Plants are colonised by a vast amount of bacteria which are found in parts such as seeds, roots, leaves and fruits while fewer are found on blossoms, stems and vascular tissue. These different parts of plants make up distinct micro ecosystems which may result in different bacterial species (endophytes) colonizing these ecosystems. Such interactions could be for life or only a short period of time and may cause no significant damage or they could be latent pathogens. Isolations of both Gram negative and Gram positive bacteria have been made from an extensive range of plant species and include bacterial genera from the following groups: Firmicutes, Actinobacteria, á Proteobacteria, â Proteobacteria, and ã Proteobacteria. The focus of this study was the endophytic bacterial community of resistant, healthy and diseased Eucalyptus nitens clones, the latter of which showed symptoms of bacterial blight and die back previously described as caused by Pantoea ananatis. The endophytic bacteria of these sampled clones were studied using culturing dependent and independent methods. The focus was on the Enterobacteriaceae in order to determine whether P.ananatis is present as an endophyte of these clones. To obtain the isolates, standard culturing techniques were used, followed by sequence identification of the 16S rRNA as well as two housekeeping genes, rpoB and gyrB. Results obtained from the culturing study were compared to results obtained from a PCR-DGGE study of the same samples. Although no conclusion could be drawn as to which organism present caused the disease symptoms on the susceptible clones, it was seen that Enterobacter and Pantoea, were the most frequently isolated in both of the studies from all clones sampled. This implies that they are present as endophytes in the E.nitens clones, together with Pseudomonas and Bacillus as suggested by the DGGE study. Copyright / Dissertation (MSc)--University of Pretoria, 2012. / Microbiology and Plant Pathology / unrestricted

Eucalypts are a major forest resource globally and the area of eucalypt plantations for pulp and paper production is expanding rapidly in Australia. Consequently, there is an increasing need to breed eucalypts with improved wood properties. Since many high value wood traits are under strong genetic control, identification of DNA markers linked to these traits will have application in breeding programs. In recent years there has been a shift in marker strategy away from QTL mapping in pedigrees to association studies in unrelated populations. In the latter approach, single nucleotide polymorphisms (SNPs) in candidate genes are screened to identify SNPs that significantly associate with wood traits. Significant SNPs could be used for marker-assisted selection (MAS) in breeding programs. The objectives of this study were to identify candidate genes that may influence pulp yield in eucalypts and to identify SNP variants in those genes that associate with superior wood and pulp traits. / Approximately 300 trees from a full-sib Eucalyptus nitens progeny derived from a wide intra specific cross were used for gene discovery. DNA microarrays containing ~5800 young xylem of cDNAs Eucalyptus grandis were screened with probes synthesised from RNA isolated from trees with either high or low pulp yield. Forty-six transcripts were differentially regulated, of which 27 were more abundant in high pulp trees and 19 were more abundant in low pulp trees. All differentially expressed cDNAs were partially sequenced and searched against existing gene databases. Six genes were selected as putative pulp yield candidate genes based on their significant similarity to genes with known function and were named EgrCesA3 (cellulose synthase), EgrNAM1 (NAM family protein), EgrXET (xyloglucan endotransglycosylase), EgrGalk (galactokinase), EgrHB1 (class III homeodomain leucine zipper protein) and EgrZnf1 (C3HC4 type zinc finger protein). / Real-Time PCR was carried out on selected genes to confirm the accuracy of the microarray results. Full length cDNAs were obtained for EgrCesA3, EgrHB1 and EgrZnf1 and the candidate genes were partially characterised. An additional candidate gene, the novel gene EgrPAAPA, was selected based on previous research due to its high expression in the cambium and its expression in eucalypt branches. EgrPAAPA was cloned by screening an E. grandis cDNA library and fully sequenced. The full length EgrPAAPA encodes a short 172 amino acid protein rich in alanine, glutamic acid and proline residues. The EgrPAAPA protein appears to be a hydroxyproline-rich glycoprotein (HRGP) and the repetitive ‘PAAPA’ motif suggests that it might play a structural role in cell wall development. Southern blot analysis revealed that E. grandis has a single copy of the EgrPAAPA gene and northern blot analysis revealed that EgrPAAPA is most strongly expressed in xylem tissues. / Allelic variation in EnCesA3, EnNAM1, EnPAAPA and EnHB1 was examined by sequencing each gene in 16 to 24 unrelated E. nitens individuals. SNPs were identified by sequence analysis and patterns of nucleotide diversity, linkage disequilibrium and the selection of suitable polymorphisms were estimated. A moderate level of nucleotide diversity (θw = 0.0056 and π = 0.0039) was observed and linkage disequilibrium was generally low, extending only a few hundred base pairs in each gene. Negative selection has been operating in EnHB1. Selected TagSNPs from EnNAM1, EnHB1 and EnPAAPA were genotyped across 300 unrelated E. nitens trees which had been phenotyped for six wood quality traits including pulp yield, cellulose, lignin, Klason lignin, microfibril angle (MFA) and density. Five highly significant genetic associations (p<0.01) were detected between several SNPs in EnHB1 and all wood quality traits except density. A significant association was also found between EnPAAPA and MFA (p<0.05). No significant associations were found with any of the EnNAM1 SNPs. The strong genetic associations between SNPs in EnHB1 and a range of wood traits is consistent with this gene’s known role as a transcription factor controlling vascular development. Validation of these associations in different populations will be necessary in order to confirm these results. Alternatively, QTL mapping can be performed in order to confirm whether QTL for wood property traits can be detected at the EnHB1 and EnPAAPA loci.

Memoria para optar al Título Profesional de Ingeniero Forestal

Made available in DSpace on 2015-03-26T14:01:13Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1338814 bytes, checksum: 7845932ffe3998db89f67562073ff35a (MD5) Previous issue date: 2011-07-19 / Foi analisada, experimentalmente, o efeito da adição de antraquinona ao processo kraft convencional para produção de celulose de mistura de Eucalyptus globulus e Eucalyptus nitens. Foram analisados aumentos da velocidade da reação de deslignificação e estabilidade dos carboidratos, utilizando condições industriais de cozimento batch da Celulosa Arauco y Constitucion SA, Planta Arauco. Para alcançar kappa constante (15 e 17) foram modificados os tempos de cozimentos e foram utilizadas diferentes dosagens de antraquinona. Foram estabelecidas relações lineares entre a dosagem de antraquinona e o rendimento depurado, o teor de rejeitos, o alcali residual e o teor de sólidos no licor residual. Para a dosagem máxima de AQ utilizada (0,10%) foram obtidos, para ambos os níveis de kappa, aumentos de 1,95% e 1,32% nos rendimentos depurados, economias de 14,2% e 50,4% nas cargas de álcali e decréscimos nos tempos de cozimento de 13 e 11 minutos para kappa 15 e 17, respectivamente. / En la presente disertación se verificó de manera experimental el efecto de la adición de Antraquinona al proceso de cocción batch convencional para una mezcla industrial de Eucalyptus globulus E. nitens.; en lo que respecta a su capacidad de aumentar la velocidad de la reacción de deslignificación y proporcionar estabilidad a las hemicelulosas y celulosas de la fibra. Lo anterior se logró con experiencias de laboratorio fijando las condiciones de cocción de acuerdo a la operación de los digestores batch de Celulosa Arauco y Constitución S.A., Planta Arauco. Donde para obtener un kappa constante (15 y 17) se modificó el Factor H mediante el tiempo de cocción, en distintos niveles de carga de Antraquinona. Se pudo establecer una relación lineal entre la dosificación de Antraquinona y los parámetros estudiados: rendimiento clasificado, rechazos, álcali residual, tiempo de cocción y sólidos en el licor negro. Particularmente, para la máxima dosificación de AQ utilizada en este estudio (0,1% bms) se obtuvo para ambos niveles de Kappa respecto al blanco o testigo- un aumento de 1.95% y 1.32 de rendimiento clasificado, un ahorro de álcali de 14.2% y 50.4% y una disminución de tiempo de cocción de 13 min y 11 min para Kappa 15 y 17, respectivamente.

This dissertation examined the impact of species, physiological age and spacing on tree form and branching at a Nelder experiment located near Rolleston, Canterbury. Two species were compared, Pinus radiata and Eucalyptus nitens, at a range of stockings from 271 stems/ha to 40,466 stems/ha. Within the P. radiata, two different physiological ages were compared. Stocking and species significantly affected (p-value <0.05) tree height, diameter at breast height (DBH), crown depth, branch mortality, branch angle, branch size and internode length. Only stocking was statistically significant for crown width, and height from the ground was also statistically significant for branch angle and branch mortality. DBH, crown width, crown depth, branch size and branch survival decreased with increasing stocking for both species. Branch angle and average internode length increased as stocking increased for both species, and branch angle and average internode length also increased as you moved away from the base of the tree. DBH, average internode length and branch size were significantly larger for P. radiata across all stockings, however branch mortality and branch angle were significantly larger for E. nitens. Physiological age was not statistically significant for any aspects of tree form or branching examined in this study.

Biomass waste has been recognised as a promising, renewable source for future transport fuels. With 1.7 million hectares of pine plantation forests and 12 million cubic meters of annual residue produced by sawmills and the pulp and paper industries, New Zealand presents a prime location where utilisation of these resources can take the next step towards creating a more environmentally friendly future. In this research, the process of fast pyrolysis was investigated using a laboratoryscale, nitrogen-blown fluidised bed pyrolyser at CRL Energy. This equipment can process 1–1.5 kg/h of woody biomass in a temperature range of 450–550°C. The purpose of this rig was to determine the impact of various processing parameters on bio-oil yields. Next, the pyrolysis liquids (bio-oil and tar) were processed downstream into bio-bitumen. Pyrolysis experiments were carried out on Pinus Radiata and Eucalyptus Nitens residue sawdust from sawmills and bark feedstock. The properties of the collected products, including pyrolysis liquids (bio-oil and tar), gas and solid bio-chars, were measured under different operational conditions. Further analysis was also performed to determine pH, volatile content, chemical composition and calorific values of the products. The ultimate goal for this project was to develop a feasible, advanced fast-pyrolysis system for a bio-bitumen production plant using various biomass feedstocks. Additionally, a design for a bio-bitumen production plant was developed, and techno-economic analysis was conducted on a number of plant production yield cases and bio-bitumen manufacture ratios.

The influence of initial stocking and physiologically aged cuttings (taken from 1-year- old and 5-year-old parents) on corewood modulus of elasticity (MOE) in 6-year-old Pinus radiata D. Don was studied in a Nelder-design experiment in Rolleston, Canterbury. In the same experiment, the influence of initial stocking on MOE in 5-year- old Eucalyptus nitens was also investigated. The study incorporated 19 different stocking levels ranging from 207 to 40,446 stems/ha. Green dynamic modulus of elasticity was assessed in standing trees using the TreeTap stress-wave method over the lower part of the stem (0.3 – 1.9m) for 151 P. radiata trees and 115 E. nitens trees. The interaction between species and stocking significantly influenced MOE (P<0.001). MOE of P. radiata increased by 55% (or 3.9 GPa) between 271 and 40,466 stems/ha, and by 41% (or 2.2 GPa) between 271 and 4370 stems/ha. MOE of E. nitens was also influenced by stocking but the slope was significantly lower indicating that the effect of stocking was less pronounced than for P. radiata. Over the usual range of stockings for E. nitens there was an insignificant relationship between stocking and stiffness (P=0.335). Trees were also assessed for DBH, height, and stem slenderness (height/ DBH). None of these latter variables had a significant influence on MOE after the effects of stocking and species were accounted for. No effects of physiological age of cuttings were detected in this study. The findings of this study highlight the importance of stocking as a tool that forest managers can utilize to regulate corewood stiffness in P. radiata trees. These results also suggest that for E. nitens, where wood stiffness is a priority, forest managers could reduce establishment costs by planting at much lower initial stockings. This study also highlights the superior stiffness of E. nitens in direct comparison with P. radiata, with many trees in the experiment already exceeding stiffness thresholds for structural timber in New Zealand.

Mixed-species plantations of eucalypts and acacias have the potential to improve stand productivity over that of respective monocultures through the facilitative effect of nitrogen-fixation by acacias, and increased resource capture through above- and belowground stratification. However, growth in mixed-species plantations may not be improved compared to that of monocultures when competitive interactions outweigh the effects of improved nutrient availability and resource capture. Careful selection of sites and species is therefore critical to successfully improving stand productivity using mixed-species plantations. This study set out to examine some of the processes and interactions that occur in mixed-species plantations, and the effect nutrient and water availability can have on the growth of mixtures. In three out of four mixed-species field trials examined in this study, growth was not increased in mixtures compared to monocultures. However, in the fourth field trial, heights, diameters, stand volume and aboveground biomass were higher in mixtures of E. globulus and A. mearnsii from 3-4 years after planting. The range in outcomes from mixing species in these four trials shows that a fundamental understanding of the underlying processes is required to enable a greater predictive capacity for the circumstances under which mixtures will be successful. Therefore the growth dynamics, processes and interactions were examined in the mixtures of E. globulus and A. mearnsii. The difference in productivity between mixtures and monocultures in this trial increased with time up to age 11 years, when 1:1 mixtures contained twice the aboveground biomass of E. globulus monocultures. The positive growth response of trees in mixture compared to monocultures was the result of accelerated rates of nutrient cycling, a shift in C allocation and reductions in light competition through canopy stratification. Nitrogen contents of foliage and soil clearly showed that A. mearnsii influenced the N dynamics in this trial. If these changes in N contents were due to N fixation by A. mearnsii, then about 51 and 86 kg N ha-1 yr-1 was fixed in the 1:1 mixtures and A. mearnsii monocultures, respectively. Nitrogen fixation was also examined using the natural abundance method. The delta15N values of foliage collected at 10 years were grouped according to the mycorrhizal status of the host plant. Therefore the discrimination of 15N during transfer from mycorrhizae to the host plant appeared to vary with mycorrhizal status, and the natural abundance of 15N was not used to quantify N fixation. Rates of N and P cycling in litterfall were significantly higher in stands containing at least 25% A. mearnsii (more than 31 kg N ha-1 yr-1 and more than 0.68 kg P ha-1 yr-1) compared to E. globulus monocultures (24 kg N ha-1 yr-1 and 0.45 kg P ha-1 yr-1). Rates of litter decomposition and N and P release were about twice as high in 1:1 mixtures compared to E. globulus monocultures and were even higher in A. mearnsii monocultures. It is therefore important to select N-fixing species that are capable of cycling nutrients quickly between the plant and soil, and that have readily decomposable litter. The total belowground C allocation was not significantly different between mixtures and monocultures (14 to 16 Mg C ha-1 yr-1). However, since aboveground net primary production was greater in 1:1 mixtures, the changes in nutrient availability appears to have increased total productivity (both above- and belowground), and reduced the proportion of C allocated belowground in mixtures compared to E. globulus monocultures. In a pot trial containing mixtures of E. globulus and A. mearnsii both species grew larger in mixture than in monoculture at low N levels, and mixtures were more productive than monocultures. However, at high N levels, E. globulus suppressed A. mearnsii and mixtures were less productive than E. globulus monocultures. Similar effects were found for high and low levels of P. Therefore resource availability can have a strong influence on the interactions and growth of mixtures. The productivity of mixtures may only be increased on sites where the resource for which competition is reduced in mixture is a major limiting growth resource. For example, if N is not a limiting growth factor then an increase in N availability from N-fixation may not increase growth, and the N-fixing species may compete for other resources such as soil P, moisture or light. This study has shown that mixtures containing a N-fixing trees and a non-N-fixing trees can be more productive than monocultures, but that this increase in productivity will only occur on certain sites. Examination of the growth, interactions and processes that occurred in mixtures in this study provide useful information that can aid the selection of species combinations and sites.

The increased soil organic phosphorus (P) mineralization observed under trees compared to pasture has been attributed to the contrasting mycorrhizal associations of the two systems but to date, little work has been conducted comparing P dynamics under different tree species with contrasting mycorrhizal associations. This study investigated rhizosphere P dynamics and P acquisition of three tree species with contrasting mycorrhizal associations (ectomycorrhizal Pinus radiata, arbuscular mycorrhizal Cupressus macrocarpa and tripartite Ecualyptus nitens) using a combination of field and controlled environment studies. Short-term field studies revealed greater acid phosphomonoesterase activity and therefore greater potential organic P mineralization under radiata pine and eucalypt compared with macrocarpa, which correlated with ectomycorrhizal colonization. Related field work revealed that the presence of pasture understory and litter had a significant impact on P dynamics. Pasture acts to increase phosphatase activity and microbial activity within the soil but with a litter layer, the increased activities are mainly above the soil in the overlying litter. A glasshouse study investigated the separate influence of mycorrhizal hyphae and roots and results showed that ectomycorrhizal fungi associated with radiata pine and eucalypt stimulated microbial activity and increased rhizosphere phosphatase activity with a concomitant decline in soil organic P. Macrocarpa and eucalypt roots exuded higher concentrations of low molecular weight organic anions compared with radiata pine which stimulated microbial activity, increased rhizosphere phosphatase activity and decreased soil organic P. A series of experiments were then conducted to further investigate the precise role of mycorrhizae in P dynamics and results showed that ectomycorrhizal colonization increased rhizosphere acid phosphomonoesterase to a greater extent than arbuscular mycorrhizae. Fluorescent staining revealed that the mycorrhizae associated with all three species had similar phosphatase production capacities. The ability of the three species to utilize organic P was investigated in two exhaustive pot experiments. Radiata pine was shown to be the best adapted to utilize organic P, especially in soils with high relative organic P contents. Eucalypt appeared to enhance soil organic P mineralization but did not take up the released inorganic P, while macrocarpa caused limited organic P mineralization compared with radiata pine and eucalypt. This work shows that ectomycorrhizal trees are best adapted to utilize organic P through increased rhizosphere phosphatase activity and stimulated microbial activity. Root exudation of LMWOAs is of higher relative importance to arbuscular mycorrhizal trees to increase microbial activity and consequently phosphatase activities when fewer hyphae are in the rhizosphere are present to produce them.