Pinus radiata D. Don is an economically important plantation species to New Zealand that is susceptible to the wood quality flaw 'intra-ring checking'. Intra-ring checking is a term used to describe radial fractures that can occur in the earlywood portion of a growth ring, altering the appearance and resilience of the wood, thereby decreasing its economic value. This thesis presents a study that was part of a broad, ongoing collaborative investigation directed at understanding wood quality issues, with the long term goal of enhancement of future radiata pine crops. These investigations are funded by the Wood Quality Initiative Ltd., and involve basic science, field trials and engineering studies related to intra-ring checking. Specifically, the present study was designed to establish the effects of the mineral nutrients boron, calcium and magnesium on wood formation, to determine whether they are associated with intra-ring checking. This research was carried out in three stages. Firstly, the ultra-structural and biochemical properties of wood with intra-ring checking were examined to determine if specific features of the cell wall were associated with the incidence of intra-ring checks. Electron microscopy techniques revealed that the CML/S1 region of the cell wall often showed a decrease in CML lignin staining and S1 striations in wood with intra-ring checks. However, Klason and acetyl bromide assays did not show a change in lignin content. In order to understand how changes in the CML/S1 region of the cell wall may occur, methods were required that would allow for the observation of wood formation in a controlled environment. In the second stage of this study, an organ culture technique was successfully developed to allow for the growth of radiata pine cambial tissue, sandwiched between phloem and xylem, on a defined nutrient medium. This nutrient medium was manipulated, using ion-binding resins, to control the amount of boron, calcium and magnesium available to the growing tissues, to determine if variations in wood formation could be induced. In the final stage of this research, an extensive comparative examination of different techniques that could be used for the observation and measurement of selected wood properties was undertaken, in order to determine the efficacy of the organ cultures to study wood formation in an altered nutrient environment. Wood properties were examined for various stages of xylogenesis, beginning with cell division and expansion, followed by cell wall deposition, and lastly with the onset of lignification in order to define the success of the culture technique. Electron microscopy investigations suggested that in the presence of very little boron the CML/S1 wall showed darker striation deposits, while an increase in calcium availability, resulted in a more defined CML/S1/S2 wall region compared to the controls. Further examination of the cell walls suggested that pectin esterification and possibly lignification could also be increased by limited boron availability. However, in many of the observed and measured parameters of wood properties, a great deal of complex 'between-tree' and 'within-culture' variation was observed. The results show that elucidation of the association between nutrient availability and the incidence of intra-ring checking can not be established from this organ culture study. In a concurrent study, the preliminary investigation of arabinogalactan-proteins (AGPs) in radiata pine was undertaken. Radiata pine AGPs were positioned in the compound middle lamella of xylem cells, suggesting potential roles in cell-cell adhesion or cell-cell signalling. For the first time, radiata pine AGPs were isolated and characterized in terms of their protein and carbohydrate composition, both of which yielded features typical of AGPs in other plant species. Unique to radiata pine AGPs was the presence of a large proportion of 5-linked arabinose. While the precise function(s) of AGPs are unknown, the results obtained in this research have established a basis for further investigation into the potential for their involvement in wood formation. Overall, new tools have been established to facilitate future research on radiata pine, a commercially important species, and novel results have been obtained concerning the mechanisms of wood formation therein.

Pinus radiata D. Don is an economically important plantation species to New Zealand that is susceptible to the wood quality flaw 'intra-ring checking'. Intra-ring checking is a term used to describe radial fractures that can occur in the earlywood portion of a growth ring, altering the appearance and resilience of the wood, thereby decreasing its economic value. This thesis presents a study that was part of a broad, ongoing collaborative investigation directed at understanding wood quality issues, with the long term goal of enhancement of future radiata pine crops. These investigations are funded by the Wood Quality Initiative Ltd., and involve basic science, field trials and engineering studies related to intra-ring checking. Specifically, the present study was designed to establish the effects of the mineral nutrients boron, calcium and magnesium on wood formation, to determine whether they are associated with intra-ring checking. This research was carried out in three stages. Firstly, the ultra-structural and biochemical properties of wood with intra-ring checking were examined to determine if specific features of the cell wall were associated with the incidence of intra-ring checks. Electron microscopy techniques revealed that the CML/S1 region of the cell wall often showed a decrease in CML lignin staining and S1 striations in wood with intra-ring checks. However, Klason and acetyl bromide assays did not show a change in lignin content. In order to understand how changes in the CML/S1 region of the cell wall may occur, methods were required that would allow for the observation of wood formation in a controlled environment. In the second stage of this study, an organ culture technique was successfully developed to allow for the growth of radiata pine cambial tissue, sandwiched between phloem and xylem, on a defined nutrient medium. This nutrient medium was manipulated, using ion-binding resins, to control the amount of boron, calcium and magnesium available to the growing tissues, to determine if variations in wood formation could be induced. In the final stage of this research, an extensive comparative examination of different techniques that could be used for the observation and measurement of selected wood properties was undertaken, in order to determine the efficacy of the organ cultures to study wood formation in an altered nutrient environment. Wood properties were examined for various stages of xylogenesis, beginning with cell division and expansion, followed by cell wall deposition, and lastly with the onset of lignification in order to define the success of the culture technique. Electron microscopy investigations suggested that in the presence of very little boron the CML/S1 wall showed darker striation deposits, while an increase in calcium availability, resulted in a more defined CML/S1/S2 wall region compared to the controls. Further examination of the cell walls suggested that pectin esterification and possibly lignification could also be increased by limited boron availability. However, in many of the observed and measured parameters of wood properties, a great deal of complex 'between-tree' and 'within-culture' variation was observed. The results show that elucidation of the association between nutrient availability and the incidence of intra-ring checking can not be established from this organ culture study. In a concurrent study, the preliminary investigation of arabinogalactan-proteins (AGPs) in radiata pine was undertaken. Radiata pine AGPs were positioned in the compound middle lamella of xylem cells, suggesting potential roles in cell-cell adhesion or cell-cell signalling. For the first time, radiata pine AGPs were isolated and characterized in terms of their protein and carbohydrate composition, both of which yielded features typical of AGPs in other plant species. Unique to radiata pine AGPs was the presence of a large proportion of 5-linked arabinose. While the precise function(s) of AGPs are unknown, the results obtained in this research have established a basis for further investigation into the potential for their involvement in wood formation. Overall, new tools have been established to facilitate future research on radiata pine, a commercially important species, and novel results have been obtained concerning the mechanisms of wood formation therein.

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No description available.

Les gommes d’Acacia (AG, E414EC) sont largement utilisées par ses propriétés stabilisantes, émulsifiantes et filmogènes. Le but de ce projet de thèse consiste à caractériser les propriétés interfaciales et émulsifiantes des gommes d’Acacia. Les études des propriétés interfaciales liquide-liquide ont confirmé que l’A. senegal permettait une plus grande diminution de la tension interfaciale et une formation plus rapide de films à l’interface. Ceci est en accord avec sa teneur plus élevée en AGP de masse molaire élevée riche en protéine, la meilleure accessibilité de la partie protéique et la plus grande flexibilité moléculaire par rapport à A. seyal. De même, ces avantages biochimiques et structuraux de l’A. senegal se sont avérés être impliqué dans les propriétés émulsifiantes des gommes. Les résultats d'une approche innovante visant à contrôler la teneur en AGP de masse molaire élevée riche en protéines dans le milieu et la concentration totale de gomme par le mélange de deux fractions bien caractérisées isolées chez A. senegal ont confirmé la synergie fonctionnelle entre la quantité des AGP de masse molaire élevée riche en protéines et la concentration totale en gomme. De plus, en présence de quantités élevées d'AGP de masse molaire élevée riches en protéines, les émulsions étaient stables contre la floculation/coalescence en raison de la forte teneur en protéines permettant une répulsion électrostatique entre les gouttelettes et la viscosité apparente élevée du milieu. Celles-ci étaient en accord lorsque les propriétés émulsifiantes de A. senegal et A. seyal ont été comparées. En effet, A. senegal contient une plus grande quantité d'AGP de masse molaire élevée riches en protéines et d'acides uroniques, et sa dispersion présente une viscosité apparente supérieure à celle d'A. seyal. En outre, lorsque les gommes d'Acacia étaient utilisées sous forme de films séchés, la grande teneur en AGP riches en protéines de masse molaire élevée et la bonne accessibilité des fragments protéiques d'A. senegal permettaient aux films d’avoir une surface homogène lisse avec des propriétés hydrophobes caractéristiques. En revanche, les films de A. seyal étaient irréguliers et constitués d'une organisation répétitive comme de nombreuses grosses particules uniformément réparties sur la surface par rapport au faible degré de ramification, une teneur élevée en arabinose favorisant les liaisons hydrogènes intra et inter moléculaires et une forte capacité d'hydratation chaînes polysaccharidiques de A. seyal. Selon les résultats, cette thèse apporte de nouvelles connaissances sur la relation entre la composition biochimique, les propriétés structurales et physico-chimiques, pour la première fois, dans les deux aspects de la gomme, à savoir sous forme de dispersion liquide et de films séchés.Mots clés : gomme d’Acacia, propriétés interfaciales et émulsifiantes, arabinogalactane-protéines, arômes / Acacia gums (AG, E414 EC) are widely used for its stabilizing, emulsifying and film-forming properties. The aim of this project is to characterize the interfacial and emulsifying properties of Acacia gums. The main results from liquid-liquid interfacial properties study confirmed that A. senegal showed a faster decrease of interfacial tension and a more rapid interfacial film formation. These were in agreement to its greater content of high molar mass-protein AGPs content, the more accessibility of proteinaceous moieties, and the higher molecular flexibility compared to A. seyal. In the same vein, these biochemical and structural advantages of A. senegal were found to be involved in the emulsifying properties of gums. The results from an innovative approach aiming to control high molar mass protein-rich AGPs content in bulk and the total concentration of gum by mixing two well characterised fractions isolated from A. senegal confirmed the functional synergism between the amount of high molar mass protein-rich AGPs and the total gum concentration. Moreover, in the presence of high molar mass protein-rich AGPs in high amount, the emulsion became stable to flocculation/coalescence due to the great protein content allowing electrostatic repulsion between droplets and the high bulk apparent viscosity. These were in agreement when the emulsifying properties of A. senegal and A. seyal were compared. Indeed, A. senegal containing a greater amount of high molar mass protein rich AGPs and uronic acids and its dispersion having a higher apparent viscosity than A. seyal allowing the former to form emulsion with a more stability. Besides, when Acacia gums were used in the form of dried film, the great content of high molar mass protein-rich AGPs and the good accessibility of proteinaceous moieties of A. senegal allowed the smooth homogeneous surface with a hydrophobic characteristic properties of A. senegal dried films. In contrast, A. seyal films was irregular and composed of a repetitive organization as numerous large particle uniformly distributed on surface in the relation to the low degree of branching, high arabinose content favoring intra and inter molecular hydrogen bonding and high hydration ability causing aggregation of polysaccharide chains of A. seyal. According to the results, this thesis brings new knowledge of the relationship between biochemical composition, structural and physicochemical properties, for the first time, across both aspects of gum, i.e. in the form of liquid dispersion and dried films.Keywords: Acacia gums, interfacial and emulsifying properties, arabinogalactan proteins, aroma compounds

La gomme d’Acacia est l’une des plus anciennes gommes naturelles dans le monde et la plus connue. Elle est définie comme l’exsudat gommeux produit par les arbres d’Acacia senegal et Acacia seyal. Les gommes d’Acacia sont composées d’arabinogalactanes protéines (AGPs), faiblement chargés, hyperbranchés avec une forte proportion de sucres (90%) et d’environ 1-3% de protéines et de minéraux. Malgré ses nombreuses applications industrielles, les connaissances sur ses propriétés volumétriques (hydrostatiques et hydrodynamiques) restent à améliorer. Ces propriétés peuvent être liées à la flexibilité et l’hydratation des molécules qui déterminent les propriétés fonctionnelles importantes comme les propriétés interfaciales. L'objectif de cette thèse est l’étude des propriétés volumétriques d’AGPs de la gomme d’Acacia. L’étude a été faite sur les principales variétés des gommes d’Acacia, A. senegal et A. seyal, ainsi que des fractions macromoléculaires d’A. senegal, obtenues par la chromatographie d'interaction hydrophobe et d'échange ionique. Les principaux résultats ont montré que les AGPs de gomme d’Acacia ont une structure semi flexible. De plus, des différences dans la flexibilité et l’hydratation entre les fractions d’AGPs ont été montré. Ces différences ont été expliquées par leurs différences en composition, polarité, masse molaire, forme et conformation. De plus, un comportement intermédiaire entre des protéines et des polysaccharides linéaires ont été montré. Finalement, un effet des agrégats d’AGPs sur les propriétés volumétriques a été mis en avance. / Acacia gum is the oldest and most widely known and used gum, it is a dried gummy exudate from the leaves and branches of the Acacia senegal and Acacia seyal trees. Acacia gums are weakly charged, amphiphilic hyperbranched arabinogalactan-proteins (AGPs). They are composed of about 90% polysaccharides and from 1-3% of proteins and minerals. In spite of the widely spread of industrial usage of A. gums, their volumetric properties (hydrostatic and hydrodynamic) have not been well studied. These properties have been linked to important properties such as flexibility and hydration of the molecule, which are related to important functional properties of A. gums (e. g. interfacial properties). The main objective of this PhD thesis was to study the volumetric properties of AGPs from Acacia gums exudates. For this effect, the main commercial species, A. senegal and A. seyal, and the macromolecular fractions of the former, obtained via hydrophobic interaction and ionic exchange chromatographies were studied. The main results showed that AGPs from Acacia gums have a semi-flexible structure. However, differences in their flexibility and hydration were seen among AGP fractions. These differences were explained based on their composition, polarity, molar mass, shape and conformation. Furthermore, an intermediate behavior between proteins and linear polysaccharides was evidenced. In addition, an effect of the presence of AGP based aggregates on the volumetric properties was seen.

No description available.

Arabinogalactan proteins (AGPs) are proteoglycans heavily substituted by arabinogalactan polysaccharides. These are composed of arabinose and galactose, and minor sugars such as glucuronic acid (GlcA), fucose and xylose. The arabinogalactan polysaccharides do not decorate classical AGPs exclusively, but they can also be found decorating a wide range of proteins. Arabinogalactan proteins have been implicated in many processes of plant development. Recently, AGPs were proposed to bind and store calcium at the plasma membrane. They are extracellular, and are localised mainly at the plasma membrane via a GPI-anchor. They can also be soluble in the apoplast. Their low abundance, chemical similarity and high functional redundancy have hindered their study. My strategy to overcome these difficulties was to study knock-out Arabidopsis thaliana plants of glycosyltransferases that transfer sugars specifically onto AG-polysaccharides. Glucuronic acid makes up about 10% of the arabinogalactan polysaccharide structure in Arabidopsis thaliana cell culture AGPs. Previously, the glucuronic acid transferase A TGLCA T14A, a member of the CAZy Glycosyl Transferase 14 family, was shown to transfer GlcA specifically onto AGPs, and knock-out Arabidopsis plants showed a 30% reduction in [Me]GlcA substitution in AGP-enriched preparations. However, no clear growth phenotype was observed. The characterisation of knock-out plants of other GT14 family members and combinations thereof is described here. Based on previous studies (Lamport and Várnai, 2013), I assayed in vitro the calcium binding capacity of AGP extracts from WT and knock-out plants. The results showed that AGP extracts from knock-out plants can hold less calcium than WT plants in vitro. A wide range of plant growth phenotypes were identified. Growth phenotypes can be explained by changes in the cytoskeleton and deficiencies in calcium signaling. Our evidence suggests links between structural deficiencies of extracellular proteoglycans to extracellular calcium and cytoskeleton. This research has the potential to create a new model system for the study of molecular mechanisms dependent on calcium that drive cell expansion, division and differentiation in plants.

Adhesion between the plasma membrane and the cell wall and the existence of the continuum of these two compartments is needed for signal transmission, e.g. under pathogen attack, during cell expansion and cell wall growth, or in response to environmental conditions. This adhesion is, in addition to the turgor, provided by physical connection of both compartments. One of the best-known examples of physical connection is found in the root system, in the Caspary strip region, where it is required to maintain apoplastic barriers of the root system, even under adverse conditions and consequent plasmolysis. There is little information about the physical interconnection and the participating macromolecules but there are candidates, which could participate in this interaction. The diploma thesis deals with arabinogalactan (AGP) proteins with fasciclin-like domain (namely FLA9 and FLA2). These proteins may play a role in the adhesion of plasma membrane and cell wall and may be involved in the growth regulation and development of the root system. Both genes are are relatively strongly expressed in the roots, especially in the elongation zone and in the cortex, including the endodermis (according to chip data). Also, to a smaller extent, the thesis deals with integrin-like At14A protein. The most interesting...