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THE ASSOCIATION BETWEEN POLYPLOIDY AND CLONALITY IN THE HERBACEOUS PLANT, CHAMERION ANGUSTIFOLIUM (ONAGRACEAE)Baldwin, Sarah J 14 May 2012 (has links)
The co-occurrence of polyploidy and clonal reproduction among plant species has long been recognized, but the evolutionary mechanisms underlying the association are unknown. Here, I investigate whether polyploidy increases the magnitude of clonality, either directly or indirectly, by comparing the extent and spatial structure of clones between diploid and tetraploid Chamerion angustifolium in a greenhouse environment and natural populations. In the greenhouse, tetraploid plants allocated 90.4% more dry mass to root buds, the primary mechanism of clonal reproduction, than diploids. Per unit root mass, tetraploids produced 44% fewer root buds and the average position of the root buds along the root was 47% closer to the stem than in diploids. In natural populations, the magnitude of clonality in tetraploid C. angustifolium was similar or less than in diploids. However, clones were spatially aggregated in all diploid populations but only in two of five tetraploid populations. Average clone patch diameter, however, was not significantly different between diploids (3.9 m) and tetraploids (2.5 m). These data do not support the hypothesis that clonality increases as a result of genome duplication. Rather, it is possible that clonality is linked to genome duplication because clonal diploids are predisposed for polyploid formation and establishment. / National Science and Engineering Research Council, Canada Research Chair Program, Canadian Foundation for Innovation
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Caracterização das combinações de copa e portaenxerto de pereira europeia quanto a ocorrência de entomosporiose e ao abortamento de gemas florais no Estado de Santa Catarina / Characterization of combinations of cup and rootstok of european pear cultivars as a entomosporiose leaf spot and indexes the necrosis in flower bud abortiom in Santa Catarina StateGonçalves, Mayra Juline 09 February 2012 (has links)
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Previous issue date: 2012-02-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study aimed to evaluate the temporal dynamics of Entomosporiose leaf spot in diferente combinatos of European pear cultivars (Rocha, Abate fetel and Santa Maria) on diferente rootstocks (Quince A and Quince Adams) and evaluate the etiology and development of necrosis in flower bud in diferente combinations European pear (Clapp s Favorite, Packham s Triumph, Conference, William s, Rocha, Abate, Fetel, Santa Maria, Decana) European pear cultivars on diferente rootstocks (Quince A, Quince C and Quince Adams) at conditions of the Santa atarina state. In Entomosporiose the incidence and severity were assessed weekly to onset of symptoms in 100 random leaves, distributed in 4 medium branches per plant. The incidence was calculated by the percentage of leaves with at least one lesion relative to the total number of leaves evaluated and severity of infection classes. The experiments were consucted in comercial orchards in the municipalities of Fraiburgo and Urupema/Santa Catarine State, during the growing seasons 2009/10 and 2010/11. During the period from June to September of each growing season, 10 buds of each pear combination with rootstock at the phenological stages of vegetative rest and opening the scales, were evaluated for the presence of necrosis. The buds were incubated in BDA and King B culture media for fungi and bactéria identification, respectively. The fungal identification was performed by visualization of reproductive structures and identification Keys. For bacterial identification, purê cultures have been featured by LOPAT as described by Shaad (2001). The necrosis presence or not were correlated with pears combinations phenological stage and causal agente of necrosis. The Entomosporiose incidence and severity date were used to plotted the progress curves of the disease and the epidemix compared with respect to: a) the beginning of symptoms appearance (BSA); b) time to reach maximum incidence and severity of disease (TRMDI and TRMDS), c) masimum incidence and severity (Smax and Imax) and d) área under teh cuirve of progress in the incidence and severity of disease (AUCPID and AUCPSD). The necrotic buds showed the presence of Pseunomonas syringae and Xanthomonas spp bactéria and there is no significant difference of the correlation of buds necorsis occurrences with cultivars, rootstock and phonological stage evaluated. The identified bactéria were involved, in necrosis of flowers buds of the European pear as a factor inducing seconfary. All combinations of cultivars with rootstocks were susceptible to E. mespili, and the higher intensity of disease occurred in the Santa Maria cultivar, regardless of the rootstock used. There were significant diferences in the Entomosporiose incidence and severity progress curves of the cultivars Rocha, Abate Fetel and Santa Maria / Este trabalho objetivou avaliar a dinâmica temporal da Entomosporiose em diferentes combinações de cultivares copa pereira europeia (Rocha, Abate Fetel e Santa Maria) sobre diferentes portaenxertos (Marmelo A e Marmelo Adams) e avaliar a etiologia e a evolução da necrose de gemas florais em combinações de cultivares copa (Clapp´sFavorite, Packham´sTriumph, Conference, William s, Rocha, Abate Fetel, Santa Maria, Decana) de pereira europeia sobre diferentes portaenxertos (Marmelo A, Marmelo C e Marmelo Adams) nas condições edafoclimáticas do estado de Santa Catarina. Na Entomosporiose a incidência e a severidade foram avaliadas semanalmente ao surgimento dos primeiros sintomas em 100 folhas aleatórias, distribuídas em 4 ramos medianos por planta. A incidência foi calculada pela percentagem das folhas com pelo menos uma lesão em relação ao número total de folhas avaliadas e a severidade através de classes de infecção. Os experimentos de necrose foram conduzidos em pomares comerciais nos municípios de Fraiburgo e Urupema durante as safras 2009/10 e 2010/11. No período de junho a setembro foram coletas 10 gemas de cada combinação e área experimental, nos estádios fenológicos de repouso vegetativo e abertura das escamas, e avaliadas quanto a presença de necrose. As gemas foram incubadas em meios de cultura específico para fungos (BDA) e para bactérias (B de King). A identificação fúngica foi realizada através da visualização das estruturas reprodutivas e chaves de identificação. Para identificação bacteriana, culturas puras foram caraterizadas através do LOPAT segundo a descrição de Shaad (2001). A presença ou não de necrose de gemas foram correlacionadas com as combinações de cultivares e portaenxertos, estádio fenológicos e agente causal. Com os dados de incidência e severidade da Entomosporiose foram plotadas curvas de progresso da doença e as epidemias comparadas em relação a: a) início do aparecimento dos sintomas (IAS); b) tempo para atingir a máxima incidência e severidade da doença (TAMID e TAMSD); c) valor máximo de incidência e severidade (Imax e Smax) e d) área abaixo da curva do progresso da incidência e da severidade da doença (AACPID e AACPSD). Nas gemas necrosadas, verificou-se a presença das bactérias Pseudomonas syringae do gênero e Xanthomonas e não houve diferenças significativas na ocorrência de necrose das gemas tanto nas combinações de cultivares e portaenxerto quanto nos estádios fenológicos avaliados. As bactérias identificadas estão envolvidas na necrose de gemas florais de pereira europeia como um fator indutor secundário. Todas as combinações de cultivares e portaenxertos foram suscetíveis a Entomosporium mespili e a maior intensidade de doença ocorreu na cultivar Santa Maria, independente do portaenxerto utilizado. Houve diferença significativa nas curvas de progresso da doença tanto da incidência quanto da severidade da Entomosporiose nas cultivares Rocha, Abate Fetel e Santa Maria
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Podridão floral dos citros: histopatologia de Colletotrichum acutatum / Postbloom fruit drop: histopatology of Colletotrichum acutatumJoão Paulo Rodrigues Marques 09 August 2012 (has links)
A podridão floral dos citros (PFC) é uma doença causada pelo fungo Colletotrichum acutatum responsável por causar grandes danos à produção de citros no Brasil. A doença surge apenas em botões florais com 8mm de comprimento ou maiores, levando a formação de lesões alaranjadas nas pétalas, lesões necróticas no estigma, promove a queda prematura dos frutos e a retenção do cálice e pedúnculo, sendo este último sintoma denominado estrelinha. Este trabalho tem por objetivo: observar o modo de penetração do fungo no hospedeiro Citrus sinensis Valência e os estágios posteriores da colonização, verificar se há fatores estruturais e químicos pré-formados que expliquem o porquê do fungo não conseguir infectar botões florais com menos de 8mm, caracterizar anatomicamente o sintoma estrelinha e estigmas lesionados, investigar ultraestruturalmente pétalas inoculadas, analisar se há o estabelecimento de uma infecção quiescente nos tecidos foliares, analisar grãos de pólen após a inoculação in vivo e in vitro com o fungo. Botões florais sadios, pétalas e estigmas com e sem lesões, foram submetidos às técnicas convencionais de microscopia de luz e eletrônica. Folhas e grãos de pólen foram inoculados e analisados. Foi desenvolvida uma nova técnica de coloração para tecidos vegetais infectados por fungos. A resistência dos botões florais menores que 8mm pode estar associada às barreiras químicas e estruturais pré-formadas. O ápice, nesses botões, apresenta papilas entremeadas, cristais de oxalato de cálcio no mesofilo e câmara subestomática e cavidades de óleo localizadas muito próximas umas das outras. Botões com 8mm e 12mm possuem, no ápice, papilas com arranjo frouxo, ausência de cristais e maior distanciamento entre as cavidades de óleo. No ápice da pétala, verificou-se que as células papilosas são osmóforos. No sintoma estrelinha, nota-se sob a região de abscisão do ovário a instalação de um meristema de cicatrização. A lignificação das paredes das células da medula do receptáculo e do pedúnculo floral está associada à retenção destas estruturas na planta. Nas pétalas infectadas, o C. acutatum pode penetrar intra, intercelularmente e via estômato. O fungo pode crescer de modo subcuticular e intramural e coloniza todos os tecidos da pétala. A nova técnica de coloração se mostrou muito útil nas análises histopatológicas. O fungo associa-se aos tecidos vasculares. Acérvulos ocorrem em ambas as faces das pétalas. A cutícula nos estágios mais avançados da lesão apresenta-se alterada, ou seja, ocorre a perda da ornamentação estriada e maior deposição de material lipofílico. A síntese de materiais lipofílicos envolve o retículo endoplasmático liso e rugoso e plastídios. Vesículas provenientes de dictiossomos e de corpos multivesiculares são observadas ao longo da parede celular e estão associadas ao depósito de material lipofílico na cutícula. No estigma lesionado há a formação de uma camada de proteção. O fungo apresenta quimiotropismo e cresce em direção aos grãos de pólen infectando-os 24 horas após a inoculação. Sugere-se que C. acutatum pode utilizar grãos de pólen para a sua dispersão. Após 48 horas da inoculação as folhas apresentam conídios germinados com apressórios. / The postbloom fruit drop (PFD) is a disease caused by Colletotrichum acutatum responsible for causing great damage to citrus crops in Brazil. The disease appears only in flower buds 8 mm in length or greater, leading to orange lesions in petals, necrotic lesions on the stigma, promoting the young fruit drop and the retention of the calyx and peduncle, which is called buttons. In this context, this study aimed to: observe the fungus penetration mode into the host Citrus sinensis \'Valência\' and the later stages of colonization; study the presence of preformed structural and chemical factors to explain why the fungus cannot infect floral buds with less than 8 mm in length; characterize anatomically the symptom \"buttons\" and injured stigmas; investigate the ultrastructural changes in tissues of inoculated petals; analyze whether there is the establishment of a quiescent infection in leaf tissues, analyze pollen grains after inoculation in vivo and in vitro with the fungus. Healthy buds, petals and stigmas with and without lesions, were processed and analyzed using conventional light and electron microscopy techniques. Leaves and pollen grains were also inoculated and analyzed with light microscopy. It was developed a new staining method for fungal-infected plant tissues. The resistance of flower buds smaller than 8mm may be associated with preformed structural and chemical barriers. These buttons display the apex with interspersed papillae, with crystals in mesophyll and substomatic chamber and oil cavities, which are located very close to each other on the abaxial surface. In 8mm and 12mm flower buds, the papilas in the apex become weakly interspaced, the crystals are not observed and there is the increase of the distance between the oil cavities. The papillose cells are osmophores. In the symptom \"button\", it is noted in the abscission of the ovary, an installation of wound meristem. There is also the lignification in the pith of receptacle and pedicel that can be associated with the retention of these structures in the plant. In infected petals, it was found that C. acutatum can penetrate intra and intercelullar or via stomata. The fungus may grow subcuticular and intramural and colonize all tissues of petal. The new staining technique developed has proved very useful for histopathological analysis. The fungus is closely associated with vascular tissues. The acervulli occur on both surfaces of petals. The cuticle in the later stages of the lesion is altered, i.e., there is loss of striated ornamentation and increased deposition of lipophilic material. The synthesis of lipophilic materials involves rough and smooth endoplasmic reticulum and plastids. Vesicles from dictyosomes and multivesicular bodies were observed throughout the cell wall and are associated with the deposit of lipophilic material in the cuticle. There is the formation of protective layer over the stigma damaged area. The fungus shows chemotropism and grows toward the pollen infecting it 24 hours after inoculation. It is suggested that C. acutatum can use pollen grains for dispersal. After 48 hours of inoculation, the leaves have germinated conidia with appressoria.
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Organogênese in vitro em laranja azeda (Citrus aurantium L.) e transformação genética de limão \'Cravo\' (Citrus limonia L. Osbeck) e laranja \'Valência\' (Citrus sinensis L. Osbeck) com o gene da replicase do Marafivirus / In vitro organogenesis in sour orange (Citrus aurantium L.) and genetic transformation of Rangpur lime (Citrus limonia L. Osbeck) and Valencia sweet orange (Citrus sinensis L. Osbeck) with the Marafivirus replicase geneRosely Pereira da Silva 30 June 2008 (has links)
Embora desfrute de inegável importância econômica, os citros estão sujeitos a muitos problemas sanitários sendo alguns, limitantes para o cultivo como é o caso das doenças causadas por vírus. A morte súbita dos citros é uma doença relacionada à combinação copa/ porta-enxerto e manifesta sintomas na região da enxertia sobre porta-enxertos intolerantes. Embora sua etiologia não tenha sido determinada, há indicações que a causa da MSC esteja relacionada a uma estirpe do vírus da tristeza dos citros (CTV), a um vírus do gênero Marafivirus, ou a uma associação entre eles. Uma vez que a transformação genética têm sido considerada como uma ferramenta auxiliar a programas de melhoramento de citros, o objetivo deste trabalho foi obter plantas transgênicas de limão \'Cravo\' e laranja \'Valência\' contendo o gene da replicase do Marafivirus e estudar a regeneração e obtenção de plantas in vitro de laranja azeda, via organogênese, visando futuros trabalhos de transformação genética. Experimentos para indução da organogênese in vitro foram realizados avaliando-se citocininas (BAP, TDZ e CIN), em diferentes concentrações, isoladamente ou em combinação com ANA, condições de luminosidade (fotoperíodo de 16 h e escuro por 30 dias), meios de cultivo e explantes (provenientes de plantas germinadas in vitro e de plantas mantidas em estufa). Além disso, avaliou-se o enraizamento dos brotos regenerados. Para a transformação genética, explantes de limão \'Cravo\' e laranja \'Valência\' foram inoculados e co-cultivados com a estirpe EHA 105 de Agrobacterium tumefaciens contendo o gene da replicase do Marafivirus (em seqüência sense e antisense interligadas por um íntron). A construção gênica foi elaborada a partir do plasmídeo pCAMBIA 2201, dirigidas pelo promotor 35S e terminador NOS, contendo ainda o gene de seleção nptII. A transformação foi confirmada por análises de PCR e \'Southern blot\'. A transcrição do gene foi avaliada por RT-PCR e \'northern blot\'. A adição de BAP, combinada ou não com ANA, e em combinações com CIN ao meio de cultivo, assegurou maior formação de gemas adventícias em segmentos de epicótilo de laranja azeda. Entretanto, TDZ não se mostrou favorável a essa resposta, que também é afetada pela ausência de luz. Os explantes provenientes do cultivo in vitro mostraram-se mais favoráveis à resposta organogênica. O enraizamento das brotações de laranja azeda regeneradas foi obtido no meio MT com metade da concentração de sais, sem ou com auxinas. Foi possível obter plantas transgênicas de limão \'Cravo\' e de laranja \'Valência\' contendo o gene da replicase do Marafivirus utilizando-se segmentos internodais como explantes. A análise de \'Southern blot\' confirmou a integração de um a quatro eventos de inserção do transgene no genoma das plantas. A transcrição do gene da replicase do Marafivirus e do gene nptII foi observada por RT-PCR. / In spite of great economic importance, the citrus industry is affected by many phytopathological problems some, limiting its cultivation such as virus-caused diseases. The citrus sudden death disease is related to scion/rootstock combinations and manifests symptoms in the grafting area of intolerant rootstocks. Although its etiology has not been determined, there are indications that the cause of MSC might be related to a strain of the Citrus tristeza virus (CTV), to a virus of the Marafivirus group, or to an association of both viruses. Since the genetic transformation has been considered as an auxiliary tool to programs of citrus improvement, the objectives of this work were to obtain transgenic plants of the Rangpur lime and Valencia sweet orange containing the Marafivirus replicase gene and study the in vitro regeneration of sour orange plants through organogenesis, aiming for future work in genetic transformation. Experiments for induction of in vitro organogenesis were carried out evaluating citocinins (BAP, TDZ and KIN), in different concentrations, separately or in combination with NAA, lighting conditions (photoperiod of 16 hours and darkness for 30 days), cultivation media and explants (coming from in vitro germinated plants and from green house cultivated plants). Besides this, rooting of the regenerated shoots was evaluated. For the genetic transformation, Rangpur lime and Valencia sweet orange explants were inoculated and co-cultivated with the EHA-105 Agrobacterium tumefaciens strain containing the Marafivirus replicase gene (in sense and antisense sequence linked by an intron). The genetic construct used derived from the pCAMBIA 2201 plasmid, driven by the 35S promoter and NOS terminator, containing the selection nptII gene. The genetic transformation was confirmed by PCR and Southern blot analysis. The gene transcription was evaluated by RT-PCR and northern blot. The addition of BAP to the culture medium, combined or not with NAA, and in combinations with KIN, assured a greater formation of adventitious buds in sour orange epicotyl segments. However, TDZ was not favorable to this response, that is also affected by the absence of light. Explants coming from in vitro cultivation were more favorable to the organogenic response. Rooting of sour orange regenerated shoots was obtained in MT medium with half the salt concentration, with or without auxin. It was possible to obtain transgenic Rangpur lime and Valencia sweet orange plants containing the Marafivirus replicase gene using internodal segments as explants. The Southern blot analysis confirmed the integration of one to four copies of the transgene in the plant genome. The transcription of the Marafivirus replicase gene and the nptII gene was observed by RT-PCR.
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Desenvolvimento de metodologia radiográfica e volumétrica dos diferentes estágios de desenvolvimento dentário para qualificação de material biológico em Engenharia Tecidual / Development of radiographic and volumetric metodologies from diferentes tooth development stages as a qualification for harvesting biological material for Tissue EngeneeringDuailibi Neto, Eduardo Felippe 12 March 2013 (has links)
A utilização de Células-tronco e técnicas da Engenharia Tecidual representa um grande avanço tecnológico e beneficiará muitos pacientes com suas conquistas. A descoberta de germes dentários como uma fonte confiável de células-tronco possibilitou diversas pesquisas nesta área. Duailibi et al. (2011) sugeriram uma nova classificação de desenvolvimento dentário baseada pela quantidade de material biológico coletado indicando a necessidade de métodos de diagnóstico por imagem para esta nova classificação. Na literatura diversos trabalhos indicam métodos de classificação dentária e métodos para estimar a idade fisiológica de indivíduos. O presente estudo tem o objetivo de adaptar alguns destes métodos para estimar o estágio de desenvolvimento proposto por Duailibi et al. (2011) consequentemente indicando a quantidade de células-tronco esperadas nas amostras. Para tanto, submeteu-se uma coleção de 67 dentes previamente classificados por Duailibi et al. (2011) à técnica rpcl e à tcfc para a obtenção de imagens e a aplicação de técnicas de estimativas por proporções lineares e volumétricas. Os resultados por análises lineares indicaram valores de R2 para o método de proporção de comprimento CDCP de 0,14050; CCCP de 0,65369; CCCR de 0,5408; CDCR de 0,54074; o método de proporção de área APAD de 0,23925; e método de proporção de volume VPVD de 0,08553, com valor de p menor ou igual à 0,05. Concluindo este estudo indica-se o método de rpcl utilizando a análise do comprimento entre coroa e polpa como o mais indicado para estimar o estágio de desenvolvimento. / The usage of human dental stem cells and tissue engineering technics represents a huge tecnological development and it may benefits many patients in a promissing future. The discovery of suitable source of human dental stem cells were made using tooth buds. Duailibi et al. 2011 indicated a new tooth classification on a stem cell harvesting based research, sugesting new methods for diagnosis these stages. Several method were developed for dental age assesement. The presente study aims to evaluate some of these dental age technics and make adaptations for estimating Duailibi et al. 2011 tooth stages. A 67 tooth sample previoulsy classificated by Duailbi et al. 2011 were submited through periapical parallel long cone X-rays and CBCT analysis. Age estimation ratio methods were applied by measuring tooth/root lenth, crown/root lenth, tooth/pulp lenth, crown/pulp lenth, tooth/poulp área and tooth/pulp volume. Results indicated by linear regression analisys a R2 value of tooth/pulp lenth 0,14050; crown/pulp lenth 0,65369; crown/root lenth 0,5408; tooth/root lenth 0,54074; pulp/tooth volume 0,23925; e tooth/pulp volume de 0,08553, with p value of 0,005. In conclusion , the best method for estimating Duailibi et al. 2011 tooth classification techinic is made by using periapical long cone X-rays using crown/pulp lenth ratio.
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Kryokonservierung und in vitro Kultur von Pyrus pyraster (L.) BURGSD. und Sorbus torminalis (L.) CRANTZKadolsky, Marianne 27 August 2007 (has links)
Um die Erhaltung und Nutzung der beiden gefährdeten, hochwertigen Baumarten Pyrus pyraster und Sorbus torminalis zu fördern, wurden Methoden der in vitro Kultur und der Kryokonservierung evaluiert und optimiert. Kryokonservierung von Sorbus torminalis wird erstmalig beschrieben. Als Explantatquellen dienten Winterknospen, überwiegend von selektierten Bäumen aus Nachkommenschaftsprüfungen. Von den geprüften Faktoren Nährmedienzusammensetzung, Kulturgefäß, Schneidetechnik, Temperatur, Pikiertermin und physiologischer Zustand der Explantate erwies sich letzterer als entscheidend. Die Reiser mit den Winterknospen wurden in zwei aufeinander folgenden Jahren im November, Dezember, Januar, Februar und März geerntet und sowohl direkt als auch nach 2-monatiger Lagerung bei +5°C bearbeitet. Bei Pyrus führte Ernte im März zu den besten Ergebnissen der in vitro Kultur. Die besten Ergebnisse der Kryokonservierung wurden von Ernten im November bis Januar erzielt. Bei Sorbus differierten die Ergebnisse der in vitro Kultur zwischen den Jahren so stark, dass keine Aussage möglich war. Bei der Kryokonservierung waren die Erntetermine Dezember und Januar erfolgreich. Zur Kryokonservierung von in vitro Material wurden die Alginat-Einkapselung und die Droplet-Technik geprüft. Mit Pyrus wurden mit der Droplet-Technik Vitalitätsraten zwischen 7 und 13% erzielt, wenn eine bisher unveröffentlichte Kombination von Vorbehandlungen und PVS2-Vitrifizierung eingesetzt wurden. Die Kryokonservierung wirkte sich im Vergleich weder positiv noch negativ auf Wachstum und Entwicklung aus. Die Diskrepanz zwischen der Nutzung der tiefen Dormanz der Knospen und des damit verbundenen natürlichen Frostschutzes für die Kryokonservierung einerseits und der Vermeidung von tiefer Dormanz bei Etablierung von in vitro Kulturen wegen der damit verbundenen Wachstumshemmung andererseits wird diskutiert. / To facilitate conservation and utilization of the endangered valuable tree species Pyrus pyraster and Sorbus torminalis methods of in vitro culture and cryopreservation were evaluated and optimized. Cryopreservation of Sorbus torminalis is described for the first time. The explant sources were dormant winter buds, mainly from selected trees from progeny trials. The evaluation of media composition, culture vessel, cutting technique, date of pricking and physiological state proved the latter to be crucial. Twigs with winter buds were harvested in two subsequent years in November, December, January, February and March and were processed immediately and after two months of storage at +5°C. With Pyrus, the best results in in vitro culture were obtained after harvest in March while for cryopreservation harvest in November to January proved to be best. With Sorbus, the results in in vitro culture differed between the years and no conclusion could be drawn. Cryopreservation was successful after harvest in December and January. Encapsulation in alginate and the droplet-technique were evaluated for the cryopreservation of in vitro material. Vitality rates of 7 to 13% were achieved with Pyrus using the droplet-technique with a combination of pre-treatments and PVS2-vitrification not published so far. In comparison, cryopreservation was of neither positive nor negative influence on growth and development. The discrepancy between deep dormancy of buds being useful for cryopreservation because of its natural frost protection and the need to avoid it because it impedes in vitro culture is discussed.
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Desenvolvimento de metodologia radiográfica e volumétrica dos diferentes estágios de desenvolvimento dentário para qualificação de material biológico em Engenharia Tecidual / Development of radiographic and volumetric metodologies from diferentes tooth development stages as a qualification for harvesting biological material for Tissue EngeneeringEduardo Felippe Duailibi Neto 12 March 2013 (has links)
A utilização de Células-tronco e técnicas da Engenharia Tecidual representa um grande avanço tecnológico e beneficiará muitos pacientes com suas conquistas. A descoberta de germes dentários como uma fonte confiável de células-tronco possibilitou diversas pesquisas nesta área. Duailibi et al. (2011) sugeriram uma nova classificação de desenvolvimento dentário baseada pela quantidade de material biológico coletado indicando a necessidade de métodos de diagnóstico por imagem para esta nova classificação. Na literatura diversos trabalhos indicam métodos de classificação dentária e métodos para estimar a idade fisiológica de indivíduos. O presente estudo tem o objetivo de adaptar alguns destes métodos para estimar o estágio de desenvolvimento proposto por Duailibi et al. (2011) consequentemente indicando a quantidade de células-tronco esperadas nas amostras. Para tanto, submeteu-se uma coleção de 67 dentes previamente classificados por Duailibi et al. (2011) à técnica rpcl e à tcfc para a obtenção de imagens e a aplicação de técnicas de estimativas por proporções lineares e volumétricas. Os resultados por análises lineares indicaram valores de R2 para o método de proporção de comprimento CDCP de 0,14050; CCCP de 0,65369; CCCR de 0,5408; CDCR de 0,54074; o método de proporção de área APAD de 0,23925; e método de proporção de volume VPVD de 0,08553, com valor de p menor ou igual à 0,05. Concluindo este estudo indica-se o método de rpcl utilizando a análise do comprimento entre coroa e polpa como o mais indicado para estimar o estágio de desenvolvimento. / The usage of human dental stem cells and tissue engineering technics represents a huge tecnological development and it may benefits many patients in a promissing future. The discovery of suitable source of human dental stem cells were made using tooth buds. Duailibi et al. 2011 indicated a new tooth classification on a stem cell harvesting based research, sugesting new methods for diagnosis these stages. Several method were developed for dental age assesement. The presente study aims to evaluate some of these dental age technics and make adaptations for estimating Duailibi et al. 2011 tooth stages. A 67 tooth sample previoulsy classificated by Duailbi et al. 2011 were submited through periapical parallel long cone X-rays and CBCT analysis. Age estimation ratio methods were applied by measuring tooth/root lenth, crown/root lenth, tooth/pulp lenth, crown/pulp lenth, tooth/poulp área and tooth/pulp volume. Results indicated by linear regression analisys a R2 value of tooth/pulp lenth 0,14050; crown/pulp lenth 0,65369; crown/root lenth 0,5408; tooth/root lenth 0,54074; pulp/tooth volume 0,23925; e tooth/pulp volume de 0,08553, with p value of 0,005. In conclusion , the best method for estimating Duailibi et al. 2011 tooth classification techinic is made by using periapical long cone X-rays using crown/pulp lenth ratio.
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Functional Anatomy and Development of Cactus RamificationsSchwager, Hannes 12 November 2015 (has links) (PDF)
Cacti (Cactaceae) represent a family of highly specialized angiosperm plants with a native range of distribution restricted to the American continents. Columnar cacti of the sub-family Cactoideae evolved in adaptation to their arid or semi-arid habitats characteristics that distinguish them from most other dicot plants, e.g. the stem succulence with a strongly vascularized storage parenchyma and the presence of the spine wearing areoles.
Although cacti have been in cultivation since the discovery of America, some studies even suggest the agricultural use in pre-colombian times, and many scientific investigations were carried out on the functional morphology and anatomy with regard to biomechanical adaptations of the found structures, no research focused on the branch-stem attachment.
The most conspicuous features of such a ramification are the pronounced constrictions at the branch-stem junctions that are also present in the lignified vascular structures within the succulent cortex. Based on Finite Element Analyses of ramification models it could be demonstrated that these indentations in the region of high flexural and torsional stresses are not regions of structural weakness, e.g. allowing vegetative propagation. On the contrary, they can be regarded as anatomical adaptations to increase the stability by fine-tuning the stress state and stress directions in the junction along prevalent fiber directions.
The development of the woody support structure within the succulent cortex of the parental shoot can be traced back to the leaf and bud traces of the dormant axillary buds. Surprisingly, these initials also develop into another woody structure supporting the flowers of the cacti. As these two support structures differ significantly in their macroscopic and microscopic anatomy and as they develop from the same initial state as leaf/bud traces, another objective of this work was to analyze the secondary growth of the two structures with traditional botanic investigation methods.
The results of these investigations reveal a wood dimorphism consisting of an early parenchymatous phase followed later by fibrous wood in both kind of support structure. In vegetative branches, the woody support structures have the typical ringlike arrangement as found in the stele of the parental shoot, whereas the flower support structures have a reticular arrangement of interconnected woody strands. This fundamentally different anatomy of the support structures results from the formation of an interfascicular cambium between the leaf/bud traces when a vegetative branch forms or its absence in the case of a flower.
After shedding light on the functional morphology and anatomy of the cactus ramification and their development the question arises if the found load adaptation strategies may serve to improve technical fiber composite structures analogue to the design recommendation developed from the biomechanical analyses of tree ramifications. Such a biomimetic transfer from the cactus ramification as biological role model to a technical implementation and the adaptation of the fine-tuned geometric shape and arrangement of lignified strengthening tissues might contribute to the development of alternative concepts for branched fiber-reinforced composite structures within a limited design space.
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Ekologická omezení odnožování z kořenů u mokřadních rostlin / Ecological constraints limiting the root-sprouting ability in wetland plant speciesSOSNOVÁ, Monika January 2010 (has links)
Wetland plant species rely largely on vegetative reproduction. Although all types of clonal growth organs are found in wetlands, special adaptations, e.g., turions, fragmentation and budding, are more frequent in true aquatic communities. However, root-sprouting is underrepresented, although it can be beneficial under disturbed conditions. This thesis focuses on ecological constrains potentially hindering root-sprouting in wetlands. This ability was studied in a wetland herb Rorippa palustris in relation to life history, injury timing and carbon economy of a plant. In addition, plant regeneration following submergence and severe disturbance was assessed.
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Functional Anatomy and Development of Cactus RamificationsSchwager, Hannes 09 July 2015 (has links)
Cacti (Cactaceae) represent a family of highly specialized angiosperm plants with a native range of distribution restricted to the American continents. Columnar cacti of the sub-family Cactoideae evolved in adaptation to their arid or semi-arid habitats characteristics that distinguish them from most other dicot plants, e.g. the stem succulence with a strongly vascularized storage parenchyma and the presence of the spine wearing areoles.
Although cacti have been in cultivation since the discovery of America, some studies even suggest the agricultural use in pre-colombian times, and many scientific investigations were carried out on the functional morphology and anatomy with regard to biomechanical adaptations of the found structures, no research focused on the branch-stem attachment.
The most conspicuous features of such a ramification are the pronounced constrictions at the branch-stem junctions that are also present in the lignified vascular structures within the succulent cortex. Based on Finite Element Analyses of ramification models it could be demonstrated that these indentations in the region of high flexural and torsional stresses are not regions of structural weakness, e.g. allowing vegetative propagation. On the contrary, they can be regarded as anatomical adaptations to increase the stability by fine-tuning the stress state and stress directions in the junction along prevalent fiber directions.
The development of the woody support structure within the succulent cortex of the parental shoot can be traced back to the leaf and bud traces of the dormant axillary buds. Surprisingly, these initials also develop into another woody structure supporting the flowers of the cacti. As these two support structures differ significantly in their macroscopic and microscopic anatomy and as they develop from the same initial state as leaf/bud traces, another objective of this work was to analyze the secondary growth of the two structures with traditional botanic investigation methods.
The results of these investigations reveal a wood dimorphism consisting of an early parenchymatous phase followed later by fibrous wood in both kind of support structure. In vegetative branches, the woody support structures have the typical ringlike arrangement as found in the stele of the parental shoot, whereas the flower support structures have a reticular arrangement of interconnected woody strands. This fundamentally different anatomy of the support structures results from the formation of an interfascicular cambium between the leaf/bud traces when a vegetative branch forms or its absence in the case of a flower.
After shedding light on the functional morphology and anatomy of the cactus ramification and their development the question arises if the found load adaptation strategies may serve to improve technical fiber composite structures analogue to the design recommendation developed from the biomechanical analyses of tree ramifications. Such a biomimetic transfer from the cactus ramification as biological role model to a technical implementation and the adaptation of the fine-tuned geometric shape and arrangement of lignified strengthening tissues might contribute to the development of alternative concepts for branched fiber-reinforced composite structures within a limited design space.
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