length established in the phenology study, while heading cuts on 30 June produced the longest average current season shoot length in ‘Col de Damme Noire’. To further address the effects of distal branching (acrotony), an experiment was conducted to establish whether rest breaking agents (RBA’s) in combination with tip-pruning can increase tree complexity by improving bud break, and whether increased complexity would increase yield in all three cultivars. A second experiment was conducted to evaluate the effects of timing RBA applications on bud break and harvest scheduling. Lift® increased the number of buds breaking in ‘Bourjasotte Noire’, while in ‘Noire de Caromb’ Dormex® and oil increased bud break. Tip-pruning increased the average shoot length in ‘Bourjasotte Noire’ and ‘Col de Damme Noire’, while causing a reduction in the amount of new growth in ‘Noire de Caromb’. Lift® applied 3 August and Dormex® applied 30 June shortened the number of days to 50% bud break in ‘Bourjasotte Noire’ and ‘Col de Damme Noire’. Both Lift® and Dormex® applied on 30 June decreased the number of days to 50% bud break and 50% harvest of the breba crop in ‘Noire de Caromb’. These treatments increased the number of fruit in both the breba and main crop of ‘Noire de Caromb’, but reduced fruit size. In conclusion, different approaches with regards to pruning needs to be followed for each cultivar to establish or maintain the optimal shoots for reproduction, while RBA’s can be used to force earlier, increased- bud break and harvest of breba fruit. / AFRIKAANSE OPSOMMING: Kommersiele verbouing van drie Europese vykultivars, Bourjasotte Noire, Col de Damme Noire en Noire de Caromb is nuut tot die Wes-Kaap. Baie min navorsing oor die verbouing van vye is al in Suid-Afrika gedoen, met die gevolg dat produsente sukkel om effektiewe kommersiële verbouingspraktyke te implementeer. ‘n Fenologiese studie van die drie kultivars is uitgevoer om vas te stel wat die mees produktiewe een-jaar-oue lootlengte is, met die doel om die opbrengs van kwaliteit vrugte te maksimeer. Al vier kategorieë wat ge-evalueer is in ‘Bourjasotte Noire’ (10 – 15 cm, 25 – 40 cm, 50 – 65 cm, 75+ cm) is gevind om geskik te wees vir huidige en volgende seisoen opbrengs. Lote langer as 60 cm is geskik vir opbrengs in ‘Col de Damme Noire’ in die huidige seisoen, maar mag in die volgende seisoen swak presteer a.g.v. die gebrekkige lengte van nuwe lote wat daarop ontwikkel. In ‘Noire de Caromb’ is gevind dat lote 10 – 20 cm lank baie produktief is relatief tot hul lengte en dat lote 20 – 50 cm en 60 – 80 cm lank ook relatief produktief is. Lote langer as 100 cm was minder produktief, maar het nuwe lote gelewer wat geskik is vir opbrengs die volgende seisoen. Twee snoei eskperimente is uitgevoer op ‘Bourjasotte Noire’ en ‘Col de Damme Noire’ om vas te stel wat die mees geskikte tipe snoeisnit (Eksperiment 1, verskillende dieptes van topsnitte) en tydstip om te snoei (Eksperiment 2) is met die doel om lote te produseer soortgelyk in lengte aan die wat in die fenologie studie uitgewys is as die produktiefste, en om moontlik die voorkoms van “kaalnekke” te verminder. In ‘Bourjasotte Noire’ is gevind dat die wegsnoei van ‘n derde van die loot op 21 Julie aanleiding gee tot meer groei, langer een-jaar-oue lote en ‘n effense afname in opbrengs. Geskikte lote langer as 60 cm kan in ‘Col de Damme Noire’ verkry word deur lote te top sodat net drie nodes oorbly. Die uitvoer van topsnitte op 30 Junie het langer gemiddelde lootlengtes tot gevolg gehad. Om die probleem van “kaalnekke” (apikale dominansie) verder aan te spreek, is ‘n eksperiment uitgevoer om vas te stel of rusbreekmiddels gekombineerd met tip-snoei gebruik kan word om kompleksiteit te vermeerder deur knopbreek te verhoog, en indien wel, of dit sal aanleiding gee tot verhoogde opbrengs in al drie kultivars. ‘n Tweede eksperiment met verskillende toedieningstye van rusbreekmiddels is uitgevoer om vas te stel of oeste geskeduleer kan word. Lift® het knopbreek verhoog in ‘Bourjasotte Noire’, terwyl Dormex® en olie knopbreek verhoog het in ‘Noire de Caromb’. Tip-snoei het die gemiddelde lootlengtes verhoog in ‘Bourjasotte Noire’ en ‘Col de Damme Noire’, terwyl dit groei verminder het in ‘Noire de Caromb’. Lift® toediening op 3 Augustus en Dormex® toediening op 30 Junie het die aanvang van 50% knopbreek vervroeg in ‘Boujasotte Noire’ en ‘Col de Damme Noire’. Beide Lift® en Dormex® toediening op 30 Junie het die bereiking van 50% knopbreek- en 50% oes vervroeg in ‘Noire de Caromb’. Hierdie behandeling het ook die aantal vrugte van die breba- en hoofoes vermeerder, maar vruggrootte verminder. Verskillende snoeibenaderings behoort dus gevolg te word om vir elke kultivar die regte lootlengtes te genereer of te onderhou, terwyl rusbreekmiddels gebruik kan word om knopbreek te vervroeg en verhoog, en om die breba oes te vervroeg.

Made available in DSpace on 2014-08-20T13:25:41Z (GMT). No. of bitstreams: 1 TESE_Valtair_Verissimo.pdf: 3590695 bytes, checksum: 0329fe51ba9b30e95021c9dd6b0ef7b5 (MD5) Previous issue date: 2008-08-18 / Floral bud abortion has been the main limiting factor to the expansion of the pear crop in Brazil. That floral bud abortion has been considered as a complex problem that may be affected by physiological, sanitary, genetic and climatic factors. The rootstock that is the main subject of this study could also be an influencing factor in pear floral abortion, since until the year 2000, only the vigorous rootstocks Pyrus calleryana and Pyrus betulaefolia were used in Brazil. After that year, the pear growers of Santa Catarina and Rio Grande do Sul states started using dwarfing quince rootstocks such as Adams and EMC and the results has been satisfactory. In general these quince rootstocks are more sensitive to water stress during drought periods, since they develop a relatively small root system. However, the influence of these rootstocks on the physiology of dormancy, carbohydrate metabolism and floral biology are not known. With the objective of studying such unknown aspects, five experiments were carried out from 2005 to 2007 growth seasons. Part of these experiments was conduced under field conditions in a commercial orchard in Vacaria, RS, whereas other experiment activities were developed at Embrapa Clima Temperado research center in Pelotas, RS. All were composed of: a general literature review on the pear bud abortion subject, and plus five scientific articles, each one corresponding to one of the five above referred experiments. The first article contain the biological method to determine the dormancy level, on the pear cultivars Abbe Fetel and Packham s Triumph grafted on the quince rootstock EMC or Adams. The second article brings the results on carbohydrate levels in tissues of pear trees. The third article brings the results on floral biology of the above referred two cultivars on the two above referred quince rootstocks. The fourth article reports the results on the chilling requirement of potted trees of the pear rootstocks quinces EMC, EMA and Adams and of the pear rootstocks Pyrus calleryana and Pyrus betulaefolia, all them grown under controlled conditions. The fifth article reports the effects of chilling on potted trees of the pear cultivars Kieffer and Packham s Triumph grafted on the rootstocks quinces Adams, EMC or EMA, or on Pyrus betulaefolia, under controlled conditions. It were concluded that: the sugar transport and the floral biology were affected by rootstocks but not the dormancy development; the plants of the pear rootstocks quinces EMA, EMC and Adams have higher chilling requirements than the Pyrus calleryana and Pyrus betulaefolia rootstocks; the rootstock used changes the vegetative budbreak percentage of canopy cultivar; and that the higher chilling hours accumulated increased budbreak percentage and reduced floral bud abortion only until the chilling requirement is attained but floral bud abortion is increased after that condition. / O abortamento floral em pereira é o principal problema que limita o desenvolvimento da pereira no Brasil, caracterizado por um complexo de fatores fisiológicos, fitossanitários, genético e ambiental. O porta-enxerto, objeto central deste trabalho, pode ser um fator que tem contribuído para aumentar tal problema, pois a cultura da pereira se baseou, ao longo das últimas três décadas, no uso dos porta-enxertos Pyrus calleryana e P. betulaefolia. A partir de 2000 os produtores do RS e SC passaram a utilizar os marmeleiros, principalmente o Adams e EMC , obtendo resultados satisfatórios. Em geral esses porta-enxertos são mais sensíveis ao estresse hídrico, por apresentarem sistema radicular pouco desenvolvido. Entretanto se desconhece a influência dos mesmos na fisiologia da dormência, na mobilização de carboidratos durante o inverno e na biologia floral. Para atingir tais objetivos foram conduzidos cinco experimentos entre 2005 e 2007, sendo parte deles conduzidos em pomar comercial, na empresa Frutirol, em Vacaria-RS, e em condições controladas, na Embrapa Clima Temperados, em Pelotas-RS. A tese possui seis capítulos, sendo o primeiro uma caracterização e revisão bibliográfica geral do assunto e os demais cinco artigos científicos. O primeiro artigo teve como objetivo avaliar a dinâmica da dormência, através do uso do método biológico, de duas cultivares de pereira a Abate Fetel e a Packham´s Triumph enxertadas em marmeleiros EMC e Adams . O segundo visou determinar a concentração de carboidratos solúveis e amido. O terceiro teve por objetivo o estudo da biologia floral. O quarto artigo, conduzido em condições controladas, com plantas em vasos, tratou da determinação da exigência em frio dos porta-enxertos de marmeleiros EMC , EMA e Adams , além dos porta-enxertos do gênero Pyrus (Pyrus calleryana e P. betulaefolia). O quinto artigo, também em condições controladas, visou determinar o efeito do frio em plantas conduzidas em vaso nas cvs. Kieffer e Packham´s Triumph, enxertadas em Adams , EMC , EMA e Pyrus betulaefolia . Conclui-se que a mobilização de açúcares e amido e a biologia floral são influenciadas pelo tipo de porta-enxerto, mas não alterou a dinâmica da dormência. Nos experimentos conduzidos com plantas em vaso, foi possível identificar que os marmeleiros Adams , EMA e EMC possuem requerimento em frio maior do que P.calleryana e P. betulaefolia. O porta-enxerto utilizado altera a percentagem de brotação da cultivar copa, devido a redução do requerimento em frio ou calor. Por fim, identificou-se que o maior acúmulo de frio aumentou a brotação e reduziu a incidência de abortamento de gemas florais, mas somente até suprir o requerimento das plantas; quando em excesso foi prejudicial.

A perda do parênquima hepático, induzida por tratamento agudo, cirúrgico ou químico, desencadeia um processo regenerativo até que a massa hepática seja completamente restaurada. A regeneração hepática, após a hepatectomia parcial de 70%, é um dos modelos mais adequados de regeneração de células, órgãos e tecidos mais estudados. No fígado, ainda que sejam atribuídas propriedades regenerativas, muitas das lesões são tão prejudiciais que este mecanismo de reparação é insuficiente, tornando o transplante a única opção. As células derivadas do broto hepático de ratos apresentam uma boa alternativa para tratamento de doenças hepáticas devido ao seu alto índice proliferativo e da expressão de marcadores de pluripotência, sendo sua aplicabilidade viável em modelos experimentais. O objetivo deste trabalho foi analisar as diferentes vias de administração das células derivadas do broto hepático de ratos com 12,5 dias de gestação visando a melhor regeneração do órgão. Foram realizados experimentos de hepatectomia parcial de 70% (N=50 animais), PET Scan MSFX PRO In-Vivo RX e fluorescência, índice hepatossomático, análise de marcadores solúveis (GH, AFP, CEA, IGF-1), análises hematológicas, microscopia de luz (coloração HE, Tricômio de Masson), análise de marcadores por citometria de fluxo (CD90, STRO-1, Nanog, Oct3/4, Ki-67, Caspase 3) e ciclo celular por citometria de fluxo. Nossos dados demonstraram que as células do broto hepático administradas na via endotraqueal apresentaram melhor equilíbrio entre proliferação e morte celular, com maior expressão dos marcadores de pluripotência, melhor organização celular e regeneração tecidual, em contraste com outras vias, incluindo endovenosa, intraperitoneal e oroenteral. Isto a torna mais segura e de maior viabilidade na regeneração celular em relação às demais vias, sendo mais eficiente nos modelos de hepatectomia parcial / The restoration of liver parenchyma after partial hepatectomy or chemical treatments represents appropriate models to study regeneration mechanisms. The most appropriate model for liver regeneration is partial hepatectomy of 70%, however, organ repair properties are insufficient, suggesting the transplantation the best alternative to treat liver diseases. Cells derived from liver buds of rats show a high proliferative index and the expression of pluripotency markers; thus their significance for regeneration purposes can be tested experimentally. We here investigated different routes to administer cells derived from rat live buds of 12.5 days of gestation to adult individuals (N=50 animals) suffering from partial hepatectomy (70%). Applied methods included PET Scan MSFX PRO In-Vivo RX, fluorescence hepatossomatic index, analysis of soluble markers (GH, AFP, CEA, IGF-1), hematological analysis, light microscopy (staining HE, and Masson trichrome) as well as flow cytometry for cell cycle analysis and CD90, STRO-1, Nanog, OCT3/4, Ki-67, Caspase 3 expression. Our data showed that administration via the tracheal route resulted as favorite in regard to the balance between proliferation and cell deaths, of pluripotency marker expression, cellular organization and tissue regeneration, in contrast to other routes including: intravenous, intraperitoneal and oroenteral. Consequently, the tracheal route showed safer and more efficient treatment to enhance cell regeneration after partial hepatectomy

A transformação genética possibilita a introdução de genes de interesse agronômico no genoma das plantas e pode ser empregada na tentativa de obter plantas resistentes a doenças. No entanto, para se obter uma planta transgênica é necessário primeiramente estabelecer um procotolo eficiente de regeneração de plantas in vitro. Assim, o objetivo desse trabalho foi estudar a organogênese in vitro e a transformação genética de limão Volkameriano e laranja azeda com um fragmento do gene da capa protéica do CTV. Para a organogênese in vitro utilizou-se, como explante, segmento internodal, obtido de planta cultivada em casa-de-vegetação, segmento de epicótilo, coletado de plântula cultivada in vitro e segmento de cotilédone associado ao hipocótilo obtido de semente introduzida in vitro. Esses explantes foram mantidos em meio de cultura EME suplementado com 6-benzilaminopurina (BAP 0,0; 0,5; 1,0; 1,5; 2,0 mg L- 1), sendo incubados sob fotoperíodo de 16 h de luz ou em condições de escuro por 30 dias e então transferidos para fotoperíodo de 16 h de luz. A avaliação foi realizada após 45 dias de cultivo, determinando-se o número de explantes responsivos e o número de gemas por explante. A caracterização anatômica do processo de regeneração foi realizada por meio de cortes histológicos. Pela análise dos dados foi possível verificar que a organogênese in vitro ocorreu a partir dos três tipos de explantes testados, sendo que, nas duas espécies em estudo, os melhores resultados foram obtidos com o cultivo de segmento de cotilédone associado ao hipocótilo. As concentrações de BAP que estimularam as melhores taxas de regeneração foram de 1,0 e 1,5 mg L-1, para limão Volkameriano, e 0,5 e 1,0 mg L-1 para laranja azeda. A incubação dos explantes em ausência de luz favoreceu a regeneração in vitro. Pela análise histológica foi possível observar que o processo de regeneração, a partir dos três tipos de explantes testados, ocorreu por meio de organogênese indireta. O protocolo de desenvolvimento estabelecido durante os experimentos de organogênese in vitro foi utilizado para a transformação genética dessas espécies via Agrobacterium, contendo o plasmídeo pCAMBIA 2201, com um fragmento do gene da capa protéida do CTV, em uma construção gênica do tipo hairpin. As gemas de limão Volkameriano e laranja azeda identificadas como transgênicas pelo teste histoquímico GUS foram enxertadas in vitro em citrange Carrizo. A confirmação da transformação genética foi realizada pela análise de PCR, a qual mostrou a amplificação de um fragmento de 671 pb, correspondente a parte do gene amplificada. / Genetic transformation permits the introduction of agronomically important genes in plant genome and can be utilized in order to produce disease resistant plants. However for the recovery of transgenic plants is required to establish an efficient in vitro plant regeneration protocol. In this work the aim was to study an in vitro organogenesis and the genetic transformation of Volkamer lemon and sour orange with a sequence of the CTV coat protein gene. For in vitro organogenesis explant internodal segments collected from plants cultivated in greenhouse, epicotyl segments obtained from in vitro cultivated seedlings and cotyledon fragment with hypocotyl attached obtained from in vitro germinated seed were used as explant. These explants were cultured in EME medium supplemented with benzilaminopurine (BAP 0,0; 0,5; 1,0; 1,5; 2,0 mg L-1). Cultures were maintained under a 16 h photoperiod or in the dark for 10 d and then transferred to a 16 h photoperiod. The evaluation was performed 45 d after the incubation determining the number of responsive explant and the number of buds per explant. The anatomical characterization of in vitro regeneration process was carried out through histological analyses. The in vitro organogenesis occurred in the three types of explant tested, however cotyledon fragment with hypocotyl attached showed higher morphogenetic potential in both species. The best responses of regeneration were obtained when the medium was supplementation with 1,0 e 1,5 mg L-1 BAP for the Volkamer lemon and 0,5 e 1,0 mg L-1 BAP for the sour orange. The incubation in darkness favored the in vitro regeneration. The histological analyses showed that the regeneration process occurred through indirect organogenesis in the three types of explants tested. The developed protocol was use for genetic transformation of Volkamer lemon and sour orange with Agrobacterium, containing pCAMBIA 2201 plasmid with a sequence of CTV coat protein gene, in a hairpin construction. Volkamer lemon and sour orange shoots identified as transgenic by histochemical test GUS were micrografted into Carrizo citrange. PCR analysis were performed after micrografted showing the presence of the 671 pb fragment of the transgene.

As células-tronco são capazes de se diferenciarem em praticamente todos os tipos celulares, podendo ser utilizadas em terapias de substituição em várias doenças. Células de linhagens hepáticas embrionárias e fetais pode ser uma fonte importante para a terapia celular em indivíduos com doenças hepáticas, pois possuem um alto índice de diferenciação em hepatócitos e células do ducto biliar. O objetivo deste trabalho foi avaliar o potencial e o controle da resposta proliferativa de células do broto hepático de ratos Fischer 344 em cultura. A obtenção das células do broto hepático foi realizada nos períodos 12,5; 14,5 e 16,5 dias de gestação e os fragmentos foram cultivados em meio DMEM-F12. A caracterização morfológica foi realizada em microscopia invertida de luz e eletrônica de varredura. A caracterização dos marcadores de células mesenquimais, do citoesqueleto, progressão e fases do ciclo celular, morte celular, e do potencial elétrico mitocondrial foram realizadas por citometria de fluxo. A função bioquímica foi realizada pela análise das transaminases hepáticas e pela produção de radicais livres lipoperoxidados. Os resultados encontrados no acasalamento dos ratos isogênicos da linhagem Fischer 344 seguiu o protocolo de 12 h de luz, alcançando o índice reprodutivo satisfatório e reprodutível. O isolamento e separação do broto hepático desta linhagem de ratos permitiu a obtenção, expansão e caracterização de uma cultura primária nos diferentes dias de gestação 12,5; 14,5 e 16,5. Os aspectos morfológicos encontrados foram de células fusiformes do tipo fibroblast-like para todos os períodos de gestação em cultura. As análises das transaminases hepáticas TGO e TGP, mostraram se em concentrações menores no período 12,5 dias de gestação. A expressão dos marcadores de células-tronco de origem mesenquimais (CD90, Nanog, Oct ¾ e STRO1), marcadores do citoesqueleto (CK8, CK18 e Desmina), marcadores envolvidos na checagem e progressão do ciclo celular (P53, P21, P27 e Ciclina D1) e marcadores envolvidos na morte celular (Anexina V/PI, Caspase 3, Bax, Bad e Bcl2) mostraram diferencialmente expressos, nos diferentes períodos gestacionais. A análise do potencial elétrico mitocondrial nos períodos de gestação de 14,5 e 16,5 dias, foi maior na proporção de células inativas com menor capacidade funcional ou metabólica, quando comparada ao período de 12,5 dias ou mesmo em relação ao hepatócito normal de um rato adulto. Análise das fases do ciclo celular observou-se uma concentração de célula na fase G0/G1, repercutindo na modulação da capacidade de proliferação e aumento na proporção de células com DNA fragmentado, nas CBH no período de 16,5 dias em comparação aos demais períodos e ao hepatócito normal. O aumento da fragmentação do DNA em média de 3,5 e 2,3 vezes, respectivamente nos períodos de 12,5 e 14,5 dias de gestação. A capacidade de síntese e de divisão celular das CBH foi mantida em todos os períodos de gestação. Os radicais livres lipoperoxidados mostraram quantidades insignificantes em CBH nos diferentes dias de gestação. / Stem cells are able to differentiate into virtually all cell types and can be used in replacement therapies in various diseases. Cells of embryonic and fetal liver lines can be an important source for cell therapy in patients with liver disease because they have a high rate of differentiation into hepatocytes and biliary duct cells. The aim of this study was to evaluate the potential and control of the proliferative response of liver bud cells during the maturation of hepatocytes in culture. The acquirement of liver bud cells was performed for 12.5, 14.5 and 16.5 days of gestation and the fragments were cultured in DMEM-F12. Morphological characterization was performed on inverted light microscopy and scanning electron microscopy. The characterization of mesenchymal cell markers, cytoskeleton, and progression stages of the cell cycle, cell death and mitochondrial electric potential were performed by flow cytometry. The biochemical function was performed by analysis of liver transaminases and the production of free radicals lipoperoxidados. The results found in rats\' mating isogenic Fischer 344 followed the protocol of 12 hours of light reaching the reproductive rate satisfactory. The isolation and separation of the liver bud of this strain of mice allowed the isolation, expansion and characterization of a primary culture at different days of gestation, 12.5, 14.5 and 16.5 days. The morphological features were found of spindle cell type to fibroblast-like cells of the liver bud in culture. Analyses of hepatic transaminases GOT and GPT showed lower concentrations in the period 12.5 days of gestation. The expression of stem cell markers of mesenchymal origin (CD90, Nanog, Oct STRO1 and ¾), cytoskeletal markers (CK8, CK18 and desmin) markers involved in checking and cell cycle progression (P53, P21, P27 and Cyclin D1) and markers involved in apoptosis (Annexin V / PI, caspase 3, Bax, Bad and Bcl2) showed differentially expressed in different gestational periods. In the analysis of mitochondrial electrical potential in the gestation periods of 14.5 and 16.5 days, it was observed the higher proportion of quiescent cells with lower metabolic of functional capacity when compared to the periodo of 12.5 days or even compared to a normal hepatocyte adult rat. In the analysis of cell cycle phases it was observed the concentration of cell in the G0/G1 phase, resulting in modulation of proliferation capacity and the increase in the proportion of cells with fragmented DNA, CBH in the period 16.5 days compared to other periods and the normal hepatocyte. The increase of DNA fragmentation on average 3.5 and 2.3 times respectively in the periods of 12.5 and 14.5 days of gestation. The synthesis and cellular division of CBH was maintained in all stages of pregnancy. The free radicals lipoperoxidados showed insignificant amounts of CBH in the days of gestation.

Dans un sens large, le statut azoté de l’arbre impacte largement sa physiologie au printemps. Cependant la plupart des recherches conduites sur la remobilisation du carbone et de l’azote en cette période ont négligé l’hypothèse d’un rôle significatif de l’absorption d’azote avant le débourrement, en particulier, celui agissant en interaction avec l’utilisation des réserves carbonées et azotées et qui interviendrait dans la qualité de la croissance des jeunes pousses. Cette recherche a été entreprise avec une approche expérimentale et a été conduite avec de jeunes peupliers hybrides (Populus tremula × Populus alba, clone INRA 717-1B4). Ils ont été utilisés en nombre limité, mais présentaient, par construction initiale, des architectures différentes. Trois séries d’expériences ont ainsi été conduites en trois ans : tout d’abord avec le scion d’un an, puis une petite souche de deux ans et enfin, en associant les deux types de structure, une petite souche portant des réitérations équivalentes au scion d’un an. Les essais ont été conduits en environnement contrôlé et ont fait appel à trois régimes de fertilisation azotée appliquée en solution nutritive recyclée et pendant la transition « repos végétatif et reprise de croissance foliaire » : une fertilisation sans aucun apport en azote (i), un apport d’azote strictement limité à la période amont au débourrement (ii), un apport permanent en azote (iii). Les résultats montrent que le peuplier peut absorber de l’azote avant le débourrement et que son absorption produit des effets significatifs. Elle conduit en particulier à une forte poussée racinaire qui permet d’augmenter la teneur en eau des tissus caulinaires. Selon les essais, l’effet de l’absorption d’azote avant débourrement et sur le débourrement lui-même dépend de la structure de la plante, de la structure des bourgeons et de la température de conservation hivernale pour le traitement de la dormance. D’autre part, l’application d’azote avant débourrement améliore significativement la croissance des nouvelles pousses en augmentant, avec un temps de croissance identique dans les essais, la surface foliaire et la matière sèche. L’absorption d’azote pendant la transition de croissance maintient en partie la teneur en azote des tissus des plantes et en améliore même la teneur dans les jeunes racines et les jeunes pousses. Elle influence aussi l’utilisation des réserves carbonées. Les résultats de cette étude montrent que l’azote appliqué au printemps et avant débourrement joue un rôle significatif sur la physiologie du jeune arbre et sa reprise de croissance au printemps. / Nitrogen status widely impacts tree physiological process. However, most research concentrated on endogenous carbon and nitrogen remobilization in spring neglected the hypothesis of significant effect of nitrogen uptake before bud break on nitrogen and carbohydrate reserve used, and the quality of new growth. This study undertook experimental-based research on young poplar (Populus tremula × Populus alba, clone INRA 717-1B4) with different structures. Three series of experiments were conducted in a coordinated manner : the one year old scion (i), the young stump (ii), and the system then reiterated associated the “stump”, and the “scion” (iii). The experiments were to study plants in a controlled environment with soilless culture and three terms of nitrogen supply: without nitrogen supply (i), with a limited supply prior to bud break (ii), and with continuous nitrogen supply (iii). Results show that poplar can uptake nitrate before bud break and it found to have significant effect by induce a strong root pressure which in turn increased water content of all tissues. Accordingly, the effect of nitrogen uptake before bud break on bud break time depended on plant architecture, bud structure, and temperature during winter to break down bud dormancy. In addition, nitrogen uptake before bud break had significantly effect on the growth and development of new shoots after bud break by increase leaf area and dry weight of new shoots. It also influenced on the quantity of nitrogen and non-structural carbon reserves in all tissues especially increase nitrogen contents in roots and new shoots. Therefore, results indicate nitrogen supply before bud beak plays a significant role on plant physiology and quality of the re-growth.

Although mango (Mangifera indica L.) has been studied for many years, numerous problems still elude researchers. The objectives of the current trials were to study the effects of some cultural practices (fruit thinning, panicle/ bud/ renewal/ post-harvest pruning) and chemicals (Corasil.E, potassium nitrate/urea, paclobutrazol) on various vegetative, floral, yield and quality parameters. The study meant to address problems of both South African and Ethiopian mango growers. The thinning (on ‘Sensation’) and pruning (on ‘Tommy Atkins’ and ‘Keitt’) experiments were conducted for two seasons (2001-2003) at Bavaria Fruit Estate in South Africa. Effects of Paclobutrazol and Potassium nitrate were studied on ‘Tommy Atkins’ during 2002-2003 season at Upper Awash Agro-industry Enterprise in Ethiopia. Complementary effects of paclobutrazol and potassium nitrate on floral induction were studied in growth chamber experiments at the experimental farm of University of Pretoria on ‘Tommy Atkins’ and ‘Keitt’ mango. Where fruit on ‘Sensation’ were thinned to one and two fruit per panicle, a significant increase was obtained for most of the fruit quantitative parameters. The treatments where one fruit and two fruit per panicle were retained and 50% panicles removed, produced a significant increase in size of the fruit, fruit qualitative parameters and fruit retention percentage. Corasil.E produced very small sized fruit with a considerable percentage of “mules” (fruit without seed). Trees subjected to severe thinning intensities showed earlier revival of starch reserves and better vegetative growth. Applications of paclobutrazol (1- (4-chlorophenyl) –4,4-dimethyl-2- (1,2,4- triazol-1-yl) pentan-3-ol) at rates of 5.50 and 8.25 g a.i. per tree, both as a soil drench and spray applications, on ‘Tommy Atkins’ mango were effective in suppressing vegetative growth as compared to the control. Consequently, the trees from these treatments had higher total non-structural carbohydrate in their shoots before flowering which led to higher results of percentages of shoots flowering, number of panicles produced, percentages of hermaphrodite flowers, yield and quality of the fruit. Trees that received panicle pruning (during full bloom) treatment at the point of apical bud attachment, were observed to be induced for synchronized re-flowering and also attained more fruit per panicle. On the other hand, trees on which renewal pruning (early in the season) and post-harvest pruning (especially for early cultivars) treatments were applied, have been observed for the development of an adequate number of productive inflorescences. Post-harvest pruning treatments also resulted in greater vegetative growth on both cultivars. The responses to pruning treatments were greater especially in ‘Tommy Atkins’ than ‘Keitt’. The trend for the interaction of duration and chemicals in Tommy Atkins and Keitt mango cultivars revealed the possible floral induction complementary effect of PBZ after the trees were induced only for 15 days at 10/15 oC temperature. Higher potassium nitrate concentrations especially in combination with urea (5 litre solution of 4% KNO3+0.5 g urea tree-1 and 5 litres of 4% KNO3+1 g urea tree-1 ) produced higher results for most of the flowering and yield parameters in ‘Tommy Atkins’. / Thesis (DPhil (Horticulture))--University of Pretoria, 2006. / Plant Production and Soil Science / unrestricted

Denna studie kommer att utforska hur dolda bud påverkar tilliten till budgivningsprocessen, där syftet med studien är att undersöka påverkan på förtroendet hos köpare, säljare och potentiella köpare på bostadsmarknaden. Med denna undersökning vill vi ta reda på vilka utmaningar dolda bud skapar för att tillföra kunskap och lärdomar för mäklarbranschen. I arbetet diskuteras det hur dolda bud har bidragit till en opålitlig budgivningsprocess, där de inblandade aktörerna ofta misstänker att mäklaren har undanhållit information. Detta sker trots att Fastighetsmäklarlagen inte innefattar en reglerad budgivning ännu, där säljaren i nuläget avgör om budgivningen ska hållas öppen eller vara stängd. Forskningsstudiens teoretiska ramverk består av tillit, risk och osäkerhet, transparens och asymmetrisk information. Påverkan på transaktionskostnader och det marknadsmässiga perspektivet behandlas även i det teoretiska ramverket. Arbetet baseras på en kvalitativ undersökning med semistrukturerad intervjuteknik som består av nio genomförda intervjuer. Dessa baseras på deltagare som tidigare varit i kontakt med dolda bud i en försäljningsprocess. Resultatet av studien visar att det finns en stor okunskap om fenomenet dolda bud vilket i sin tur skapar missförstånd och misstro. Slutligen visar forskningen på att det finns en rad faktorer som påverkas negativt av de dolda buden med ett fåtal skillnader mellan de inblandade aktörerna. De främsta slutsatser som framhävs i forskningsrapporten är förtroendets påverkan hos de olika parterna, då ett flertal vann medan andra förlorade i budgivningsprocesserna. / This study will investigate how hidden bidding affects trust in the bidding process where the aim of the study is to investigate the impact on the trust of buyers, sellers and potential buyers in the housing market. With this survey, we want to find out what challenges hidden bids create in order to add knowledge and lessons for the brokerage industry. The work discusses how hidden bids have contributed to an unreliable bidding process where the parties involved often suspect that the broker has withheld information. This is despite the fact that the Real Estate Agents Act does not yet include regulated bidding, where the seller currently decides whether the bidding should be kept open or closed. The research study's theoretical starting points consist of trust, risk and uncertainty, transparency, and asymmetric information. The impact on transaction costs and the market perspective is also addressed in the theoretical framework. The work is based on a qualitative survey with a semi-structured interview technique consisting of nine completed interviews. These are based on participants who have previously been in contact with hidden bids in a sales process. The results of the study show that there is a great deal of ignorance about the phenomenon of hidden bids, which creates misunderstandings and mistrust. Finally, the study shows that there are several factors that are negatively affected by the hidden bids with a few differences between the parties involved. The main conclusions that are highlighted are the impact of trust on the part of the various parties, as several won while others lost in the bidding processes.

Taste buds are chemosensory endorgans embedded in the oral epithelium composed of cells that undergo continuous replacement. Mature taste cells live on average 10-14 days and are replaced by new cells when they die. However, the mechanism by which taste cells are produced and integrated into the taste bud as mature taste cells remains unknown. Previous studies approached this issue from either cell cycle gene expression properties or lineage tracing of precursor cells. In our study, we apply a new fate mapping technique that combines these two ideas. This technique, Mosaic Analysis with Double Markers, allows for simultaneous gene knockout and subsequent tracking of single cells. This allows us to study the potency of precursor cells supplying the taste bud while analyzing how gene function regulates the maturation pathway these taste cells take. The following experiments illustrate the initial phase of this investigation.