O Eucalyptus cloeziana se destaca pelas propriedades tecnológicas da sua madeira, principalmente em razão da sua durabilidade, densidade e resistência. Entretanto, essa espécie apresenta limitações quanto ao enraizamento adventício de estacas, dificultando a obtenção de mudas clonais e o avanço nos programa de melhoramento da espécie. Nesta perspectiva, o presente trabalho teve por objetivo avaliar a micropropagação de E. cloeziana a partir de materiais juvenis e adultos como uma técnica para a propagação da espécie. Para tanto, o trabalho foi dividido em quatro partes. No primeiro estudo, estabeleceu-se um protocolo de organogênese indireta de E. cloeziana a partir de hipocótilos e cotilédones. No segundo estudo avaliou-se o resgate de matrizes adultas de E. cloeziana através da indução de brotações epicórmicas em megaestacas. No terceiro estudo definiu-se um protocolo de micropropagação via proliferação de gemas axilares de matrizes adultas de E. cloeziana. Por último, no quarto estudo avaliou-se o rejuvenescimento in vitro de matrizes adultas de E. cloeziana por meio da micropropagação via proliferação de gemas axilares e a microestaquia, a partir das matrizes rejuvenescidas in vitro,para comprovar a viabilidade do cultivo in vitro como alternativa de propagação da espécie. A organogênese indireta de E. cloeziana mostrou-se possível e dependente do tipo de explante, dos reguladores de crescimento, suas concentrações e combinações utilizadas nas diferentes fases de morfogênese. As brotações adventícias foram micropropagadas e aclimatizadas com sucesso, obtendo-se mudas clonais de E. cloeziana. No resgate das matrizes adultas de E. cloeziana ocorreu maior indução de brotações epicórmicas para as megaestacas coletadas na época do ano com maior pluviosidade e temperatura. O diâmetro médio das megaestacas, compreendido entre 2,0 e 5,0 cm, correspondeu ao mais adequado para a obtenção de maior número de brotações epicórmicas no resgate das matrizes adultas de E. cloeziana. O protocolo de micropropagação das matrizes adultas de E. cloeziana foi estabelecido com sucesso, utilizando como explantes, brotações epicórmicas induzidas nas megaestacas. A multiplicação in vitro das microcepas foi realizada com êxito no meio de cultura WPM suplementado com BAP e ANA, cujas concentrações variaram entre os genótipos. Os tratamentos de \"pulse\" com GA3 não foi adequado para promover o alongamento in vitro das brotações, o qual foi obtido coma redução da concentração do BAP (0,1 mg L-1). A aclimatização e o enraizamento ex vitro das microestacas foi realizada com sucesso nos miniestufas, garantindo a obtenção de mudas clonais das matrizes adultas de E. cloeziana para a composição de um microjardim clonal. As microcepas das árvores matrizes de E. cloeziana apresentaram diferentes taxas de multiplicação in vitro, apresentando especificidades dos materiais genéticos à micropropagação. Os resultados da microestaquia das matrizes rejuvenescidas in vitro de E. cloeziana corroboraram as evidências de rejuvenescimento desses genótipos durante a micropropagação. Além disso, constatou-se que outros fatores, além da maturidade, podem estar atuando diretamente na recalcitrância do E. cloeziana ao enraizamento adventício. Dessa forma, comprovou-se a importância da micropropagação como uma ferramenta para a clonagem de E. cloeziana, abrindo perspectivas para investigações futuras para otimizar os métodos de propagação da espécie. / Eucalyptus cloeziana has importance for its wood technologic characteristics, mainly the durability, density and resistance. Moreover, this species has limitations on adventitious rooting of cuttings, having difficulties to obtain clonal seedlings and to advance in the improvement programs. In this perspective, the present work aimed to evaluate the E. cloeziana micropropagation by juvenile and mature explants as a technique for propagation this specie. Therefore, the work was divided into four basic studies. The first study was the establishment of the protocol to indirect organogenesis to E. cloeziana hypocotyls and cotyledons. In the second study evaluated the rescue of E. cloeziana adult matrices for epicormic shoots induction in crow branches. In the third study was definite a micropropagation protocol by axillary branching of E. cloeziana adult matrices. Finally, in the fourth study was evaluated the in vitro rejuvenation of E. cloeziana adult matrices to micropropagation by axillary branching and the micro-cutting technique of in vitro rejuvenated matrices to prove the in vitro culture viability to the propagation of this specie. The indirect organogenesis E. cloeziana was dependent of explant type and growth regulator and its concentration used in the different morphogenesis phases. The adventitious shoots were multiplicities and acclimatized with success to obtain E. cloeziana clonal seedlings. The vegetative rescue results of E. cloeziana adult matrices showed a higher induction epicormic shoots from branches collected in warmer and rain season. The medium diameter between 2.0 and 5.0 cm was considered the better to obtain higher epicormic shoots number in vegetative rescue. The micropropagation protocol of E. cloeziana adult matrices was realized with success using epicormic shoots, induced in the branches, like explants. The shoots in vitro multiplication was realized on WPM medium culture, supplemented with BA and NAA. The better growth regulators concentrations were different for each genotype. Treatments pulse with GA3 was not suitable to promote the elongation of shoots in vitro, which was obtained by the BA concentration reduction at 0.1 mg L-1. The in vitro shoots acclimatization and ex vitro rooting was performed in mini-incubators with success and it permit to obtain clonal seedlings from E. cloeziana adult matrices to form a clonal micro-garden. The shoots of E. cloeziana adult matrices showed different in vitro multiplication rates with specific-genotype responses to micropropagation. The micro-cutting technique results of E. cloeziana adult matrices provide evidence that occurred in vitro rejuvenation of these genotypes, during the micropropagation. Therefore, the nursery results showed that other factors, not only maturation, can be involved in the adventitious recalcitrance of E. cloeziana. In conclusion, the importance micropropagation like a biotechnological tool to propagation of E. cloeziana was proved and opened important perspectives for future investigations to optimize the propagation methods for this specie.

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O Greening ou Huanglongbing (HLB) é a doença mais devastadora dos citros na atualidade. Levantamentos mostram que a enfermidade está presente em cerca de 60% dos talhões do parque citrícola paulista, demostrando sua grande capacidade de disseminação. O psilídeo-asiático-dos-citros, Diaphorina citri Kuwayama (Hemiptera: Psylidae) é um importante vetor dos agentes causais associados ao HLB. Sua capacidade de dispersão está intimamente relacionada ao progresso da doença no campo. Não se tem conhecimento dos principais fatores que possam interferir no deslocamento do inseto até plantas cítricas. Sendo assim, a presente pesquisa teve como objetivo investigar a influência de alguns fatores ambientais sobre a dispersão do psilídeo D. citri, discutindo-os em relação à epidemiologia do HLB dos citros. Inicialmente foram realizados testes em laboratório para adaptação da metodologia de marcação, liberação e recaptura usando pó fluorescente. Grupos de psilídeos adultos foram marcados e sua sobrevivência avaliada, verificando a influência de quatro cores distintas de pó fluorescente (amarelo, azul, rosa e verde). Posteriormente realizaram-se estudos de campo com quatro liberações de psilídeos, com marcadores distintos, em quatro áreas experimentais circulares e adjacentes, sendo formadas por círculos concêntricos localizados a 18, 24 e 30 m de cada centro de liberação. Os insetos foram monitorados visualmente e por armadilhas adesivas amarelas. Nas 1ª e 2ª liberações, foi verificado o efeito das coberturas vegetais milho (Zea mays L.) e capim-braquiária (Brachiaria decumbens Stapf.) na dispersão de D. citri. Já nas 3ª e 4ª liberações, foi estudado o efeito de brotações no deslocamento do inseto. A influência dos ventos foi avaliada em ambas as situações. A análise de sobrevivência indicou que não houve interferência das quatro cores de pó fluorescente em D. citri, sendo a marcação visível no corpo dos insetos durante os 48 dias de avaliação. De acordo com as análises fatoriais, não foi observada qualquer interferência da cobertura vegetal milho, medindo 2 m de altura, na dispersão do inseto. Na presença de brotações, adultos de D. citri dispersaram menos em busca de novos hospedeiros, acumulando-se nas primeiras linhas de plantas cítricas. Após 6 horas da liberação, a capacidade de dispersão máxima de D. citri foi três vezes maior na condição de ausência de brotações. A dispersão média e a área total abrangida variaram assimptoticamente com o tempo, ou seja, o inseto possui inicialmente grande velocidade de dispersão, porém esta tende a se estabilizar com o incremento do tempo. Esses resultados deixam claro que D. citri realiza movimentos dependendo das condições fenológicas de seus hospedeiros, ressaltando a grande importância de se intensificar métodos de controle regionais em épocas em que o fluxo vegetativo das plantas cítricas não é tão intenso. / Greening or Huanglongbing (HLB) is the most devastating disease of citrus today. Surveys show that the disease is present in about 60% of the park stands citrus Sao Paulo, demonstrating its great ability to spread. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psylidae) is an important vector of the causative agents associated with HLB. Its ability to spread is closely related to disease progression in the field. There is no knowledge of the main factors that can affect the movement of the insect to citrus plants. Thus, the present study aimed to investigate the influence of some environmental factors on the dispersal of psyllid D. citri, discussing them in relation to the epidemiology of HLB of citrus. Initially tests were performed in the laboratory to adapt the methodology of marking, release and recapture using fluorescent powder. Groups of adult psyllids were marked and their survival assessed by checking the influence of four different colors of fluorescent powder (yellow, blue, pink and green). Subsequent studies were conducted with four field releases psyllids, with distinct markers on four experimental areas adjacent circular and being formed by concentric circles located at 18, 24 and 30 m for each center release. The insects were monitored visually and by yellow sticky traps. In the 1st and 2nd releases, was to determine the effect of cover crops maize (Zea mays L.) and grass (Brachiaria decumbens Stapf.) in the dispersal of D. citri. In the 3rd and 4th releases, were studied the effect of shootings in insect movement. The influence of the wind has been evaluated in both situations. Survival analysis indicated that there was no interference of the four colors of fluorescent powder on D. citri, and marking was visible on the body of insects during the 48 days of evaluation. According to the factor analysis, no interference of vegetation corn (measuring 2 m in height) was observed in the dispersal of the insect. In the presence of shoots, adult D. citri less dispersed looking for new hosts and it accumulate in the first lines of citrus plants. After 6 hours of release, the D. citri maximum dispersal was three times higher in the condition of absence of shoots. The dispersal medium and the total area covered varied asymptotically with time, ie the insect has initially high speed dispersal, but this tends to stabilize with increasing time. These results clearly show that D. citri performs movements depending on the phenology of its hosts, emphasizing the importance of intensifying regional control methods at times when the flow of plant citrus is not intense.

Made available in DSpace on 2016-12-23T14:37:37Z (GMT). No. of bitstreams: 1 Fabiola Lacerda de Souza Barros.pdf: 6472406 bytes, checksum: d8013423c68ed8608b3a5668f18e35ac (MD5) Previous issue date: 2008-07-10 / Foram realizados experimentos in vivo e in vitro objetivando a propagação assexuada do mamoeiro (Carica papaya L.). O experimento in vivo foi realizado com a cultivar Golden , do grupo Solo , em lavoura comercial com dois anos e meio de idade em final de produção, onde se estudou o efeito da poda em diferentes alturas do tronco (inteira sem o ápice, 0,5, 1,0, 1,5, 2,0 m) a partir do solo, para se identificar qual destas proporcionaria maior rendimento de brotações laterais aptas à estaquia. Uma contagem de brotações aos 25 dias após a poda a diferentes alturas foi realizada no experimento in vivo. As brotações contabilizadas foram podadas e, posteriormente, feita outra contagem de novas brotações aos 50 dias. Com a poda a 2,0 metros de altura, obteve-se maior número de brotações aos 25 e aos 50 dias, no período do verão, mostrando viabilidade no reaproveitamento de lavouras em final de produção na propagação assexuada. Meios de enraizamento foram testados nos experimentos in vitro. No primeiro experimento foram estudadas as concentrações do meio MS a 100%, 50% da força e MS modificado associados aos níveis do regulador de crescimento AIB (0; 0,1; 0,2; 0,4 e 0,8 mg L-1). No primeiro experimento o enraizamento foi insatisfatório devido à contaminação bacteriana, aliada a uma possível fitotoxidez do antibiótico utilizado (cefotaxima 100 mg L-1), ou ainda, ao acúmulo de citocinina remanescente da fase de multiplicação, prejudicando as reações morfogênicas e o enraizamento. No segundo experimento, utilizou-se meio de enraizamento contendo ou não carvão ativo, na concentração 1gL-1, associado aos níveis do regulador de crescimento ANA (0; 0,25; 0,5; 1,0; 2,0 mgL-1). Também não se obteve sucesso no enraizamento, provavelmente pela quantidade ou associação dos antibióticos utilizados (rifampicina 300 mg L-1 +cloranfenicol 100 mg L-1) ao meio de cultura, mesmo tendo controlado acontaminação bacteriana. O carvão ativo a 1 g L-1 acrescentado ao meio, apesar de não auxiliar o enraizamento, mostra significância de forma isolada, sendo benéfico ao desenvolvimento da parte aérea e impedindo o calejamento na base dos ramos, mostrando eficiência na adsorção de substâncias tóxicas do meio de cultivo, em relação ao tratamento que não recebeu o carvão ativo / The experiments were carried out in vivo and in vitro having of the objective the assexual reproduction of the papaya tree (Carica papaya L.). The experiment in vivo was accomplished with to cultivate Golden, of the group Sole, in commercial farming with two and a half years of age in production end, where it was studied the effect of the pruning in different heights of the trunk (it completes without the apex, 0,5, 1,0, 1,5, 2,0 m) starting from the ground, and to identify which of these it would provide larger income of capable production of lateral shoots to the cutting. With the pruning to 2,0 meters of height was obtained larger shoots number, in two consecutive pruning, in other words, to the 25 and 50 days, in the period of the summer, showing viability in the recycling of farmings in production end in the assexual reproduction. In the experiments in vitro were tested rooting medium. In the first experiment were studied the concentrations of the MS medium of 100%, 50% of the force and MS modified associates at the levels of the growth regulator AIB (0; 0,1; 0,2; 0,4 and 0,8 mg L-1). The rooting was unsatisfactory due to the bacterial contamination, allied to a possible phytotoxity of the used antibiotic (cephotaxin 100 mg L-1), or still, to the kitocinin accumulation remaining of the multiplication phase harming the morphogenics reactions and the rooting. In the second experiment was used rooting medium containing (1 g L-1) or not active coal, associate at the levels of the growth regulator ANA (0; 0,25; 0,5; 1,0 and 2,0 mg L-1). It was not also obtained success in the rooting probably for the amount or association of the antibiotics used (Rifampicin 300 mg L-1 + cloranphenicol 100 mg L-1) in the culture medium, even having controlled the bacterial contamination. The active coal to 1 g L-1 increased to the medium, in spite of not aiding the rooting show significance in an isolated way, being beneficial to the development of the aerial part and impeding the calluse in the base of the shoots, showing efficiency in the adsorption of poisonous substances of the culture medium, in relation to the treatment that didnt receive the active coal

O Eucalyptus cloeziana se destaca pelas propriedades tecnológicas da sua madeira, principalmente em razão da sua durabilidade, densidade e resistência. Entretanto, essa espécie apresenta limitações quanto ao enraizamento adventício de estacas, dificultando a obtenção de mudas clonais e o avanço nos programa de melhoramento da espécie. Nesta perspectiva, o presente trabalho teve por objetivo avaliar a micropropagação de E. cloeziana a partir de materiais juvenis e adultos como uma técnica para a propagação da espécie. Para tanto, o trabalho foi dividido em quatro partes. No primeiro estudo, estabeleceu-se um protocolo de organogênese indireta de E. cloeziana a partir de hipocótilos e cotilédones. No segundo estudo avaliou-se o resgate de matrizes adultas de E. cloeziana através da indução de brotações epicórmicas em megaestacas. No terceiro estudo definiu-se um protocolo de micropropagação via proliferação de gemas axilares de matrizes adultas de E. cloeziana. Por último, no quarto estudo avaliou-se o rejuvenescimento in vitro de matrizes adultas de E. cloeziana por meio da micropropagação via proliferação de gemas axilares e a microestaquia, a partir das matrizes rejuvenescidas in vitro,para comprovar a viabilidade do cultivo in vitro como alternativa de propagação da espécie. A organogênese indireta de E. cloeziana mostrou-se possível e dependente do tipo de explante, dos reguladores de crescimento, suas concentrações e combinações utilizadas nas diferentes fases de morfogênese. As brotações adventícias foram micropropagadas e aclimatizadas com sucesso, obtendo-se mudas clonais de E. cloeziana. No resgate das matrizes adultas de E. cloeziana ocorreu maior indução de brotações epicórmicas para as megaestacas coletadas na época do ano com maior pluviosidade e temperatura. O diâmetro médio das megaestacas, compreendido entre 2,0 e 5,0 cm, correspondeu ao mais adequado para a obtenção de maior número de brotações epicórmicas no resgate das matrizes adultas de E. cloeziana. O protocolo de micropropagação das matrizes adultas de E. cloeziana foi estabelecido com sucesso, utilizando como explantes, brotações epicórmicas induzidas nas megaestacas. A multiplicação in vitro das microcepas foi realizada com êxito no meio de cultura WPM suplementado com BAP e ANA, cujas concentrações variaram entre os genótipos. Os tratamentos de \"pulse\" com GA3 não foi adequado para promover o alongamento in vitro das brotações, o qual foi obtido coma redução da concentração do BAP (0,1 mg L-1). A aclimatização e o enraizamento ex vitro das microestacas foi realizada com sucesso nos miniestufas, garantindo a obtenção de mudas clonais das matrizes adultas de E. cloeziana para a composição de um microjardim clonal. As microcepas das árvores matrizes de E. cloeziana apresentaram diferentes taxas de multiplicação in vitro, apresentando especificidades dos materiais genéticos à micropropagação. Os resultados da microestaquia das matrizes rejuvenescidas in vitro de E. cloeziana corroboraram as evidências de rejuvenescimento desses genótipos durante a micropropagação. Além disso, constatou-se que outros fatores, além da maturidade, podem estar atuando diretamente na recalcitrância do E. cloeziana ao enraizamento adventício. Dessa forma, comprovou-se a importância da micropropagação como uma ferramenta para a clonagem de E. cloeziana, abrindo perspectivas para investigações futuras para otimizar os métodos de propagação da espécie. / Eucalyptus cloeziana has importance for its wood technologic characteristics, mainly the durability, density and resistance. Moreover, this species has limitations on adventitious rooting of cuttings, having difficulties to obtain clonal seedlings and to advance in the improvement programs. In this perspective, the present work aimed to evaluate the E. cloeziana micropropagation by juvenile and mature explants as a technique for propagation this specie. Therefore, the work was divided into four basic studies. The first study was the establishment of the protocol to indirect organogenesis to E. cloeziana hypocotyls and cotyledons. In the second study evaluated the rescue of E. cloeziana adult matrices for epicormic shoots induction in crow branches. In the third study was definite a micropropagation protocol by axillary branching of E. cloeziana adult matrices. Finally, in the fourth study was evaluated the in vitro rejuvenation of E. cloeziana adult matrices to micropropagation by axillary branching and the micro-cutting technique of in vitro rejuvenated matrices to prove the in vitro culture viability to the propagation of this specie. The indirect organogenesis E. cloeziana was dependent of explant type and growth regulator and its concentration used in the different morphogenesis phases. The adventitious shoots were multiplicities and acclimatized with success to obtain E. cloeziana clonal seedlings. The vegetative rescue results of E. cloeziana adult matrices showed a higher induction epicormic shoots from branches collected in warmer and rain season. The medium diameter between 2.0 and 5.0 cm was considered the better to obtain higher epicormic shoots number in vegetative rescue. The micropropagation protocol of E. cloeziana adult matrices was realized with success using epicormic shoots, induced in the branches, like explants. The shoots in vitro multiplication was realized on WPM medium culture, supplemented with BA and NAA. The better growth regulators concentrations were different for each genotype. Treatments pulse with GA3 was not suitable to promote the elongation of shoots in vitro, which was obtained by the BA concentration reduction at 0.1 mg L-1. The in vitro shoots acclimatization and ex vitro rooting was performed in mini-incubators with success and it permit to obtain clonal seedlings from E. cloeziana adult matrices to form a clonal micro-garden. The shoots of E. cloeziana adult matrices showed different in vitro multiplication rates with specific-genotype responses to micropropagation. The micro-cutting technique results of E. cloeziana adult matrices provide evidence that occurred in vitro rejuvenation of these genotypes, during the micropropagation. Therefore, the nursery results showed that other factors, not only maturation, can be involved in the adventitious recalcitrance of E. cloeziana. In conclusion, the importance micropropagation like a biotechnological tool to propagation of E. cloeziana was proved and opened important perspectives for future investigations to optimize the propagation methods for this specie.

Die vorliegende Arbeit befasst sich mit den morphologischen bzw. cytologischen Anomalien eines mutierten Klons 5/74/2 aus der haploiden Sorte `Kleiner Liebling` von Pelargonium zonale. An histologischen Schnitten wurden die Ploidiestufen verschiedener Zellschichten von Cytochimären durch Chromosomenzählung direkt bestimmt. Es wurde festgestellt, dass die Epidermis in den Blättern vom Klon 5/74/2 verschiedene Ploidiestufen aufweist. Solche Variabilität trat in den L2- und L3-bürtigen Zellschichten nicht auf. Somit wurde hier erstmals eine Periklinalcytochimäre mit verschiedenen Ploidie in der Epidermis nachgewiesen. Es zeigte aber zwei unterschiedliche Zustände im Sprossscheiteln: Beim ersten sind die Zellen der L1 Polyploidie. Beim zweiten sind die Zellen der L1 haploid. Die Polyploidisierung der Epidermis setzt sich bei dem Wachstum des Blattes fort. Bei den cytologischen Untersuchungen wurden anomale Mitosen mit irregulären Chromsomenanordnungen (multipolare) und mehrkernige Zellen in der Epidermis beobachtet. Als weitere Anomalie traten eingeschnürte Zellkerne in den Epidermiszellen auf. Die Pflanzen mit stark behaarten und gekräuselten Blättern vom Klon 5/74/2 waren nicht stabil. Die Epidermis des gekräuselten und stark behaarten Blattes enthielt wie die des stark behaarten Blattes oder die des gekräuselten Blattes verschiedene Ploidiestufe. Die Epidermiszellen der meisten Pflanzen mit normal glatten Blättern lagen im diploiden Bereich. Die morphologische Kräuselung und Behaarung hing mit dem Auftreten der somatischen Variabilität in Epidermis von Klon 5/74/2 zusammen. Adventivpflanzen mit Hilfe eines modifizierten Mediums aus Kallus von Pelargonium zonale sind in in vitro-Kultur zu regenerieren. Dabei zeigte die Zugabe von Perlit und anschließend PVP die beste Wirkung gegen die Bildung von Polyphenolverbindungen im Pelargonium-Gewebe. TDZ ist einer der effektivsten Wuchsstoffe für die Kallusinduktion und Regeneration der Adventivsprosse bei Pelargonium. Die Blatt- und die Blütenfarbe der Adventivpflanzen aus dem dreifach markierten chimärischen `Weißer Liebling` (WRR, GWG, DHH) stimmten mit dem Genotyp der L1-Zellen überein. Die Stomatalängen der Adventivpflanzen aus dem Klon 5/74/2 (PHH) lagen wie die von `Weißer Liebling` (DHH) meistens im Bereich von diploid, nur selten von tetraploid oder gemischten. Die ursprünglich polyploiden und haploiden Zellen der kultivierten Gewebe traten nach der Regeneration nicht mehr auf. Dies weist auf eine Zellmutation und -selektion während der in vitro-Kultur hin. Aus chimärischen Geweben der GWG- und GGW-Varianten von `Mrs. Pollock` wurden 8,6% Adventivsprosse mit neuen chimärischen Blattmustern regeneriert. Das Ergebnis zeigt, dass die Zellen mindestens in L2 und L3-bürtigen Gewebeschichten der Explantaten zur Regeneration der Adventivsprossen teilgenommen haben. / A mutated clone with different ploidy levels in epidermis cells, 5/74/2, of the haploid `Kleiner Liebling` of Pelargonium zonale was investigated to answer the question that how the different ploidy levels were generated. Such a variability did not appear in L2- and L3-derived cells. Consequently, clone 5/74/2 is a periclinal cytochimera with a mixed ploidy epidermis. This type of cytochimera with different ploidy levels in epidermis has not been reported up to now. The epidermis of the blistered leaf or of the hairy leaf is polyploid with different ploidy levels, like the epidermis of the shoots with blistered and hairy leaves. Epidermis cells of normal shoots are diploid. The morphologically blistered leaf surface seems to be the result of a somatic variability in epidermis. Histological investigations of clone 5/74/2 showed two different ways of development of the somatic variability: the cells of the L1 in the apical meristem were already polyploid and the cells in the apical meristem were haploid. Cytological analyses revealed anomalies of the mitosis in epidermis cells of clone 5/74/2. Various irregular chromosomal arrangements (multipolar mitosis) and polynuclear cells were observed. Additionally, nuclei with constrictions were found. Adventitious shoots were successfully regenerated from callus of adult tissue of Pelargonium in vitro on modified medium. The combination of Perlit and PVP showed the best effects against the formation of phenolic substances on Pelargonium tissues during in vitro-culture. TDZ was the most effective substance for callus induction and regeneration of adventitious shoots. The colour of leaves and flowers in the adventitious plants regenerated from tissue of the triply marked chimera ’Weißer Liebling’ (GWG, WRR, DHH) were correlated with the genotype of L1. However, somaclonal variability of ploidy levels was observed. The size of stomata on adventitious plants from the clone 5/74/2 (PHH) was similar to from DHH-tissue, mostly in the range of diploid, only rarely in tetraploid or mixed ploidy. Derivates of the original polyploid and haploid cells of the cultivated tissue were not found. Evidently, cell selection and mutation occurred during the in vitro-culture. 8.6% Adventitious shoots with new chimeral patterns were regenerated from chimeral GWG- and GGW-explants of Pelargonium zonale `Ms. Pollock`. More chimeras from GGW explants were obtained than from GWG explants. This result showed that cells in both L2 and L3 derived tissues of the explants were involved in the regeneration of the adventitious shoots.

In this study, the immediate and short term (three months) effects of conservation burning have been investigated in coniferous forests in southeastern Sweden. Five tree species were investigated Picea abies (Norway spruce), Pinus sylvestris (Scots pine), Populus tremula (aspen), Betula pendula (silver birch) and Betula pubescens (downy birch), as well as ground vegetation of mosses, dwarf shrubs and ground lichens. Burning increased the proportion of live deciduous tree shoots from 51 % to 81 % and the live tree shoot size distribution of four tree species was significantly changed by fire. Fire affected the tree species differently. Three months after burning deciduous tree species exhibited strong sprouting, while P. sylvestris had established many seedlings, significantly increasing its share of the tree stand. P. abies lacked any visible positive response to burning and its number of live shoots decreased by 83 %. Mineral soil was exposed on only the moss vegetation and covered just 4 % of the studied plot area. The initial vegetation response to fire was negative, but significant dwarf shrub recovery was detected three months after burning.

Ce travail de doctorat traite l’optimisation des méthodes d’extraction des composés phénoliques à partir des sous-produits de la viticulture et de la viniculture, à savoir les sarments de vigne et les marcs de raisins. Plusieurs technologies innovantes sont appliquées et comparées : l’extraction accélérée par solvant (EAS), les décharges électriques de haute-tension (DEHT), les ultrasons (US) et les champs électriques pulsés (CEP). Les extractions solide-liquide faites sur les sarments ont montré que, parmi les solvants étudiés, l’eau est le moins efficace. L’ajout de la β-cyclodextrine dans l’eau améliore le procédé d’extraction mais est moins efficace que les mélanges hydroéthanoliques. L’extraction en milieu alcalin donne le meilleur rendement en composés phénoliques. L’intensification de l’extraction des composés phénoliques des sarments est possible grâce aux nouvelles technologies d’extraction. L’efficacité des méthodes testées est la moindre avec les US, moyenne avec les CEP pour atteindre le meilleur rendement phénolique avec les DEHT. La filtrabilité de ces extraits est d’autant plus lente que leur composition est complexe. L’ultrafiltration membranaire permet une très bonne purification et concentration des composés phénoliques. L’étude des mécanismes d’action des DEHT a permis d’identifier les phénomènes favorisant l’extraction des composés phénoliques à partir des sarments. Un effet mécanique des DEHT, capable de fragmenter les sarments, est en majorité responsable de cette amélioration. Le procédé énergivore du broyage pourra alors être omis. Un effet électrique contribuant également à l’intensification du procédé d’extraction est démontré. La formation de peroxyde d’hydrogène durant le traitement par DEHT est quantifiée mais ne semble pas altérer les composés phénoliques qui sont des molécules à capacité antiradicalaire élevée. Quant aux études portées sur les marcs de raisins, la variation simultanée de plusieurs paramètres opératoires a permis l’optimisation de l’extraction aqueuse et hydroéthanolique des composés phénoliques en ayant recours à la méthodologie de surface de réponse (MSR). Le passage d’un milieu aqueux à un milieu hydroéthanolique a permis d’améliorer nettement le procédé d’extraction solide-liquide des composés phénoliques et l’utilisation de l’EAS a permis l’augmentation du rendement en composés phénoliques jusqu’à trois fois par rapport à l’optimum obtenu en milieu hydroéthanolique. / This study deals with the optimization of the extraction methods of phenolic compounds from viticulture and viniculture by-products, namely vine shoots and grape pomace. Several innovative technologies were tested and compared: high voltage electrical discharges (HVED), accelerated solvent extraction (ASE), ultrasounds (US) and pulsed electric fields (PEF). The solid-liquid extraction conducted on vine shoots showed that, amongst the studied solvents, water is the least effective. The addition of the β-cyclodextrin to water improves the extraction process but remains less effective than that with hydroethanolic mixtures. The extraction in alkaline medium gives the highest phenolic compound extraction yields. The intensification of phenolic compound extraction from vine shoots was possible thanks to new extraction technologies. The effectiveness of the tested methods was the least with US, followed by PEF to accomplish the highest phenolic yield with HVED. The filterability of the extracts was slower when their composition was complex, and the membrane technology allowed a good purification and concentration of phenolic compounds. The reason behind the high effectiveness of HVED was investigated. The action mechanisms of HVED were studied in details. A mechanical effect of HVED provoked vine shoots fragmentation and particle size reduction. This was the main phenomenon responsible for the intensification of the extraction process. It also suggested that a grinding pretreatment would not be necessary prior to HVED, which considerably diminishes the energy input of the overall process. The presence of a non-mechanical effect and its contribution in the efficiency of HVED were also shown. The formation of hydrogen peroxide during the treatment was observed. However it did not seem to alter vine shoot phenolic compounds since these demonstrated a high radical scavenging capacity. As for the studies conducted on grape pomace, the simultaneous variation of several operating parameters allowed the aqueous and hydroethanolic optimization of phenolic compound extraction from these byproducts by response surface methodology (RSM). The passage from an aqueous to a hydroethanolic medium clearly improved the solid-liquid extraction of phenolic compounds from grape pomace. The use of ASE further increased the phenolic compound yield up to three times as compared to the optimum obtained with a hydroethanolic solvent.

The study design includes the new building of the Moravian wine and cultural center on the Hlinky street in Brno. There are spaces designed for wineries (producing, maturing and storing wine), administration, congress center (with business lounges), spa, restaurant and gastro-studio, shops, exhibition, hotel and parking. The building is trapezoidal shape and replace the existing object in a vacant lot, only some historical cellars will be retained. The supporting structure consists of reinforced concrete skeleton system and reinforced concrete reinforcing inner core around the staircase leading to all floors, the building is covered by a flat green roof. On the facade dominates the element that its shape resembles the vine shoots, which holds the glass facade. The construction is divided into two expansion units - the main part of the building and garage, which is set into the ground sloping towards the south. The building has six floors and one underground floor. The first four floors are connected by open gallery at the southern facade.