Thesis (Ph. D. (Botany))--Rhodes University, 2006.

Our planet is growing rapidly in population and with that comes a demand for resources. To address issues in food security, scientists are looking to the underground parts of plants for novel mechanisms that will eventually lead to enhanced crop traits. Scientists are examining the underlying genetic frameworks to identify which genes play key roles in specific developmental processes. In this study we examined the model plant Arabidopsis thaliana as the roots are easily visualised, the genome has been sequenced, and there are many tools broadly available to work with. This thesis has used a multidisciplinary approach to uncover the signalling cascades revolving around the small signalling peptide RALF34, which is significantly involved in primary and lateral root growth. We have demonstrated, in the following chapters that RALF peptides are inherent to normative lateral root initiation, potentially regulated through shoot derived auxin. Furthermore, RALF4 and 34 peptides play a strong role in restricting primary root growth, and that together these peptides have an additive effect on cell elongation. Lastly, we identify several leucine-rich repeat receptor-like proteins, kinase proteins, and cell wall remodelling enzymes, which putatively play unique and diverse roles during primary and lateral root development. Taken together, this thesis provides novel and unique insights into new signalling pathways during root growth, which may in future aid in agronomic enterprises.

In flowering plants, the opening of the anther to release pollen is carefully timed to maximise reproductive potential. Manipulation of this process is an important tool for plant breeding and the production of hybrid crops. Dehydration of the anther epidermis, combined with the presence of secondary thickening within the endothecium layer, is required to create biomechanical changes that enable anther dehiscence. Both passive and active processes contribute to the targeted removal of water from the anther walls, however the genetic factors controlling water movement are not known. Furthermore, the presence of stomata on anthers may enhance water loss via evaporation. In plants, active movement of water can be achieved by regulation of water channels and by changes to the osmotic potential of organs; this was explored in the context of changes in the anther driving anther dehiscence and pollen release. qRT-PCR analysis was used to identify aquaporin and sucrose transporter genes that are upregulated during anther dehiscence in Arabidopsis thaliana. For genes of interest, the phenotypes of available mutants were characterised. Combinations of single, double and triple mutants showed changes in fertility and variations in floral organ lengths. Analysis of GUS reporter lines showed that the promoter activity of different aquaporins is confined to specific parts of the flower. The results suggest that certain aquaporins isoforms enhance hydraulic conductivity in different parts of the flower, which could contribute to water transport required for petal and filament extension. The importance of evaporation during anther dehydration was also investigated. The phenotypes of Arabidopsis mutant lines with varying stomatal densities were characterised, and changes in fertility were investigated under different environmental conditions. High relative humidity delayed anther dehiscence and affected pollen viability, resulting in reduced fertility. Plants that have no anther stomata were most severely affected. These results suggest that water loss via evaporation is important for anther opening in Arabidopsis, and that the presence of stomata on anthers facilitates this process.

Made available in DSpace on 2014-06-11T19:27:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-04-27Bitstream added on 2014-06-13T19:35:33Z : No. of bitstreams: 1 lara_not_me_botib_parcial.pdf: 279145 bytes, checksum: 65872f7d6809e41d2423f028c211d092 (MD5) Bitstreams deleted on 2015-06-03T11:42:34Z: lara_not_me_botib_parcial.pdf,. Added 1 bitstream(s) on 2015-06-03T11:44:02Z : No. of bitstreams: 1 000706739_20150701.pdf: 265984 bytes, checksum: 671c09d297b1107bbf2470829f6b7fdc (MD5) Bitstreams deleted on 2015-07-01T11:47:26Z: 000706739_20150701.pdf,. Added 1 bitstream(s) on 2015-07-01T11:48:09Z : No. of bitstreams: 1 000706739.pdf: 1918406 bytes, checksum: 80e942f41721ae04d65f0e8edcc1bec2 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A família Rubiaceae é a quarta maior família entre as eudicotiledôneas, possui distribuição pantropical com representantes arbustivos a arbóreos, sendo as regiões sul-americanas aquelas que superam todas as regiões do mundo em diversidade de espécies da família. Em ambientes tão diversos, um conjunto de fatores pode influenciar, direta ou indiretamente, a atividade do câmbio vascular nas espécies arbóreas desses ambientes e consequentemente em seus produtos, o xilema e floema secundários. Assim, este estudo abrange a caracterização do sistema vascular secundário bem como a atividade cambial e a diferenciação celular em relação à fenologia da espécie e às condições climáticas da região. Para tanto, foi escolhida a espécie Alibertia concolor (Cham) K. Schum. (Rubiaceae) por apresentar camadas de crescimento bem demarcadas no lenho, com potencial para análise de atividade cambial e periodicidade de crescimento. Foram realizadas seis coletas em cinco espécimes em área de floresta estacional semidecidual no Jardim Botânico da UNESP, Campus de Botucatu, no ano de 2010. Xilema e floema secundários mostram um paralelismo entre suas características anatômicas. O xilema secundário da espécie apresenta camadas de crescimento demarcadas por faixas marginais de parênquima axial, enquanto que o floema secundário apresentou faixas tangenciais de fibras intercalando as camadas de células vivas do floema. Porosidade difusa foi visualizada nos vasos do xilema secundário e também não foi observada diferença no tamanho dos elementos de tubo crivado no floema secundário quando comparados os floemas inicial e tardio. Os elementos de vaso do xilema secundário possuem placa de perfuração simples e os elementos de tubo crivado, por sua vez, possuem placas crivadas simples. Fibras gelatinosas... / The family Rubiaceae is the fourth largest family between eudicotyledonous. It has a pantropical distribution with shrubs to trees. In South American regions the family surpass all regions of the world in species diversity. In such diverse environments a set of factors can influence, directly or indirectly, the activity of vascular cambium in woody species of this family, and thus their products, the secondary xylem and phloem. The goal of this study was to characterize the secondary vascular system in one of the representative species from Rubiaceae, Alibertia concolor (Cham) K. Schum., its cambial activity and cell differentiation in relation to the phenology of the species and climatic conditions of the region. The species was selected because the growth rings in the wood were well-defined which suggests growth periodicity. We collected periodically samples of five specimens in a semideciduous forest at the Botanical Garden of UNESP, Botucatu, in the year 2010. Secondary xylem and phloem show a parallel between their anatomical characteristics. Marginal bands of axial parenchyma delimits growth rings in the xylem while tangential fiber bands are present in the phloem. Diffuse porosity in the xylem and no difference in the size of sieve tube elements in the phloem. Simple perforation plates in vessel elements, and simple sieve plates in sieve tube elements. Rays are predominantly uniseriate and biseriate in the xylem as well as in the phloem. The species is evergreen, however, there is a remarkable period of growth and sprouting of buds in December. Flower buds and flowering were observed in January-February. The initiation of cambial activity occurred in December, and it was related to the first rains and budding. The ending of cambial activity occurred in February, rainy season, and cambium remained dormant until October. In the... (Complete abstract click electronic access below)

Orientador: Carmen Regina Marcatti / Banca: Patricia Soffiatti / Banca: Tatiane Maria Rodrigues / Resumo: A família Rubiaceae é a quarta maior família entre as eudicotiledôneas, possui distribuição pantropical com representantes arbustivos a arbóreos, sendo as regiões sul-americanas aquelas que superam todas as regiões do mundo em diversidade de espécies da família. Em ambientes tão diversos, um conjunto de fatores pode influenciar, direta ou indiretamente, a atividade do câmbio vascular nas espécies arbóreas desses ambientes e consequentemente em seus produtos, o xilema e floema secundários. Assim, este estudo abrange a caracterização do sistema vascular secundário bem como a atividade cambial e a diferenciação celular em relação à fenologia da espécie e às condições climáticas da região. Para tanto, foi escolhida a espécie Alibertia concolor (Cham) K. Schum. (Rubiaceae) por apresentar camadas de crescimento bem demarcadas no lenho, com potencial para análise de atividade cambial e periodicidade de crescimento. Foram realizadas seis coletas em cinco espécimes em área de floresta estacional semidecidual no Jardim Botânico da UNESP, Campus de Botucatu, no ano de 2010. Xilema e floema secundários mostram um paralelismo entre suas características anatômicas. O xilema secundário da espécie apresenta camadas de crescimento demarcadas por faixas marginais de parênquima axial, enquanto que o floema secundário apresentou faixas tangenciais de fibras intercalando as camadas de células vivas do floema. Porosidade difusa foi visualizada nos vasos do xilema secundário e também não foi observada diferença no tamanho dos elementos de tubo crivado no floema secundário quando comparados os floemas inicial e tardio. Os elementos de vaso do xilema secundário possuem placa de perfuração simples e os elementos de tubo crivado, por sua vez, possuem placas crivadas simples. Fibras gelatinosas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The family Rubiaceae is the fourth largest family between eudicotyledonous. It has a pantropical distribution with shrubs to trees. In South American regions the family surpass all regions of the world in species diversity. In such diverse environments a set of factors can influence, directly or indirectly, the activity of vascular cambium in woody species of this family, and thus their products, the secondary xylem and phloem. The goal of this study was to characterize the secondary vascular system in one of the representative species from Rubiaceae, Alibertia concolor (Cham) K. Schum., its cambial activity and cell differentiation in relation to the phenology of the species and climatic conditions of the region. The species was selected because the growth rings in the wood were well-defined which suggests growth periodicity. We collected periodically samples of five specimens in a semideciduous forest at the Botanical Garden of UNESP, Botucatu, in the year 2010. Secondary xylem and phloem show a parallel between their anatomical characteristics. Marginal bands of axial parenchyma delimits growth rings in the xylem while tangential fiber bands are present in the phloem. Diffuse porosity in the xylem and no difference in the size of sieve tube elements in the phloem. Simple perforation plates in vessel elements, and simple sieve plates in sieve tube elements. Rays are predominantly uniseriate and biseriate in the xylem as well as in the phloem. The species is evergreen, however, there is a remarkable period of growth and sprouting of buds in December. Flower buds and flowering were observed in January-February. The initiation of cambial activity occurred in December, and it was related to the first rains and budding. The ending of cambial activity occurred in February, rainy season, and cambium remained dormant until October. In the... (Complete abstract click electronic access below) / Mestre

An investigation of the anatomy and gas exchange characteristics of Mariscus congestus in three different habitats was undertaken in order to establish whether M. congestus from the three different habitats displayed any ecotypic responses when placed in a new similar environment. It was hoped that the results of this investigation would yield evidence that would support the ecotype concept similar to the investigations of Milner and Hiesey (1964), Green (1969) and Slayter and Ferrar (1977). On the basis of the site leaf anatomy, M. congestus investigated at the coast (site 1) differed in many respects from the inland plants (sites 2 and 3). These differences suggest that the coastal plants may have undergone a slight ecotypic divergence from the inland plants. The anatomical investigation also suggested that the leaves of M. congestus from all three sites may either be C₄ NADP-ME or PCK and that all had typical Chlorocyperiod anatomy. The habitat microclimates at sites 1-3 had different light and water regimes. There were no significant differences between the 12 month temperature environments of the three sites. There was however, a minor difference between the coastal (high temperature) and the inland (lower temperature) sites. M. congestus at the three sites had significantly different CO₂ assimilation rates, transpiration and stomatal conductance in response to the differing habitat microclimates. The water use efficiency of the sites were however, similar. Site 1 attained the highest CO₂ assimilation rates, transpiration, stomatal conductance, and water use efficiency and site 3 the lowest. Under similar conditions the gas exchange data for the potted plants indicated that M. congestus from the different sites was typically C₄. The optimal photosynthetic temperatures of all the sites was above 30°C and they did not show significant inhibition of CO₂ assimilation by different oxygen concentrations. The results of the laboratory investigation of the potted plants suggested that the only site-specific (ecotypic) response of M. eongestus was the light intensity at which the plants from the different sites were light saturated. The light and temperature response of field plants under field conditions was not comparable to the light and temperature response of potted plants under laboratory conditions. This may have been due to the field results being obtained under differing water and soil nutrient regimes. The potted plants may also have had a reduced root mass compared to their field counterparts and the potted plants may have also have become root bound. Under field conditions the plants had differing light saturation points and optimal photosynthetic temperatures compared to the potted plants. This investigation thus did not support the hypothesis stated in this thesis. The data in this investigation thus may indicate that plants with as diverse habitats as Mariscus congestus that are removed from their natural habitats display rapid changes in gas exchange characteristics in response to their new microclimates, with few ecotypic physiological characteristics of the old habitat being retained.

The roots of a plant play a vital role in its growth and development, but due to practical difficulties of observing underground roots, the study of their shape has long been neglected. Recent advances in CT imaging technology have allowed for accurate non-destructive imaging of root systems in soil. This technique has formed the basis of the FutureRoots project. The main challenge with analysing the shape of a plant root system is that they have varying topological structure, so traditional shape analysis methods cannot be applied. In this thesis, we develop three approaches for analysing wheat root systems. The first approach involves measuring a set of pre-chosen root traits, and analysing this set using conventional statistical methods. This approach is effective but may miss potentially important shape information and the large number of measurable traits reduces the potential power of statistical tests. The second approach is to perform pairwise comparisons based on the Hausdorff Metric and use Multidimensional scaling to reduce a large set of pairwise comparisons to a dataset which can be analysed with conventional statistical methods. This approach can detect and test for overall shape differences but can fail to detect subtle differences. The third approach is to apply the Persistent Homology technique from Topological Data Analysis, which is designed to find underlying topological differences between two shapes. This method successfully finds differences but it is difficult to interpret the results. We will apply these three techniques to simulated data and a real life dataset. In addition, because of experimental considerations, the wheat roots had to be unnaturally constrained to a small area so we have developed a method to estimate how they would have grown unconstrained.

X-ray micro computed tomography (µCT) is increasingly applied in plant biology as an imaging system that is valuable for the study of root development in soil, since it allows the three-dimensional and non-destructive visualisation of plant root systems. Variations in the X-ray attenuation values of root material and the overlap in measured intensity values between roots and soil caused by water and organic matter represent major challenges to the extraction of root system architecture. We propose a novel technique to recover root system information from X-ray CT data, using a strategy based on a visual tracking framework embedding a modiffed level set method that is evolved using the Jensen-Shannon divergence. The model-guided search arising from the visual tracking approach makes the method less sensitive to the natural ambiguity of X-ray attenuation values in the image data and thus allows a better extraction of the root system. The method is extended by mechanisms that account for plagiatropic response in roots as well as collision between root objects originating from different plants that are grown and interact within the same soil environment. Experimental results on monocot and dicot plants, grown in different soil textural types, show the ability of successfully extracting root system information. Various global root system traits are measured from the extracted data and compared to results obtained with alternative methods.

Classical Morphology in Plant Sciences requires a typological view of plant organs. This usually implies in classifying stem, leaf and root as basic and well-defined unities. Ferns are the most diverse group of non-flowering plants and occupy a key position in the land plants phylogeny. Their leaves are usually understood as homologous to those of seed-plants. Still, they bear intriguing features, like a leaf apical meristem bearing a distinct apical cell, and that may be many times divided, resembling the activity of a whole shoot. We present a study about the leaf development in some leptosporangiate ferns of different morphologies to better understand how these structures may have evolved and the possible homologies between their ontogenetic processes. Class I KNOX genes expression was analyzed in the heteroblastic fern Mickelia scandens, as they are related to organ determinacy in angiosperms. The two copies of Class I KNOX are expressed even in determined structures, like pinnae. But a reduction of the quantity of transcript is related to the development of the less determinate frond form that occurs in terrestrial individuals. Using classic anatomical tools, we studied the development of leaves in ferns related to Mickelia scandens that present different morphologies. In addition, we observed natural occurring mutants in a collection. The basic structure of apical cells is essentially well conserved in all the group. Marginal cells, classically pointed as part of the marginal meristem, may repeat in some degree the activity of the leaf apical cell. Changes in the structure and activity of these structures may be the reason why simple-leaved ferns of the genus Elaphoglossum do not make compound leaves and why usual leaf morphology may change, producing anomalous structures. We discuss this data based on Agnes Arber concepts of partial-shoot and identity-in-parallel, proposing an interpretation of the fern leaf not as a well-defined organ, but a product of ontogenetic processes, some of them typical of the shoot / A Morfologia Clássica em Botânica requer uma visão tipológica dos órgãos vegetais. Isso geralmente implica na classificação de caule, folha e raiz como unidades básicas e bem definidas. Samambaias são o grupo mais diverso de plantas sem flores e ocupam uma posição-chave na filogenia das plantas terrestres. Suas folhas geralmente são entendidas como homólogas às de espermatófitas. Ainda assim, possuem características intrigantes, como um meristema apical foliar com uma célula apical distinta, e podem ser muitas vezes divididas, lembrando a atividade de um sistema caulinar. Apresentamos um estudo do desenvolvimento foliar em algumas samambaias leptosporangiadas de diferentes morfologias para entender melhor como essas estruturas podem ter evoluído e as possíveis homologias entre seus processos ontogênicos. A expressão dos genes de Classe I KNOX foi analisada na samambaia heteroblástica Mickelia scandens, uma vez que estão relacionados à determinação de órgãos em angiospermas. As duas cópias de Classe I KNOX são expressas mesmo em estruturas determinadas, como pinas. Mas uma redução da quantidade de transcritos está relacionada ao desenvolvimento da forma menos determinada da fronde que ocorre em indivíduos terrestres. Usando ferramentas anatômicas clássicas, estudamos o desenvolvimento de folhas em samambaias relacionadas a Mickelia scandens que apresentam diferentes morfologias. Além disso, observamos mutantes de ocorrência natural em uma coleção. A estrutura básica das células apicais é essencialmente bem conservada em todo o grupo. Células marginais, classicamente apontadas como parte do meristema marginal, podem repetir em certo grau a atividade da célula apical da folha. Mudanças na estrutura e atividade dessas estruturas podem ser a razão pela qual a samambaia de folhas simples do gênero Elaphoglossum não fazem folhas compostas e porque a morfologia de uma folha normal pode ser alterada, produzindo estruturas anômalas. Discutimos esses dados com base em conceitos de Agnes Arber de sistema caulinar-parcial identidade-em-paralelo, propondo uma interpretação da folha de samambaia não como um órgão bem definido, mas como um produto de processos de ontogênese, alguns deles típicos do sistema caulinar

Orientador: Aline Redondo Martins / Banca: Mario Tomazello Filho / Banca: Mario Luiz Teixeira de Moraes / Resumo: Anacardiaceae possui aproximadamente 81 gêneros e 800 espécies, presentes em uma grande variedade de ambientes e climas. Astronium fraxinifolium é uma espécie arbórea, dióica encontrada facilmente no cerrado e que produz madeira de excelente qualidade, utilizada na construção civil. As espécies vegetais expressam características em resposta a pressão do meio em que se encontram. Espécies do cerrado podem apresentar características de ambientes xéricos, por exemplo. O presente trabalho teve como objetivo investigar se a densidade, os constituintes químicos estruturais e características anatômicas da madeira de A. fraxinifolium são influenciados pelo ambiente, uma vez que foram comparados os dados dos materiais coletados em uma área de solo degradado com os dados da mesma espécie em ambiente não degradado. As amostras de lenho foram submetidas a técnicas usuais de anatomia da madeira, bem como para determinação de sua densidade aparente e também quantificação dos constituintes químicos. As comparações se deram entre as áreas (impactada e não impactada) e entre indivíduos masculinos e femininos. Os resultados mostraram que as plantas crescidas na área impactada tiveram diferenças significativas na densidade aparente do lenho e em características anatômicas. Para a análise dos constituintes, a variação se deu principalmente em relação a posição de cerne-alburno. A espécie mostrou diferenças significativas entre indivíduos masculinos e femininos, em relação a anatomia e a densidad... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Anacardiaceae has approximately 81 genus and 800 species and it is distributed in a wide variety of environments and climates. Astronium fraxinifolium is a tree, dioecious, present in the cerrado and that produces excellent quality wood, used in the civil construction. Plant species express characteristics in response to the environmental pressure that allows their survive. Species of cerrado can present characteristics of xeric environments, for example. The objective of this work was to investigate whether the density, structural chemical constituents and anatomical characteristics of A. fraxinifolium wood are influenced by the environment, since the data of the collected materials were compared in a degraded soil area with the same species data in a non-degraded environment. The wood samples were submitted to the usual techniques of wood anatomy, as well as to determine their apparent density and quantification of the chemical constituents. The comparisons were between the areas (impacted and not impacted) and between male and female plants. The results showed that the plants grown in the impacted area had significant differences in the apparent density of the wood and in anatomical characteristics. For the analysis of the constituents, the variation was mainly in relation to the heartwood/sapwood position. The species showed significant differences between male and female individuals, relative to anatomy and density, but not to chemical constituents. With this data, we co... (Complete abstract click electronic access below) / Mestre