Thesis (Ph.D.)--University of Western Australia, 2006.

Thesis (Ph.D.)--University of Wollongong, 2006. / Typescript. Includes bibliographical references: p. 262-287.

Thesis (Ph.D.) - University of Queensland, 2005. / Includes bibliography.

[Truncated abstract] Grevillea and Leucadendron belong to Proteaceae and both have economic importance to the floriculture industry. Grevillea is a highly diverse genus endemic to Australia and very attractive for landscaping. Leucadendron is a South African Proteaceae but is cultivated in Australia and is well known as a cut flower. This thesis focuses on the application of DNA-based molecular markers to these genera. Several groupings within Grevillea were suggested by previous researchers based on morphological characteristics. In this thesis the monophyly of the groupings among 12 Grevillea species from New South Wales was tested using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses. To test the robustness of the data, UPGMA using Jaccard similarity, Neighbor Joining using total character difference and Wagner parsimony analyses were undertaken. The relationship trees generated supported monophyly of the groupings. Chloroplast DNA (cpDNA) was used to develop phylogenetic relationships among Leucadendron species. Inheritance and variation of cpDNA were evaluated using PCR-RFLP. The study demonstrated that cpDNA was inherited maternally and a phylogenetic tree of Leucadendron species using parsimony analysis was constructed. ... A fingerprinting study conducted using ISSR, produced a dendrogram showing the relationships among 30 cultivars. From the results, i a fingerprinting key was developed. Three examples of synonymous cultivar pairs were identified. In Leucadendron the male and female flowers develop on separate plants, and sex identification is only possible at time of flowering. ISSR, suppression subtractive hybridisation (SSH), and SSH combined with mirror orientation selection (MOS) were used in attempts of identifying sex-dependent DNA fragments at earlier stages of plant development. Neither of these techniques was able to identify sex-specific markers in Leucadendron. Nevertheless, the results did indicate that cpDNA copy number may differentiate male and female plants. Also, it was demonstrated that the genomes of male and female plants are quite homologous, which increases the difficulty in identifying sex-specific sequences. This thesis highlights the potential of DNA-based markers to determine species relationships in Grevillea and Leucadendron, as well as to identify Leucadendron cultivars. The information produced during the research for this thesis provides a basis for Grevillea and Leucadendron variety development and may be used to assist the design of interspecific crosses, to identify cultivars and the parents of hybrids. In addition, the results offer insights into the likelihood, problems and strategies of finding sex-specific markers for genes controlling sex in Leucadendron. ii

Thèse (M.Sc.)--Université Laval, 1998. / Bibliogr.: f. 59-64. Publié aussi en version électronique.

Thesis (Ph.D.)--University of Wollongong, 2004. / Typescript. Includes bibliographical references: p. 241-260.

Resumo: O presente trabalho teve por objetivo avaliar o emprego da reflectometria no domínio do tempo, conhecida também, como TDR (Time Domain Reflectometry) na obtenção da umidade em madeiras. Atualmente esta técnica é muito empregada na física dos solos e sistemas de irrigação, para o monitoramento do teor de água no solo. Para este estudo, empregou-se duas espécies de madeiras comerciais, o Pinus caribaea var. hondurensis e a Grevillea robusta (Cunn.), procedentes dos municípios de Cabrália Paulista-SP e Duartina-SP. Para cada espécie, foram escolhidas de forma aleatória 11 árvores onde estas foram abatidas, serradas em toras de 3,0m e encaminhadas para serraria onde foi realizado o processo de desdobramento em tábuas pelo sistema de corte tangencial. As tábuas apresentaram espessura de 28mm e largura de 24cm. Em seguida, escolheu-se novamente de forma aleatória 11 tábuas, que foram embaladas em plástico para evitar a perda de água, e encaminhadas para o laboratório. Uma vez obtidas as amostras representativas para avaliação, estas foram inicialmente pesadas e secas em estufa elétrica a 40 oC de temperatura. Para o Pinus caribaea var. hondurensis, o processo de secagem iniciou-se com umidade média de 101,45% e finalizou, à 7,36% de umidade. Já para a Grevillea robusta (Cunn.), a umidade média inicial foi de 125,88% e final de 6,62%. Durante o processo de secagem, a umidade foi determinada, periodicamente, através do método de massas correntes, bem como, a respectiva constante dielétrica relativa pela TDR. Para o estudo da viabilização da TDR na determinação... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The main objective of this present work was to evaluate the applied of the reflectometry in the domain of time, technique known as TDR (Time Domain Reflectometry) to obtention of moisture in woods. Nowadays, this technique is very used in soil physics and irrigation system, to the monitoring of moisture in the soil. To this study, two species of commercial woods were used, the Pinus caribaea var. hondurensis and the Grevillea robusta (Cunn.), these woods come from the municipal district of Cabrália Paulista-SP and Duartina-SP. To each specie, were chosen in a casual way, 11 trees. They were dejected, planked in logs of 3,0m and referred to the sawmill, the place where was realized the process of unfolding the wood in boards by the system of tangential section. The boards brung up the ply of 28mm and the width of 24cm. Next, were chosen again, in a casual way, 11 planks that were unrolled in plastic just to avoid the waterless, and referred to the lab. When were obtained the appropriate samples to the evaluation, they were, in an initial instant, measure out and dry in an electric greenhouse of a temperature of 40 oC. To Pinus caribaea var. hondurensis, the dry process started with a media moisture of 101,45% and finished with 7,36% of moisture. To the other specie, the Grevillea robusta (Cunn.), the initial media moisture found was 125,88% and the final was 6,62%. During the whole dry process, the moisture was determinated, periodically, throw the masses current, as like the relative dielectric constant by the TDR. To the study of the feasibility of TDR in the determination of moisture in woods, was chosen to establish 3 distances among the sensor's stick: 10mm, 20mm and 30mm (these distances were applied in commercial resistive gauges too). In the same way, it was established the variations in the velocity of propagation of electromagnetic pulses send out from TDR equipment... (Complete abstract click electronic access below) / Orientador: João Eduardo Guarnetti dos Santos / Coorientador: Antonio Evaldo Klar / Coorientador: Elias Taylor Durgante Severo / Banca: Marcos Antonio de Rezende / Banca: Momotaro Imaizumi / Banca: José Fernandes Presenço / Banca: Eik Teório / Doutor

Made available in DSpace on 2014-06-11T19:31:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-19Bitstream added on 2014-06-13T19:20:48Z : No. of bitstreams: 1 batista_wr_dr_botfca.pdf: 2260401 bytes, checksum: b61aab17efe4e099be5e84d8637f318a (MD5) / Outros / O presente trabalho teve por objetivo avaliar o emprego da reflectometria no domínio do tempo, conhecida também, como TDR (Time Domain Reflectometry) na obtenção da umidade em madeiras. Atualmente esta técnica é muito empregada na física dos solos e sistemas de irrigação, para o monitoramento do teor de água no solo. Para este estudo, empregou-se duas espécies de madeiras comerciais, o Pinus caribaea var. hondurensis e a Grevillea robusta (Cunn.), procedentes dos municípios de Cabrália Paulista-SP e Duartina-SP. Para cada espécie, foram escolhidas de forma aleatória 11 árvores onde estas foram abatidas, serradas em toras de 3,0m e encaminhadas para serraria onde foi realizado o processo de desdobramento em tábuas pelo sistema de corte tangencial. As tábuas apresentaram espessura de 28mm e largura de 24cm. Em seguida, escolheu-se novamente de forma aleatória 11 tábuas, que foram embaladas em plástico para evitar a perda de água, e encaminhadas para o laboratório. Uma vez obtidas as amostras representativas para avaliação, estas foram inicialmente pesadas e secas em estufa elétrica a 40 oC de temperatura. Para o Pinus caribaea var. hondurensis, o processo de secagem iniciou-se com umidade média de 101,45% e finalizou, à 7,36% de umidade. Já para a Grevillea robusta (Cunn.), a umidade média inicial foi de 125,88% e final de 6,62%. Durante o processo de secagem, a umidade foi determinada, periodicamente, através do método de massas correntes, bem como, a respectiva constante dielétrica relativa pela TDR. Para o estudo da viabilização da TDR na determinação... / The main objective of this present work was to evaluate the applied of the reflectometry in the domain of time, technique known as TDR (Time Domain Reflectometry) to obtention of moisture in woods. Nowadays, this technique is very used in soil physics and irrigation system, to the monitoring of moisture in the soil. To this study, two species of commercial woods were used, the Pinus caribaea var. hondurensis and the Grevillea robusta (Cunn.), these woods come from the municipal district of Cabrália Paulista-SP and Duartina-SP. To each specie, were chosen in a casual way, 11 trees. They were dejected, planked in logs of 3,0m and referred to the sawmill, the place where was realized the process of unfolding the wood in boards by the system of tangential section. The boards brung up the ply of 28mm and the width of 24cm. Next, were chosen again, in a casual way, 11 planks that were unrolled in plastic just to avoid the waterless, and referred to the lab. When were obtained the appropriate samples to the evaluation, they were, in an initial instant, measure out and dry in an electric greenhouse of a temperature of 40 oC. To Pinus caribaea var. hondurensis, the dry process started with a media moisture of 101,45% and finished with 7,36% of moisture. To the other specie, the Grevillea robusta (Cunn.), the initial media moisture found was 125,88% and the final was 6,62%. During the whole dry process, the moisture was determinated, periodically, throw the masses current, as like the relative dielectric constant by the TDR. To the study of the feasibility of TDR in the determination of moisture in woods, was chosen to establish 3 distances among the sensor’s stick: 10mm, 20mm and 30mm (these distances were applied in commercial resistive gauges too). In the same way, it was established the variations in the velocity of propagation of electromagnetic pulses send out from TDR equipment... (Complete abstract click electronic access below)

Proteins that recognise the sugar surface structures on cells have an enormous potential to be used as tools in the characterisation of these structures. A group of proteins, called lectins, have been identified that can bind to carbohydrate complexes on the receptors of cells. The crude extract from Grevillea robusta seeds was found to contain lectin-like proteins that were different from most other lectins, as they would specifically target the receptors of white blood cells and not those found on red blood cells. Therefore, the lectin isolated from G.robusta could be used as a tool to identify the specific surface structures on white blood cells. The lectin was isolated using affinity chromatography where a complex (oligosaccharide) matrix was used. Agglutination, binding and sugar inhibition assays confirmed the isolated protein was a lectin. The lectin was found in low amounts (up to 5% of the total protein content) within the seeds of G.robusta. As a result of this low yield, the identification of the lectin by PAGE was difficult because the levels of protein were beyond the detection limit of the commercial staining reagents. The lectin was called the GR2 protein and was characterised as a monocot mannose binding lectin based on its sugar specificity for only mannose. A serine protease inhibitor was isolated from the seeds of G.robusta using two different chromatography methods, reverse phase HPLC (GR1.HPLC) and gel filtration chromatography (GR1.GF). Ion exchange chromatography was used to initially separate the proteins in the crude extract and the fraction containing the GR1 protein was further purified using reverse phase HPLC (GR1.HPLC). N-terminal sequencing results of the GR1.HPLC protein, showed evidence of proteolytic cleavage during the extraction process, which lead to the second purification method being established. Protease inhibitors were added to the buffers prior to being purified by gel filtration chromatography, which resulted in the GR1 protein being isolated from the crude extract without the presence of the contaminating protein. Mass spectroscopy identified the molecular weight of the GR1 protein to be 6669Da and the full amino acid sequence was derived by cDNA techniques. Sequence alignment studies of the GR1 protein showed significant similarities with the Bowman-Birk inhibitor. The positioning of the cysteine residues were conserved throughout the Bowman-Birk superfamily, however these residues were not conserved within the GR1 protein. Competitive inhibition assays on the GR1 protein revealed the protein could inhibit both trypsin and chymotrypsin at similar levels to that seen for the Bowman-Birk inhibitor. Therefore, the GR1 protein was characterised as a member of the Bowman-Birk superfamily of serine protease inhibitors. The three-dimensional structure of the GR1 protein was determined using two-dimensional NMR spectroscopy. Computer programs such as XEASY, DYANA and SYBYL® were used to tabulate the information taken from the 2D experiments, generate structures and minimise these structures respectively. The solution structure of the GR1 protein was found to contain a region of antiparallel β-sheet structure that corresponded to the trypsin binding site and the remainder of the protein consisted of loops and turns that were held together by disulfide bridges (the chymotrypsin-binding region). Structural similarities between the GR1 protein and the Bowman-Birk inhibitor existed only in the trypsin-binding site of the Bowman-Birk inhibitor. The GR1 protein is the first member of the Proteaceae family to be characterised as a Bowman-Birk inhibitor. This thesis outlines the isolation and biochemical characterisation of the two proteins found within Grevillea robusta and also describes the steps involved and results obtained in determining the three-dimensional structure of the GR1 protein.