O gênero Passiflora é um excelente modelo para estudos de transição de fase, pois há diferenças morfológicas evidentes entre as plantas nas fases juvenil, adulta vegetativa e adulta reprodutiva. Há relatos de que quando se introduzem ápices de plantas adultas de Passiflora edulis, quer estejam na fase vegetativa ou reprodutiva, em condições de cultivo in vitro, observa-se a regressão à fase juvenil. O objetivo geral deste trabalho foi caracterizar as modificações morfológicas envolvidas na reversão à juvenilidade de ápices caulinares adultos de P. edulis quando cultivados em meio de cultura sem adição de reguladores de crescimento além de avaliar o efeito de reguladores de crescimento no intuito de evitar a reversão à juvenilidade no cultivo in vitro de P. edulis. O trabalho foi desenvolvido a partir da combinação da técnica da micropropagação in vitro e microscopia eletrônica de varredura, sendo dividido em três experimentos: o cultivo in vitro de ápices caulinares em meio MSM sem reguladores de crescimento e análise de amostras em microscópio eletrônico de varredura, cultivo in vitro de ápices caulinares em meio MSM com reguladores de crescimento e cultivo in vitro de ápices caulinares em meio MSM adicionados com BAP na concentração de 10?M com análises de amostras em microscópio estereoscópico. Através de micrografias eletrônicas e imagens estereoscópicas, foi possível caracterizar as fases do desenvolvimento de P. edulis, demonstrando os caracteres morfológicos assim como a manutenção das estruturas cultivadas in vitro. Não foi possível observar o rejuvenescimento in vitro quando ápices caulinares foram cultivados em meio ½ MS ou MSM sem adição de reguladores de crescimento. O rejuvenescimento de ápices caulinares adultos de P. edulis foi obtido a partir do cultivo em meio MSM com adição de 10?M de BAP. Entretanto, o rejuvenescimento foi parcial, pois nem todas as características foram revertidas. A formação de folhas lanceoladas in vitro e o desenvolvimento de brotos adventícios in vitro caracterizaram a reversão à juvenilidade em P. edulis. O meio MSM suplementado com 10 ?M de BAP foi favorável ao desenvolvimento de gemas axilares, além de facilitar a indução do rejuvenescimento dos ápices caulinares. Logo, a reversão à juvenilidade foi alcançada sob as condições in vitro adotadas / The genus Passiflora is an excellent model for phase transition studies because there are obvious morphological differences between plants in the juvenile, adult vegetative and adult reproductive phases. It has been reported that when introducing shoot tips of adult plants of Passiflora edulis, whether in the vegetative or reproductive phase, under in vitro culture conditions, regression to the juvenile phase is observed. The general objective of this work was to characterize the morphological changes involved in the reversion to juvenility of adult shoot tips of P. edulis when grown in culture medium without addition of growth regulators in addition to evaluating the effect of growth regulators in order to avoid reversion to juvenility in the in vitro cultivation of P. edulis. The work was developed from the combination of in vitro micropropagation and scanning electron microscopy, and was divided in three experiments: in vitro culture of shoot tips in MSM medium without growth regulators and analysis of samples in scanning electron microscope; in vitro culture of shoot tips in MSM medium with growth regulators and in vitro culture of shoot tips in MSM medium added with BAP at the concentration of 10?M and analysis of samples under a stereoscopic microscope. Through electronic micrographs and stereoscopic images, it was possible characterize the phases of the development of P. edulis, demonstrating the morphological traits as well as the maintenance of the structures cultivated in vitro. In vitro rejuvenation could not be observed when shoot tips were grown in ½ MS or MSM medium without addition of growth regulators. The rejuvenation of P. edulis adult shoot tips was obtained from the cultivation in MSM medium with the addition of 10?M BA. However, the rejuvenation was partial, since not all the characteristics were reversed. The formation of in vitro lanceolated leaves and the development of in vitro adventitious shoots characterized the reversion to juvenility in P. edulis. The MSM medium supplemented with 10?M BA was favorable to the development of axillary buds, in addition to facilitating the induction of the rejuvenation of the shoot tips. Therefore, the reversion to juvenility was achieved under the in vitro conditions adopted

O nitrogênio e o potássio são essenciais para o desenvolvimento das plantas, são os nutrientes extraídos em altas quantidades e sua disponibilidade pode alterar as características produtivas e nutricionais das plantas. Assim, objetivou-se estudar: a) as características morfogênicas e a produção de massa seca da parte aérea e das raízes da Brachiaria brizantha cv. Marandu e b) a concentração de nitrogênio, potássio, nitrato e amônio nas lâminas de folhas recém-expandidas e nas raízes, bem como o valor SPAD, a atividade da enzima redutase do nitrato, a distribuição percentual do nitrato e amônio nas partes da planta forrageira e a distribuição percentual de nitrato e amônio entre parte aérea e raízes na Brachiaria brizantha cv. Marandu, submetida a combinações de doses de nitrogênio e potássio. O experimento foi realizado em casa de vegetação em solução nutritiva empregando-se sílica como substrato no período de dezembro de 2007 a fevereiro de 2008. Utilizou-se o esquema fatorial 52 fracionado, com 13 combinações de nitrogênio e potássio, as quais foram distribuídas segundo o delineamento de blocos ao acaso, com quatro repetições. Foram realizados dois cortes nas plantas. A interação doses de nitrogênio x doses de potássio foi significativa para número de perfilhos e de folhas, área foliar, massa seca da parte aérea e de raízes, comprimento e superfície total radicular e comprimento e superfície específica das raízes. A produção de massa seca da parte aérea teve forte correlação positiva com os parâmetros produtivos. As doses de nitrogênio se mostraram moduladoras do desenvolvimento do sistema radicular, o comprimento e superfície específicos diminuíram com emprego de altas doses de nitrogênio e potássio. A interação doses de nitrogênio x doses de potássio foi significativa para as concentrações de nitrogênio, potássio, nitrato e amônio em lâminas de folhas recém-expandidas e nas raízes. A atividade da enzima redutase do nitrato sofreu incremento com aumento das doses de nitrogênio e de potássio, em si. A distribuição percentual de nitrato e amônio ocorreu na ordem: colmos mais bainhas > lâminas de folhas recém-expandidas > folhas em expansão > lâminas de folhas maduras, evidenciando os colmos mais bainhas como parte da planta de acúmulo de nitrato e amônio e de mais baixa atividade da enzima redutase do nitrato. As doses de nitrogênio mostraram-se grande influenciadoras do valor SPAD, que aumentou com incremento de nitrogênio na solução nutritiva. O nitrogênio e o potássio alteram as características morfogênicas do capim-Marandu sendo determinantes para produção de massa seca da parte aérea e das raízes e que o desenvolvimento do sistema radicular é dependente de adequada nutrição, principalmente com nitrogênio. O nitrogênio e o potássio influenciaram o estado nutricional do capim-Marandu sendo determinantes para incremento do valor SPAD, atividade da enzima redutase do nitrato e concentração de nitrogênio, potássio, nitrato e amônio na parte aérea e nas raízes dessa gramínea. / Nitrogen and potassium are essentials for plant development, are extracted by plants in high amounts and their availabilities may change the morphogenetic, productive and nutritional characteristics of plants. The objectives of this research were: a) to assess fertilization with nitrogen and potassium rates on morphogenetic and shoot and root dry matter production of Brachiaria brizantha cv. Marandu; b) to evaluate the nitrogen, potassium, nitrate and ammonium concentration in the recently expanded leaf laminae and in the roots, SPAD values, and nitrate reductase activity, and ammonium and nitrate distribution in parts of Marandu palisadegrass in response to nitrogen and potassium rates combinations in nutrient solution. A greenhouse experiment was carried out, using ground quartz as a substrate, from December/2007 to February/2008. The experimental design used was fractionated 52 factorial, with 13 combinations of nitrogen and potassium rates, set in randomized block design with four replications. Two harvests were done in the plants. The nitrogen rates x potassium rates interaction was significant for the number of tiller, number of leaves, leaf area, above-ground, roots dry matter, total and specific root length, and total and specific root surface. Forage grass above-ground yield was positively correlated with number of tillers, number of leaves and leaf area. Nitrogen rates modulated the root system development, and the root specific length and surface decreased when high rates of both nitrogen and potassium were applied. Nitrogen rates x potassium rates interaction was significant for nitrogen, potassium, nitrate and ammonium concentrations in the recently expanded leaf laminae and in the roots. Nitrate reductase activity was increased with increasing the rates of both nitrogen and potassium. Nitrate and ammonium content in plant shoot were distributed following the order: culms+sheaths > recently expanded leaf laminae > emergent leaves > mature leaf laminae, showing culms+sheaths as a plant part of nitrate and ammonium a low nitrate reductase activity. Nitrogen rates highly influenced SPAD values, which increased with nitrogen rates in the nutrient solution. Nitrogen and potassium may affect the morphogenetic characteristics of Marandu palisadegrass, which were determinant for above-ground and root dry matter. The development of the root system depended on adequate plant nutrition, mainly in nitrogen. Nitrogen and potassium influenced the mineral nutrition status of Marandu palisadegrass and they increased the SPAD values, nitrate reductase activity, and nitrogen, potassium, nitrate and ammonium concentrations in shoots and roots of grass.

Made available in DSpace on 2017-07-10T17:37:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-09-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Citronella of Java (Cymbopogon winterianus Jowitt) is an aromatical plant with large usage in the manufacture of mosquito repellents because it contains essential oil rich in citronelal. The application of tissue culture on a large scale in medicinal plants production has increased significantly, since the conventional methods of propagation limit the potential use of some plants. Due that, one citronella of Java protocol was established with the objective of getting an alternative technique for the propagation of that culture. Four experiments ware conducted to evaluate the asepsis time, the explant type, the effect of vegetal regulators on the "in vitro" propagation of citronella of Java, as well as the micro-propagated seedling acclimatization. The gotten results had disclosed that the use of sodium hypochlorite (0.5%) during 30 minutes promoted a satisfactory asepsis index. The shoot tips explants type showed greater potential for the ?in vitro? development. It was verified that the use of benzilaminapurine (BAP) cytokynin is indispensable to succeed in the citronella of Java "in vitro" establishment, being able to be used in the concentration of 2.0 mg L-1. However, the use of indolbutyric acid (IBA) in the different tested concentrations did not present any results that justify its use, neither in the citronella of Java multiplication nor in the root establishment. The root establishment was evidenced when micro-propagated seedlings had been inoculated only in MS culture medium. It was verified that the check (without vegetal regulator) had a direct organogenesis, while such event was not verified in the other treatments. In the micro-propagated seedling acclimatization it was verified that the use of the floating method, it was essential for the seedlings survival, resulting in high survival index / A citronela de Java (Cymbopogon winterianus Jowitt) é uma planta aromática muito utilizada na fabricação de repelentes contra mosquitos por conter óleo essencial rico em citronelal. A utilização da cultura de tecidos, como forma de propagação de plantas medicinais tem aumentado significativamente, pois os métodos convencionais de propagação limitam o potencial de uso de algumas plantas. Sendo assim, um protocolo regenerativo para a citronela de Java foi estabelecido com o objetivo de se obter uma técnica alternativa para a propagação dessa cultura. Para isso foram realizados quatro experimentos envolvendo a avaliação do tempo de assepsia, do tipo de explante, do efeito de reguladores vegetais na propagação in vitro da citronela de Java, bem como a aclimatação das plântulas micropropagadas. Os resultados obtidos revelaram que a utilização do hipoclorito de sódio (0,5%) por 30 minutos promoveu índice satisfatório de assepsia. Os explantes com maior potencial ao desenvolvimento in vitro foram os ápices caulinares. Verificou-se que o emprego da citocinina benzilaminopurina (BAP) é indispensável para o sucesso do estabelecimento in vitro da citronela de Java, podendo ser utilizado na concentração de 2,0 mg L-1. No entanto, a utilização do ácido indolbutírico (IBA) nas diferentes concentrações testadas não apresentou resultados que justifique seu emprego, tanto na multiplicação quanto no enraizamento das plântulas de citronela de Java. O enraizamento foi evidenciado quando as plântulas micropropagadas foram inoculadas somente em meio de cultura MS. Foi constatado que na testemunha (sem regulador vegetal) houve uma organogênese direta, não sendo verificada nos demais tratamentos. Na aclimatação das plântulas micropropagadas verificou-se que a utilização do método floating , foi fundamental para a sobrevivência das plântulas, tendo sido observado alto índice de sobrevivência

The response of cucumber (Cucumis sativus L.) plants to various root and shoot environments (solution depth, temperature, ionic strength, nitrogen and calcium level and light intensity) were studied. Cucumber plants were grown in continuously circulating-solution in a heated-glasshouse. Dry weights of leaves, stems and roots, leaf area, leaf number, root length and root number were measured as well as uptake of potassium, calcium and nitrogen. The relationship between shoot and root in terms of functional equilibrium equations was also examined. The results presented show that: 1. Shoot growth of cucumber plants was reduced if grown in solutions of less than 50mm in depth; 2. When roots were grown in shallow solution depths at 1 or 5mm the dry weight allocated to the root increased. The ratio of root number/root length(no./cm) also increased. Lowering solution temperature to 12.5±2.5°C enhanced the production of root number relctive to root length, and 5 and 2% of full strength and 5% of full strength nitrogen level solution stimulated the growth of root length relative to root number; 3. Under low solution temperature treatment leaf number was maintained at the expense of leaf area. Under low total ionic strength and low nitrogen solution, enhanced root length growth was at the expense of leaf area growth; 4. Low solution temperature enhanced the dry weight allocated to the stem relative to the leaf. Low total ionic strength and low nitrogen solution increased the dry weight allocated to the leaf relative to the stem; 5. The specific activity of root, represented by specific absorption rate, increased when the shoot was under light stress and, the specific activity of shoot, represented by unit shoot rate, increased when the root was under nitrogen-stress; 6. The form of equation developed by Thornley (ΔM = fmΔW, where ΔM is the increment in weight of element M and ΔW the increment in total plant dry weight during a time period Δt with fm a constant) showed a better relationship than the equation developed by Davidson [root mass x rate(absorption) ∝ leaf mass x rate(photosynthesis)] and subsequently used by Hunt in the form of mass ratio(root/shoot) ∝ l/activity ratio; 7. The equation developed by Chung et al, total plant weight/(leaf number/leaf area) ∝ total "k"/(root number/root length), where k represents the total contents of elements or compounds, showed a good approximation of the relationship between shoot and root under all the environmental stresses imposed with the exception of calcium uptake. The results support the concept that the activity of the root or shoot in carrying out its function is influenced by the demand created by the opposite organ and appears to be a better assumption than that which proposes that the activity of an organ is solely dependent on its own size.

<p>About 70% of the plant species in the temperate zone are characterised by clonal growth, clonal species are also in majority in the Arctic and Subarctic where they affect the structure and composition of the vegetation. It is therefore of great importance to increase our knowledge about clonal plants and their growth and life histories. I have investigated how ramets of the stoloniferous plant <i>Linnaea borealis</i> are affected by the naturally occurring variation in environmental factors, such as: light, nutrient and water availability. Moreover, I examined the seed set and how supplemental hand pollination affects seed set in <i>L. borealis</i>, and also investigated the significance of the apical meristem for shoot population fitness. All studies were performed under field conditions in northern Sweden in a Subarctic environment and most are experimental.</p><p>The results show that nutrient resorption from senescing leaves is not significantly affecting the growth and nutrient pools of the ramet. This implies that the growth of<i> L. borealis </i>ramets is not governed by micro-site resource availability. However, removal of light competition resulted in increased branching and number of lateral meristems produced, reduced growth, and decreased root:shoot ratio on a per ramet basis. Thus, ramets of <i>L. borealis </i>can efficiently exploit favourable light patches through plastic growth. Apical dominance exerts a significant effect on shoot population fitness and can be lost through rodent grazing. However, loss of apical dominance is dependent on the timing of grazing, if the apical meristem is removed early in the autumn the ramet can repair the loss until the next summer. If grazing occur during spring the dry weight and leaf area production is affected negatively. Seed production in <i>L. borealis</i> in the Abisko area varies between years and sites, and was unaffected by supplemental hand pollination treatment, implying that there is no lack of pollinator activity.</p>

About 70% of the plant species in the temperate zone are characterised by clonal growth, clonal species are also in majority in the Arctic and Subarctic where they affect the structure and composition of the vegetation. It is therefore of great importance to increase our knowledge about clonal plants and their growth and life histories. I have investigated how ramets of the stoloniferous plant Linnaea borealis are affected by the naturally occurring variation in environmental factors, such as: light, nutrient and water availability. Moreover, I examined the seed set and how supplemental hand pollination affects seed set in L. borealis, and also investigated the significance of the apical meristem for shoot population fitness. All studies were performed under field conditions in northern Sweden in a Subarctic environment and most are experimental. The results show that nutrient resorption from senescing leaves is not significantly affecting the growth and nutrient pools of the ramet. This implies that the growth of L. borealis ramets is not governed by micro-site resource availability. However, removal of light competition resulted in increased branching and number of lateral meristems produced, reduced growth, and decreased root:shoot ratio on a per ramet basis. Thus, ramets of L. borealis can efficiently exploit favourable light patches through plastic growth. Apical dominance exerts a significant effect on shoot population fitness and can be lost through rodent grazing. However, loss of apical dominance is dependent on the timing of grazing, if the apical meristem is removed early in the autumn the ramet can repair the loss until the next summer. If grazing occur during spring the dry weight and leaf area production is affected negatively. Seed production in L. borealis in the Abisko area varies between years and sites, and was unaffected by supplemental hand pollination treatment, implying that there is no lack of pollinator activity.

In this thesis, optimization approaches for intensity-modulated radiation therapy are developed and evaluated with focus on numerical efficiency and treatment delivery aspects. The first two papers deal with strategies for solving fluence map optimization problems efficiently while avoiding solutions with jagged fluence profiles. The last two papers concern optimization of step-and-shoot parameters with emphasis on generating treatment plans that can be delivered efficiently and accurately. In the first paper, the problem dimension of a fluence map optimization problem is reduced through a spectral decomposition of the Hessian of the objective function. The weights of the eigenvectors corresponding to the p largest eigenvalues are introduced as optimization variables, and the impact on the solution of varying p is studied. Including only a few eigenvector weights results in faster initial decrease of the objective value, but with an inferior solution, compared to optimization of the bixel weights. An approach combining eigenvector weights and bixel weights produces improved solutions, but at the expense of the pre-computational time for the spectral decomposition. So-called iterative regularization is performed on fluence map optimization problems in the second paper. The idea is to find regular solutions by utilizing an optimization method that is able to find near-optimal solutions with non-jagged fluence profiles in few iterations. The suitability of a quasi-Newton sequential quadratic programming method is demonstrated by comparing the treatment quality of deliverable step-and-shoot plans, generated through leaf sequencing with a fixed number of segments, for different number of bixel-weight iterations. A conclusion is that over-optimization of the fluence map optimization problem prior to leaf sequencing should be avoided. An approach for dynamically generating multileaf collimator segments using a column generation approach combined with optimization of segment shapes and weights is presented in the third paper. Numerical results demonstrate that the adjustment of leaf positions improves the plan quality and that satisfactory treatment plans are found with few segments. The method provides a tool for exploring the trade-off between plan quality and treatment complexity by generating a sequence of deliverable plans of increasing quality. The final paper is devoted to understanding the ability of the column generation approach in the third paper to find near-optimal solutions with very few columns compared to the problem dimension. The impact of different restrictions on the generated columns is studied, both in terms of numerical behaviour and convergence properties. A bound on the two-norm of the columns results in the conjugate-gradient method. Numerical results indicate that the appealing properties of the conjugate-gradient method on ill-conditioned problems are inherited in the column generation approach of the third paper. / QC 20100709

Endophytes are organisms that live inside plants without causing disease and include microbes that benefit their hosts by aiding in nutrient acquisition and pathogen control. This thesis concerns the endophytes of the genus Zea which includes modern maize (Zea mays L.). Beginning 9,000 years ago, maize was domesticated from wild grasses in Mexico (teosintes), bred into diverse varieties and moved to new soils throughout the Americas. The impact of these long-term changes on the associated endophytic communities has not been examined. Furthermore, today, maize is routinely transplanted around the world to facilitate breeding, but the short-term impact of switching soils on endophyte composition is not known. I attempted to answer the first question by surveying the bacterial endophytes that inhabit 14 diverse ancestral, ancient and modern Zea genotypes. To answer the second question, three extreme Zea genotypes, ancestral, intermediate and modern, were grown side by side on two extreme soils that span the tropical-to-temperate migration route of maize. Endophyte populations from seeds, roots and shoots were DNA fingerprinted using terminal restriction length polymorphism (TRFLP) of 16S rDNA. To understand microbial functions, bacteria were cultured and tested for >13 in vitro traits including nitrogen fixation, phosphate solubilization, plant hormone production and antibiosis. Relationships between endophyte communities were analyzed using principle component analysis (PCA) and Sᴓrensen’s similarity index. The results show that different Zea tissues and genotypes have diverse endophytic communities. The community composition of seed endophytes correlates with host phylogeny suggesting that as humans bred maize, they inadvertently impacted its microbial inhabitants, though the change was gradual. Soil swapping and growth on sterile sand confirm that shoot and root endophyte communities in juvenile plants are primarily inherited. However, a given maize genotype can also select and take up the same microbes (based on TRFLP) from geographically diverse soils. These latter results show that the endophyte communities of Zea plants are significantly buffered from the short-term effects of migration. A few microbes and microbial traits are conserved across all Zea genotypes and soil treatments, suggestive of a core taxonomic and functional microbiota for this agriculturally important genus. / OMAFRA New Directions, Ontario Research Fund, Canadian Foundation for Innovation

The ratio of auxin to cytokinin determines de novo organogenesis in plants. Relatively little is known about the effect of genetically altered auxin signaling on in vitro organogenesis. Here, callusogenesis, shoot, and root formation were studied in loss- (LOF) and gain-of-function (GOF) alleles in two phylogenetically related Auxin Response Factors (ARFs), MONOPTEROS (MP/ARF5) and NON-PHOTOTROPHIC HYPOCOTYL 4 (NPH4/ARF7). Reduced MP activity greatly diminished shoot regeneration, and partially diminished callusogenesis and root formation. LOF in NPH4 strongly decreased callusogenesis, and mildly decreased shoot and root regeneration in particular categories of explants. By contrast, organogenesis responses were strongly increased in aerial explants carrying the GOF transgene dMP. Thus, both MP and NPH4 seem to act as positive regulators of certain organogenesis processes and the GOF dMP transgene may be of interest for stimulating organogenesis in plant species with poor regeneration properties. Also, organogenesis in vitro may reveal unknown developmental ARF functions.

The ratio of auxin to cytokinin determines de novo organogenesis in plants. Relatively little is known about the effect of genetically altered auxin signaling on in vitro organogenesis. Here, callusogenesis, shoot, and root formation were studied in loss- (LOF) and gain-of-function (GOF) alleles in two phylogenetically related Auxin Response Factors (ARFs), MONOPTEROS (MP/ARF5) and NON-PHOTOTROPHIC HYPOCOTYL 4 (NPH4/ARF7). Reduced MP activity greatly diminished shoot regeneration, and partially diminished callusogenesis and root formation. LOF in NPH4 strongly decreased callusogenesis, and mildly decreased shoot and root regeneration in particular categories of explants. By contrast, organogenesis responses were strongly increased in aerial explants carrying the GOF transgene dMP. Thus, both MP and NPH4 seem to act as positive regulators of certain organogenesis processes and the GOF dMP transgene may be of interest for stimulating organogenesis in plant species with poor regeneration properties. Also, organogenesis in vitro may reveal unknown developmental ARF functions.