The objective of this study was to determine how final stocking influences tree growth and radial and longitudinal variation in wood properties at a Pinus radiata D. Don plantation located at one of the warmest forest sites in New Zealand, Forsyth Downs forest in Northland. This thesis addressed both the effect of stocking on stand basal area, height, diameter and branch diameter and the effect of stocking on wood properties microfibril angle (MFA), module of elasticity and density. Finally, how ring width influences wood properties and whether this variable accounts for the treatment effects was investigated. Stocking, height and ring number and all interactions between these variables significantly affected ring width. Ring width by itself was significant as a predictor of density, but when it was combined with other class level variables it was insignificant (i.e. does not account for treatment effects), and it did not add anything to a model with only class effects. There was a significant impact of ring number on density while ring width was insignificant in the same model. MFA was significantly affected by ring width, height and ring number in the tree, and all interactions, apart from the three way interaction, but not by stocking. Ring width was significant in the MFA model both by itself and when it was combined with other variables. Ring width accounted for the stocking effect. The best model of MOE included the class level effects of stocking, height and ring number within the tree, and all interactions between these variables, and ring width, as a continuous variable. While there was a significant effect between stockings this was relatively weak compared to the other main effects. Ring width largely accounted for the effect of stocking, but not that of ring number, or height.

Low embodied energy, ability to act as a carbon store and ease of recycling gives forest products an important role within a low carbon built environment. Almost 25 % of the coniferous resource within the South West of England is Douglas-fir, a species reputed for producing high quality timber. Despite this, the region is facing challenges in delivering the resources full potential, a contributing factor to which is a loss of knowledge regarding its quality. The aim of the work presented is to gain an improved understanding of the quality of Douglas-fir grown within the region, from the perspective of uses in structural applications, the factors which influence material quality and their interrelationships. Flexural modulus of elasticity, flexural and compressive strength were determined utilising small clear specimens derived from 1.3 and 8 m heights within 27 trees from six sites across the South West. Results showed a rise in the magnitude of properties with increasing cambial age, particularly so at younger ages. Differences in values were also recorded between stem heights and with rate of growth. These were however less than age related variations. Results compared favourably to those reported in other studies conducted on the species. Utilising SilviScan-3, anatomical properties including density, microfibril angle and cellular dimensions were measured. Significant variations were recorded with cambial age, and in some instances sampling height. The influence of growth rate on anatomical properties was small. Through statistical and composite modelling, microfibril angle was found to be strongly associated with changes in modulus of elasticity within juvenile wood. Within mature wood and for strength properties, density was the controlling factor. It was shown that a moderate proportion of variations in mechanical properties can be accounted for utilising visually identifiable wood characteristics. The new understanding that has been gained through this work presents opportunities for improved utilisation, the implementation of effective management practices and the development of more efficient visual grading techniques.

The heritability of microfibril angle (MFA) in loblolly pine, Pinus taeda L., and its genetic relationships with height, diameter, volume and specific gravity were examined in two progeny tests with known pedigrees. Significant general combining ability (GCA), specific combining ability (SCA), and SCA x block effects indicated that there are both additive and non-additive genetic influences on MFA. Individual-tree narrow-sense heritability estimates were variable, ranging from 0.17 for earlywood (ring) 4 MFA to 0.51 for earlywood (ring) 20 MFA. Genetic correlations between MFA, specific gravity and the growth traits were non-significant due to large estimated standard errors. Multiple-trait selection and breeding in a mainline and elite population tree improvement program were simulated using Excel and Simetar (Richardson 2001). The effects of four selection indices were examined in the mainline population and the effects of seven selection indices and four breeding strategies were examined in the elite population. In the mainline population, selection for increased growth caused decreased wood quality over time. However, it was possible to maintain the overall population mean MFA and mean specific gravity at levels present in the base population by implementing restricted selection indices. Likewise, selection for improved wood quality in the elite population resulted in decreased growth unless restricted selection indices or pulp indices derived from those of Lowe et al. (1999) were used. Correlated phenotypic responses to selection on indices using economic weights and heritabilities were dependent on breeding strategy. When a circular mating system (with parents randomly assigned to controlled-crosses) was used, the index trait with a higher economic weight was more influential in determining correlated responses in non-index traits than the index trait with a lower economic weight. However, when positive assortative mating was used, the index trait with a greater variance was more influential in determining correlated responses in non-index traits than the index trait with a lower variance regardless of economic weight.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第20450号 / 農博第2235号 / 新制||農||1050(附属図書館) / 学位論文||H29||N5071(農学部図書室) / 京都大学大学院農学研究科森林科学専攻 / (主査)教授 杉山 淳司, 教授 髙部 圭司, 教授 梅澤 俊明 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Fibrillins are modular, disulphide-rich glycoproteins that assemble into microfibrils in the extracellular matrix (ECM). These microfibrils are critical structural elements of many non-elastic and elastic connective tissues. They also regulate the availability of transforming growth factor-β signalling molecules in the ECM. Defects in microfibrils are associated with acquired and inherited connective tissue disorders. In particular, mutations in the human FBN1 gene, which encodes fibrillin-1, are associated with a spectrum of diseases, including Marfan syndrome (MFS). One of the proposed initial steps in microfibril assembly is the interaction between the N- and C-terminal regions of fibrillin monomers. The minimal regions of human fibrillin-1 required for an interaction in vitro were previously identified: the four N-terminal domains, from the fibrillin unique N-terminal (FUN) domain to the third epidermal growth factor-like (EGF) domain (FUN-EGF3), and the three C-terminal calcium-binding EGF-like (cbEGF) domains (cbEGF41-43). Here, fragments corresponding to these regions were produced and shown to interact in pull-down and surface plasmon resonance assays. In addition, the structure of the FUN-EGF3 fragment was determined using nuclear magnetic resonance spectroscopy. This showed the novel structure of the FUN domain and the interdomain interfaces in this region of fibrillin. Combining structural and sequence conservation data may help to identify regions of FUN-EGF3 important for binding to cbEGF41-43. Here, the interaction was probed by site-directed mutagenesis. However, substituting individual residues in FUN-EGF3 with alanine did not abrogate binding to cbEGF41-43. Three MFS-associated residue substitutions were also introduced into the FUN-EGF3 fragment. While they did not abolish the interaction with cbEGF41-43, they did cause misfolding. Two of these substitutions, N57D and W71R, also resulted in the defective secretion of a larger N-terminal fragment by fibroblast cells, suggesting a potential mechanism of disease pathogenesis. Although specific residues involved in the N-C interaction were not identified here, the FUN-EGF3 structure will be vital for understanding the molecular surfaces involved in microfibril assembly and growth factor binding.

Plant cell walls are complex structures that must not only constrain cellular turgor pressure but also allow for structural modification during the dynamic processes of cell division and anisotropic expansion. Cell walls are composed of highly glycosylated proteins and polysaccharides, including pectin, hemicellulose and cellulose. The primary cell wall polysaccharide is cellulose, a polymer composed of high molecular weight !- 1,4-glucan chains. Although cellulose is the most abundant biopolymer on Earth, there is still a lot to learn about its biosynthesis and regulation. This research began by applying a variety of analytical techniques in an attempt to understand differences in cell wall composition and cellulose structure within the plant body, between different plant species and as a result of acclimation by the plant to different environmental conditions. Next, a number of different Arabidopsis thaliana lines possessing mutations affecting cell wall biosynthesis were analyzed for changes in cellulose structure (crystallinity) and biomass saccharification efficiency. One of these mutants, isoxaben resistance1-2 (ixr1- 2), which contains a point mutation in the C-terminal transmembrane region (TMR) of cellulose synthase 3 (CESA3), exhibited a 34% lower biomass crystallinity index and a 151% improvement in saccharification efficiency relative to that of wild-type. The culmination of this research began with a chemical screen that identified the molecule quinoxyphen as a primary cell wall cellulose biosynthesis inhibitor. By forward genetics, a semi-dominant mutant showing strong resistance to quinoxyphen named aegeus was identified in A. thaliana and the resistance locus mapped to a point mutation in the TMR of CESA1. cesa1aegeus occurs in a similar location to that of cesa3ixr1-2, illustrating both subunit specificity and commonality of resistance locus. These drug resistant CESA TMR mutants are dwarfed and have aberrant cellulose deposition. High-resolution synchrotron X-ray diffraction and 13C solid-state nuclear magnetic resonance spectroscopy analysis of cellulose produced from cesa1aegeus, cesa3ixr1-2 and the double mutant shows a reduction in cellulose microfibril width and an increase in mobility of the interior glucan chains of the cellulose microfibril relative to wild-type. These data demonstrate the importance of the TMR region of CESA1 and CESA3 for the arrangement of glucan chains into a crystalline cellulose microfibril in primary cell walls.

This study approaches wood quality in young trees by very early screening – and consequent selection for propagation – on the basis of physical and mechanical properties. In chapter 1 corewood properties are reviewed and the importance and problems associated with early screening are discussed. Due to randomly distributed reaction wood in young trees it is advantageous to lean trees to avoid intermixing of the two wood types and minimise any uncertainty in the results. In chapter 2 physical and mechanical properties are described for opposite and compression wood in a population of Pinus radiata comprising of 50 families, at a young (<3 years) age. The dynamic stiffness was determined using the resonance acoustic technique. Density was measured using water displacement method, and longitudinal and volumetric shrinkage were measured from green to ~5% moisture content. The compression wood and opposite wood differ significantly in all the measured properties. Compression wood was characterised by high density and high longitudinal shrinkage. The mean stiffness of opposite wood was 3.0 GPa with a mean standard deviation of 0.39, and the mean longitudinal shrinkage of opposite wood was 0.99% with mean standard deviation of 0.31 across the samples examined. This variation in stiffness and longitudinal shrinkage in opposite wood can be exploited to screen for wood quality. The variation in stiffness and longitudinal shrinkage within a family was comparable to variation among families. In spite of large within site variability it was possible to distinguish between the worst and the best families in opposite wood at young age. In chapter 3 ranking of selected families of Pinus radiata was done based on microfibril angle, which is considered as the main factor influencing both stiffness and longitudinal shrinkage. The ranking was compared with ranking done using acoustic velocity which is more practical and fast method of screening trees. The mean MFA in opposite wood was 39° with a mean standard deviation of 3.7 and in compression wood the mean MFA was 44° with a mean standard deviation of 2.9. The variation in MFA in opposite wood offers opportunities to breed for trees with low MFA. A strong negative correlation (R=-0.68) between acoustic velocity squared and MFA in opposite wood suggested that the resonance technique can be used effectively to screen very young wood rather than using MFA. At high MFA, the cell wall matrix also plays an important role in determining the mechanical and physical properties of the wood. At present the chemical composition of wood samples is determined by wet chemical analysis, which is time consuming and laborious. Therefore, it is impractical to characterise large numbers of samples. Mechanical properties, particularly tanδ (dissipation of energy), which changes with temperature and frequency as the structure of the material changes at the molecular level, was studied using dynamic mechanical analysis (DMA). The idea was to assess if it can be used as a quality trait for tree screening instead of wet chemical analysis. Compression wood and opposite wood were characterised for storage modulus and tanδ at constant moisture content. In practice the instrument used, TA instrument Q800, was unable to provide the desired range of temperature and humidity so no glass transition at 9% moisture content in the temperature range of 10°C to 85°C at 1 and 10 Hz frequency was observed that might be attributed to the hemicelluloses (or lignin). In spite of the huge difference in chemical composition of opposite and compression wood, the difference in their mean tanδ at 25°C and 1 Hz values was just 7%. The positive correlation between MFA and tanδ in opposite wood suggested that MFA also plays a role in the dissipation of energy. The strong relationship between storage modulus and dynamic modulus (R=0.74) again justifies the reliability of resonance technique to screen young wood for stiffness. Concurrently eighty seven, two-year-old leant Eucalyptus regnans were studied for growth strains along with other physical and mechanical properties, independently in tension and opposite wood. The leant trees in Eucalyptus regnans vary in their average growth strain. Strong correlation between measured and calculated strain (R=0.93) suggests that the quick split method can be used to screen large populations for growth stresses. Tension wood was characterised by high density and was three times stiffer than opposite wood and twice as high in volumetric shrinkage. The high longitudinal shrinkage in opposite wood could be due to comparatively high MFAs in opposite wood of the young trees. There was no correlation between growth strain values and other measured properties in opposite wood. It is possible to screen for growth strain at age two, without any adverse effect on stiffness and shrinkage properties.

Cellulose nanowhiskers reinforced poly(vinyl alcohol)(PVA) nanofiber web is successfully fabricated using electrospinning technique and the mechanical properties of the single electrospun fiber are measured using nanoindentation method. The morphology and mechanical properties of highly aligned electrospun fiber webs are investigated. It is found that the modulus and tensile strength of aligned webs are higher than those of isotropic electrospun fiber webs. Experimental results are compared with a longitudinal Halpin-Tsai model. Ice-templated(IT) cellulose microfibril porous foams are successfully fabricated via unidirectional freezing methods. The morphology and growth mechanism of IT surfaces are investigated successfully using cellulose microfibrils and hydrophillic substrates. By controlling the temperature gradient between cellulose microfibril suspensions and secondary freezing mediums, various surface structures including honey-comb like structures, ellipse-shape channel strcutures, fully developed multichannel structures are obtained. For the honey-comb like patterned surface, high contact angles are observed. On the other hand, for the layered patterned surface, anisotropic wetting properties are observed.

Orientador: Cláudio Chrysóstomo Werneck / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-16T09:01:31Z (GMT). No. of bitstreams: 1 Machado_Denise_M.pdf: 1404386 bytes, checksum: e811cae8e94f35d5b473fe2881261fd8 (MD5) Previous issue date: 2010 / Resumo: Nos últimos anos, o principal objetivo de nosso laboratório é esclarecer a participação da GlicoProteína Associada a Microfibrila-1 (MAGP-1) na formação de trombos no modelo de trombose arterial fotoquímica. Dados indicam que a MAGP-1 não interage diretamente com as plaquetas, importante componente na trombose arterial, mas que, provavelmente, a MAGP-1 esteja interagindo com uma molécula que tenha participação neste processo. Neste sentido, a interação da MAGP-1 com moléculas importantes na formação do trombo foi estudada. A MAGP-1 tem a capacidade de interagir com o fator de von Willebrand, bem como com fibrinogênio e fibronectina. Além disto, foi verificado que a injeção da MAGP-1 recombinante em camundongos deficientes em MAGP-1 é capaz de reestabelecer o tempo normal de formação de trombos nestes animais. Considerando a ausência da MAGP-1 e seu efeito na formação de trombos, foi questionado se a morfologia dos trombos obtidos em camundongos deficientes em MAGP-1 era diferente dos trombos obtidos em camundongos selvagens. Dados iniciais sugeriam que os trombos de camundongos deficientes em MAGP-1 apresentavam ultraestrutura diferente, onde um número maior de plaquetas apresentavam aparentemente, os seus grânulos e corpos densos intactos, sugerindo uma ativação ineficiente na ausência de MAGP-1. No presente trabalho, além de análise ultraestrutural mais detalhada, foram feitos estudos para determinar qual a região da MAGP-1 é responsável pela atividade trombogênica. Neste sentido, proteínas mutadas, truncadas e peptídeos derivados da MAGP-1 foram obtidos e então injetados nos camundongos selvagens e deficientes em MAGP-1. Como resultados, pudemos observar que os trombos dos camundongos selvagens bem como dos camundongos deficientes em MAGP-1 apresentam basicamente a mesma morfologia, levando em consideração as técnicas utilizadas, microscopia eletrônica de transmissão e varredura. Em relação ao mapeamento dos domínios, quando utilizada a molécula inteira, seja de camundongo, seja bovina, o tempo normal de oclusão foi reestabelecido. A injeção da forma truncada da região carboxiterminal foi suficiente para obter este mesmo resultado que está principalmente relacionado a um peptídeo também determinado. O mecanismo pelo qual este peptídeo atua normalizando o tempo de oclusão normal ainda não é conhecido e será objeto de estudos futuros / Abstract: In the last years, the main purpose of our laboratory has been to clarify the participation of the Microfibril-Associated GlycoProtein-1 (MAGP-1) in forming thrombus in photochemically-induced artery thrombosis model. It was considered that MAGP-1 does not interact directly with platelets, important component in artery thrombosis and probably MAGP-1 is interacting with a molecule which has participation in this process. In this sense, the interaction of the MAGP-1 with important molecules in the thrombus formation was studied. The MAGP-1 has the ability to interact with von Willebrand factor, as well as with fibrinogen and fibronectin. In addition, it was verified that the injection of recombinant MAGP-1 in MAGP-1-deficient mice was able to restores normal time of thrombus formation in these animals. Considering the MAGP-1 absence and its effect on the thrombi formation, it was questioned whether thrombi morphology obtained from MAGP-1-deficient mice was different from thrombi obtained from wild-type mice. Initial data suggested that the thrombi from MAGP-1-deficient mice had different structure, where platelets apparently showed their intact granules and dense bodies, suggesting an inefficient activation in the MAGP-1 absence. In this work, besides the ultra-structural analysis, more detailed studies have been made to determine which region of the MAGP-1 is responsible for the thrombogenic activity. In this sense, truncated proteins, mutated proteins and peptides, were obtained and then injected into MAGP-1-deficient mice to verify their activities. As result, we noticed that the thrombi have similar structure taking into consideration used techniques, scanning electron microscopy and transmission electron microscopy. Furthermore, in relation to MAGP-1 mapping domains studies when we inject recombinant full-length molecule either mice or bovine, the normal time occlusion was restored. The injection of carboxy-terminal region was enough to get this same result what is mainly related to an also determined peptide. Mechanisms involved in this process are unknown and will be our aim in the future studies / Mestrado / Bioquimica / Mestre em Biologia Funcional e Molecular

The content of this study is focused on the influence of the structure of wood at microscopic and submicroscopic level on its mechanical properties. The wood cell wall consists of several layers, the dominant layer being layer S2, which occupies up to 80 % of the total thickness of the wood cell wall. Unique feature of this layer is that cellulose microfibrils placed in this layer are highly aligned and spirally wound around the cell axis. The inclination of these microfibrils is called microfibril angle (MFA) and is the key feature that affects mechanical properties of wood and its shrinkage. In theoretical part of this thesis methods for measuring microfibril angle are described. A method for measuring mechanical properties of the wood cell wall called nanoindentation is discussed in detail. In the practical part of this thesis, microfibril angle is measured by means of polarized light microscopy and mechanical properties of wood cell wall is determined by means of nanoindentation.