Sustaining the socioeconomic and ecological benefits of South African plantation forests is challenging. A more systematic and rapid forest inventory system is required by forest managers. This study investigates the utility of medium (ASTER 15 m) and high (IKONOS 1-4 m) spatial resolution satellite imageries in an effort to improve the remote capture of structural attributes of even-aged Eucalyptus plantations grown in the warm temperate climatic zone of southern KwaZulu-Natal, South Africa. The conversion of image data to surface reflectance is a pre-requisite for the establishment of relationships between satellite remote sensing data and ground collected forest structural data. In this study image-based atmospheric correction methods applied on ASTER and IKONOS imagery were evaluated for the purpose of retrieving surface reflectance of plantation forests. Multiple linear regression and canonical correlation analyses were used to develop models for the prediction of plantation forest structural attributes from ASTER data. Artificial neural networks and multiple linear regression were also used to develop models for the assessment of plantation forests structural attributes from IKONOS data. The plantation forest structural attributes considered in this study included: stems per hectare, diameter at breast height, mean tree height, basal area, and volume. In addition, location based stems per hectare were determined using high spatial resolution panchromatic IKONOS data where variable and fixed window sizes of local maxima were employed. The image-based dark object subtraction (DOS) model was better suited for atmospheric correction of ASTER and IKONOS imagery of the study area. The medium spatial resolution data were not amenable to estimating even-aged Eucalyptus forest structural attributes. It is still encouraging that up to 64 % of variation could be explained by using medium spatial resolution data. The results from high spatial resolution data showed a promising result where the ARMSE% values obtained for stems per hectare, diameter at breast height, tree height, basal area and volume are 7.9, 5.1, 5.8, 8.7 and 8.7, respectively. Results such as these bode well for the application of high spatial resolution imagery to forest structural assessment. The results from the location based estimation of stems per hectare illustrated that a variable window size approach developed in this study is highly accurate. The overall accuracy using a variable window size was 85% (RMSE of 189 trees per hectare). The overall findings presented in this study are encouraging and show that high spatial resolution imagery was successful in predicting even-aged Eucalyptus forest structural attributes in the warm temperate climates of South Africa, with acceptable accuracy. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2008.

Assessment of forest structural attributes has major implications in the management of forestry by providing information of ecological and economic importance. The traditional methods of assessment involve collecting data in the field and are regarded as labour-intensive and expensive. In plantation forestry, field campaigns are generally time consuming and costly, and may compromise profit maximisation. The introduction of lidar (light detection and ranging) remote sensing in forestry has shown promise to add value to the traditional field inventories mainly through large spatial coverages in a timely and cost-effective manner. Lidar remote sensing is an advanced system capable of acquiring information in both the vertical and horizontal dimensions at relatively high resolutions. Numerous studies have established that these qualities of lidar data are suited to estimating forest structural attributes at acceptably high accuracies. The generic approach in most studies is to use lidar data in combination with field data. Such an approach still warrants a high cost of inventory. It is therefore useful to explore alternative methods that rely primarily on lidar data by reducing the necessity for field-derived information. The aim of this study was to derive structural attributes of even-aged Eucalyptus grandis forest plantations using lidar data. The attributes are of significance to timber resource assessments and include plot-level tree height attributes, stems per hectare (SPHA), and volume. The surveyed field data included tree counting and measurement of tree height and diameter at breast height for sample plots. Volume was then calculated using standard allometric models. Small-footprint lidar data of the plantations were also acquired coincident with the field inventories. Mean tree height and dominant height were estimated at a range of simulated lidar point densities between 0.25 points/m2–6 points/m2. Various plot-level distributional metrics were extracted from height values of lidar non-ground points and related with field mean and dominant height values using stepwise regression analysis. The results showed that both attributes could be estimated at high accuracies with no significant differences arising from variations in lidar point density. Estimation of SPHA relied on the exploration of semi-variogram range as a mean window size for applying local maxima filtering to the lidar canopy height surface. A comparative approach of window size determination used pre-determined within-row tree spacing, based on planting information. Two secondary objectives were addressed: comparing spatial resolutions of canopy height surfaces interpolated from non-ground height values and comparison of lidar point densities simulated at three levels. Comparison of spatial resolutions of canopy height surfaces were performed at 0.2 m, 0.5 m, and 1 m using a lidar point density of 5 points/m2. The results indicated that 0.2 m is the most appropriate resolution for locating trees and consequently deriving SPHA. Canopy height surfaces of 0.2 m resolution were created at simulated densities of 1 point/m2, 3 points/m2, and 5 points/m2. While all estimates were negatively biased relative to field-observed SPHA, lidar densities of 3 points/m2 and 5 points/m2 returned similar accuracies, which were both superior to 1 point/m2. It was concluded that 3 points/m2 was sufficient to achieve the accuracy level obtained from higher lidar point densities. Plot-level mean height, dominant height, and volume of trees were estimated for trees located using local maxima filtering approaches at the three lidar point densities. Mean height and dominant height were both estimated at high accuracies for all local maxima filtering techniques and lidar point densities. The results were also comparable to the approach that employed regression analysis that related lidar-derived distributional metrics and field measurements. Estimated dominant height and SPHA, as well as age of trees, were used as independent variables in a function to estimate plot-level basal area. The basal area was then used to compute diameter of the tree with mean basal area, referred to as quadratic mean diameter at breast height (QDBH). Mean tree height and QDBH were used as independent variables in a standard equation to calculate mean tree volume, which was then scaled up to the plot-level. All estimates for the local maxima filtering approaches and lidar point densities returned negatively biased volume, when compared to field observations. This was due to the underestimation of SPHA, which was used as a conversion factor in scaling up from tree-level to plot-level. Volume estimates across lidar point densities exhibited similarities. This suggests that low lidar point densities (e.g., 1 point/m2) have potential for accurate volume estimation. It was concluded that multiple forest structural attributes can be assessed using lidar data only. The accuracy of height derivation meets the standards set by field inventories. The underestimation of SPHA may be comparable to other studies that applied different methods. However, improved estimation accuracy is needed in order to apply the approaches to commercial forestry scenarios. The significance of improving SPHA estimation extends to improved volume estimation. In addition, the potential improvement should also take into consideration the density of lidar points, as this will impact on the cost of acquisition. This research has taken a significant step towards determining if lidar data can be used as a stand-alone remote sensing data source for assessment of structural plantation parameters. Not only does such an approach seem viable, but the lower required point densities will help to reduce acquisition costs significantly. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Master of Science - Science / The replacement of indigenous grasslands by commercial forestry plantations on the mountainous eastern escarpment of South Africa, has inevitably led to alteration of nutrient cycling. The overall objective of this study was to determine the nutrient status (soil, litter and foliar nutrient concentrations) of Pinus patula and Eucalyptus grandis plantations and adjacent grasslands. The 2, 13 and 30 year old P. patula plantations and adjacent Driekop grassland were located relatively high up on the escarpment (altitudes ranging between 950–1610 m) and were underlain by dolomite while the 2, 8 and 13 year old E. grandis plantations and adjacent Brooklands grassland were located lower down (altitudes ranging between 900–1100 m) and were underlain by granite. It was assumed that the nutrient status of the soil under the plantations and the adjacent grasslands were similar prior to afforestation. Samples were collected once in winter (August 2002) and once in summer (January 2003). Soil samples were collected to a depth of 20 cm and litter material, to a maximum depth of 20cm. Lower canopy foliar samples were collected within the lowermost metre of the canopies and upper canopy foliar samples were collected within the uppermost metre of the canopies. The data were analysed in the laboratory for various elements and soil physical parameters. The soil pH was lower under the P. patula (pH 4.8-5.2) and E. grandis plantations (pH 4.6-5.3) when compared with the soil under the adjacent grasslands (pH 5.3-5.6). The soil under the P. patula plantations had lower exchangeable base cation concentrations (204-300 mg kg-1) compared with the adjacent Driekop grassland (452-645 mg kg-1), while there were no clear trends when comparing the exchangeable basic cation concentrations in the E. grandis plantations and the adjacent Brooklands grassland. The soil aluminium saturation was similar between the plantations, ranging between 39.7 and 63.3% in the P. patula plantations and 27.4 and 75.6% in the E. grandis plantations. The grasslands had lower soil aluminium saturations, ranging between 17.7 and 35.7% in the Driekop grassland and 17.5 and 39.1% in the Brooklands grassland. Exudates from mycorrhizal associations chelate iv acidic cations, rendering plantations more tolerant of acidic soils. Soil total nitrogen, phosphorus and carbon in the plantations were similar to the adjacent grasslands. Higher nitrogen mineralisation rates in the P. patula plantations corresponded with higher litter and foliar total N concentrations relative to the adjacent Driekop grassland, while there were no clear trends when comparing rates of N mineralisation in the E. grandis plantations and the Brooklands grassland. The concentrations of foliar amino acids and protein were significantly higher (p<0.01) in the plantation foliage, when compared the grassland foliage. The amino acid arginine, occurred in higher proportions relative to other amino acids in plantation foliage, when compared with the relative proportions found in the grasslands. Relative to nitrogen, foliar ratios indicated that phosphorus and potassium were the most limiting nutrients for the plantations, which possibly resulted in N storage in the form of protein and amino acids. The soil and foliage were the more sensitive indicators of the nutrients status than litter. Differences in nutrients concentrations between the lower and upper canopies in both plantations were minor and no major trends were found. Therefore the sub division of lower and upper canopies is not recommended for further foliar nutrient analyses. Age related trends were unclear as a result of higher soil clay contents found in the 13 year old P. patula plantation and the 8 year old E. grandis plantation as well as the various silvicultural practices implemented. In terms of the relative impacts of P. patula and E. grandis plantations, the results of this study imply that the upper 20cm of soil was more impacted under the P. patula plantations than under the E. grandis plantations, in terms of reduced base cation and increased acid cation concentrations. The P. patula plantations are shallow rooters and thus the base cation status of the upper soil horizons may affect the sustainability of the P. patula plantations.

Student Number : 0002920M - MSc dissertation - School of Animal, Plant and Environmental Sciences - Faculty of Science / Sustainability of commercial plantations is dependent on the conservation of soil nutrients, especially on the ancient, leached soils of the tropics. This is particularly important when limited fertilization is practiced. To assess the sustainability of commercial plantations, various sustainability indicators have been suggested. One such indicator is that of biodiversity. The measurement of soil faunal diversity is important as soil fauna play an active role in the cycling of nutrients through the decomposition of plant residues and organic matter. The diversity and different feeding activities of soil fauna influence the rate at which nutrient cycling occurs either through microbial grazing, faecal deposition, the mixing of litter with the mineral soil as well as through the spreading of microbial inoculum. Management practices may, however, negatively influence nutrient availability through losses associated with the burning and removal of slash residues, thus changing the soil faunal community composition. In Eucalyptus grandis plantations in South Africa, the site is usually burned following the harvest and prior to replanting, which may markedly disrupt soil faunal function and nutrient availability. Soil biological processes, nutrient loss and soil faunal composition were the focus of this study. Nutrient losses associated with fire and slash management practices were determined by comparing the quantity of nutrients lost from the slash that was either spread or piled following burning. The in situ nitrogen and phosphorus mineralization rates and soil faunal composition were measured in six randomly located plots, three burned and three unburned, prior to and after a low intensity fire. Results indicate that 78 – 99% of phosphorus, nitrogen and carbon were lost from the slash as a result of burning, with a greater percentage loss occurring from the slash that was spread. It was further shown that within the burned plots the nitrogen availability was enhanced four fold within the first month following the fire. Phosphorus availability was also enhanced ten fold as a result of the burning, however, these effects on nitrogen and phosphorus availability were short lived. Soil faunal diversity was low both prior to and after burning. Ants are the dominant soil fauna. Soil faunal composition was more markedly influenced by season than by fire, with millipedes occurring in all plots in spring, prior to the fire and ants dominating in summer, post fire.

Orientador: Ivan de Godoy Maia / Banca: Luis Eduardo Aranha Camargo / Banca: Adriane Pinto Wasko / Banca: Edson Seizo Mori / Banca: Esteban Roberto González / Resumo: Os cassetes de expressão empregados atualmente para a produção de plantas geneticamente modificadas são baseados, na sua maioria, em promotores constitutivos que determinam a expressão generalizada do transgene na planta, o que muitas vezes não é necessário nem desejado. A alternativa mais viável para substituição de tais promotores é investir na identificação e caracterização de promotores órgão/tecidoespecíficos ou estímulo-dependentes nas espécies de interesse. O presente projeto teve como objetivos identificar e caracterizar genes com padrão de expressão órgãoespecífico em eucalipto e, a partir dessas informações, isolar e caracterizar as seqüências promotoras adjacentes. Predições in silico no banco de dados do projeto de seqüências expressas do eucalipto (FORESTs), e informações disponíveis na literatura a respeito de genes com padrão de expressão órgão-específico, foram utilizadas na seleção. Assim, 59 genes preditos como possuindo expressão exclusiva em dado órgão foram selecionados para a validação via RT-PCR. Dos candidatos validados, 2 eram de raiz, 1 de folha, 1 de botão floral e 1 de botão floral e fruto, concomitantemente. Três candidatos ubíquos também foram selecionados. Dois desses candidatos tiveram seus perfis de expressão em diferentes órgãos de eucalipto avaliados por PCR quantitativa, indicando que o primeiro é preferencialmente expresso em folhas, e o segundo expresso especificamente em raiz. A partir de estratégias de "genome walking" foram isolados diferentes promotores putativos, sendo que o promotor do candidato com expressão preferencial em folha (1.2 kb) foi selecionado para a caracterização funcional em plantas usando o gene uidA, que codifica a -glucuronidase (GUS), como repórter. Em plântulas transgênicas de tabaco da geração T1, a expressão de GUS foi detectada majoritariamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Many plant genetic engineering applications require spatial expression of transgenes, which in turn depends upon the availability of tissue- and organ-specific promoters. In the present work, the identification of genes with organ-specific expression in Eucalyptus grandis was performed aiming the subsequent isolation and characterization of contiguous promoter sequences. Candidates genes were selected by in silico predictions in the Eucalyptus EST project (FORESTs) database or by searching the available literature for genes with specific expression patterns. Fifty-nine genes with predicted organ-specific expression were selected for further RT-PCR validation. Among the validated candidate genes, 2 were root-specific, 1 was leaf-specific, 1 was flower-bud-specific and 1 was flower-bud and fruit specific. Three genes ubiquitously expressed were also selected. The relative expression levels of two of them (one leafspecific and one root-specific) over a broad range of eucalyptus organ/tissues were determined using real time PCR. The 5' putative regulatory sequences of the validated genes were isolated using genome walking strategies, and the activity of the leafspecific promoter (1.2 kb) was further investigated in transgenic tobacco plants using a GUS reporter system. In transgenic seedlings from the T1 generation, GUS expression driven by this promoter was detected preferentially in cotyledons. In adult plants, however, GUS expression was detected in both stem and root, but in lower intensity if compared to the expression observed in leaves. This promoter was also able to modulate the transient expression of GUS in seedlings of Eucalyptus grandis. / Doutor

Orientador: Ivan de Godoy Maia / Banca: Celso Luis Marino / Banca: Marcio José da Silva / Resumo: A identificação de promotores com expressão tecido-específica é uma alternativa viável para substituição dos promotores com expressão ubíqua geralmente utilizados em transgenia. Nesse contexto, o presente trabalho teve como objetivos caracterizar funcionalmente o promotor de um gene de eucalipto que codifica um transportador de potássio com expressão específica em raiz bem como isolar e caracterizar a região promotora de um gene de eucalipto selecionado como apresentando expressão específica em folha. Plantas transgênicas de tabaco (Nicotiana tabacum SR1) contendo um cassete de expressão composto pelo promotor de raiz fusionado ao gene repórter GUS (que codifica a β-glucoronidase) foram usadas em ensaios histoquímicos e histológicos para investigar a especificidade da expressão determinada pelo promotor em estudo. Os resultados evidenciaram que o promotor investigado dirige a expressão do gene repórter em tecido vascular de folhas e raízes. A expressão em feixes vasculares de folhas e raízes foi confirmada em cortes histológicos. Visando avaliar a resposta deste promotor a baixas concentrações de potássio, duas linhagens da geração T2 foram submetidas à deficiência de potássio, e a expressão relativa do gene repórter GUS foi determinada por PCR em tempo real. Nas linhagens submetidas a estresse de potássio observou-se um aumento da expressão relativa do gene repórter GUS em função da privação do elemento. Em paralelo, um gene selecionado in silico como apresentando expressão em folha de eucalipto teve seu perfil de expressão validado por RT-PCR. A construção de um cassete de expressão contendo o referido promotor fusionado ao gene repórter GUS foi empreendida visando futuras validações funcionais / Abstract: The identification of tissue-specific promoters is of great value to substitute the ubiquitous promoters generally used in transgenic production. In this context, the present study aimed to functionally characterize the promoter of a Eucalyptus grandis gene encoding a potassium transporter showing root specific expression, and to isolate and functionally characterize the promoter region of an E. grandis gene selected as showing specific expression in leaf. Transgenic tobacco plants (Nicotiniana tabacum SR1) harboring a promoter:GUS fusion were used to investigate the expression specificity of the selected root promoter. The results showed that the investigated promoter drives a reporter gene expression in vascular tissues of leaves and roots. The expression in vascular bundles of leaves and roots was confirmed in histological crosssections. To evaluate the promoter responsiveness to low potassium concentrations, two transgenic lines (T2) were submitted to potassium starvation and the relative expression of the GUS reporter gene was determined by real time PCR. An increase in the relative expression of GUS in response to potassium starvation was observed. In parallel, the expression pattern of a gene showing leaf specific expression in Eucalyptus grandis was validated by RT-PCR. The construction of an expression cassette containing this promoter fused to the GUS reporter gene was performed aiming future functional characterization / Mestre

It is believed that the process-based model 3-PG (Physiological Principles Predicting Growth; Landsberg and Waring, 1997) can potentially play a useful role within South African forestry, both as an operational and a strategic tool. Strategic applications may include the prediction of potential productivity on a site-by-site basis; broadscale productivity estimates based on remote sensing and the spatial application of 3-PG; identification of production constraints; and estimation of carbon fluxes to help address sustainability issues. Operationally, 3-PG could complement empiricallybased models or be used in conjunction with them as a hybridised product. The challenges of this study were therefore to see whether it is possible to adapt 3-PG to predict the growth and yield of E. grandis under South African conditions, test that model predictions are consistent with observed growth data and are biologically reasonable, and to assess the practicality of using 3-PG as either a strategic or operational tool. The main emphasis of this study was to understand the internal logic of 3-PG and how physiological processes are represented, and to develop methods to objectively parameterise and initialise the model. Thereafter a detailed model validation study was performed, ending off with selected potential applications of 3-PG within the South African context. The sensitivity of predicted stand volume (SV) and leaf area index (LAI) to the values of the species-specific parameters in 3-PG was examined. These analyses enabled the development of three distinct parameter sensitivity classes: insensitive parameters (i.e. those that can be varied widely without affecting the outputs studied), sensitive parameters (i.e. those whose value strongly affects the outputs, and non-linear parameters (i.e. those for which the outputs depend in a non-linear manner on the parameter value). Minimum data requirements for the parameterisation and initialisation of 3-PG are considered in detail. Conventional methods used for the parameterisation of models, specifically 3-PG, are reflected upon. An automated parameter estimation technique was examined and used for the estimation of selected parameters. Species-specific parameters were categorised according to data source estimation and sensitivity classes. Parameters describing allometric and age-dependent relationships were assigned values using observed data from biomass harvests. Critical parameters that could not be directly assigned using observed data were the ratio of foliage to stem allocation (i.e. P2 and p2o), allocation of net primary production (NPP) to roots (TJRX and T]Rn), optimum temperature for growth (7^,) and maximum canopy quantum efficiency (acx)- These were estimated using Parameter ESTimation, by fitting model output to corresponding observed growth data. As well as species-specific parameter values, mandatory inputs required by 3-PG include weather data, site-specific factors such as site fertility (FR) and physical properties of the soils, and stand initialisation data. Objective techniques to determine these site-specific factors and the initial values for the biomass pools were proposed. Most of these data are readily available for sites where experimental trials exist, or where monitoring networks are in place. However, this is the exception rather than the rule, so alternative data and information sources are required. These, together with the need for accurate weather inputs (especially monthly rainfall) and physical properties (especially soil texture, maximum available soil water and FR) of the sites being modelled were explored. 3-PG was validated using four simple tests by comparing predicted versus observed SV. Results showed that 3-PG predictions are relatively consistent with observed stand data. Analyses performed using time-series data showed model predictions accurately tracked observed growth in response to erratic and fluctuating weather conditions. Results from the initial model validation showed production on high and low productivity sites was under- and over-predicted, respectively. Further results presented here show a similar, but less marked trend (i.e. over- and under-predictions are not as extreme), and individual biases are less than those from predictions made using another locally developed parameter set. The application of 3-PG showed that the model is able to make estimates of tree growth that are consistent with those used within the forestry site classification. This showed the considerable potential 3-PG has for strategic planning by the forest industry (i.e. projected wood supplies etc) and in research planning (refining existing site classifications). The model could be useful in predicting growth in various areas where E. grandis is not grown, assisting in future decision making. 3-PG was able to identify growth constraints on a site-by-site basis and distinguish among them, and was able to identify environmental and site limitations to plantation growth, and how they vary in space and time. These results together with predictions of site productivity demonstrate the potential for 3-PG to be used to improve existing forest site classifications. The model comparison study between empirically-based models and 3-PG showed that although the empirical models made accurate predictions of volume under static climatic conditions, under fluctuating weather conditions empirical estimates of volume were less accurate than those made with 3-PG. 3-PG can therefore be used operationally with minimum input data to predict growth using enumeration data. This is useful in saving time and cutting costs. The use of process-based models (PBMs) in general, and 3-PG in particular, needs to be "championed'' to the South African forest industry. This is necessary for two reasons. Firstly, the model and the manner with which it describes physiological processes of growth need to be explained in layman's terms. This will demonstrate how and why a process-based model can work better in a fluctuating environment than empirically based models. Secondly the comparison between 3-PG and the local empirical models needs to be presented as an example of how 3-PG can be applied on an operational basis. It is accepted that much convincing is still required. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2005.

In South Africa, Eucalyptus breeding programmes are aimed at the selection of fastgrowing varieties, with appropriate wood characteristics and/or resistance to pests and diseases. However, the slow growth rate, long generation time and heterozygosity of trees make this a difficult task. Such problems may be overcome by the adoption of a biotechnological approach for plant propagation and modification. Towards this end, the aims of this investigation were to establish protocols for the micropropagation of Eucalyptus grandis and for the Agrobacterium-mediated transformation and subsequent plant regeneration of this important species. The usefulness of transformed cells and/or tissues is dependent upon the availability of methods for their regeneration into plants. Consequently, methods for plant regeneration via indirect organogenesis from leaf discs and cell suspension cultures were investigated. Organogenic calli were produced from leaf explants on MS medium with 16 mg.1-1 &l:em•calcltrate, 20 g.I-I sucrose, 1mg.I-I NAA and 0.05 mg.1--<1 BA. Shoots were induced on MS medium containing 1 mg.1-1 ZEA and 0.2 mg.1-1 IAA, and subsequently rooted on medium containing MS nutrients supplemented with 1 mg.1- 1 IAA. Cell suspension cultures were established but not regenerated via indirect organogenesis. Additionally, various media were investigated in order to obtain somatic embryos from cell suspension cultures. The MS media supplemented with 30 g.1-1 sucrose, 12 mg.1- 1 ABA and/or 40 g.1-1 PEG were found to be most suitable, resulting in the production of embryoids; germination results are not available at this stage. In order to establish methods for the transformation of both leaf discs and cell suspension cultures of Eucalyptus, a triparental mating was performed between Escherichia coli pnT119 (donor), A. tumefaciens LBA4404 (recipient), and E. coli HBI0l::pRK2013 (helper), resulting in the transconjugant LBA4404 (pnT119); insertion of the pJIT119 plasmid was demonstrated using agarose gel electrophoresis. The transconjugant CS8C1 (pMP90) (pJIT119) was also used. Protocols for the transformation of both leaf discs and cell suspension cultures were established, and resulted in the production of putatively transformed calli which were GUS positive and with stood selection on kanamycin (50 Ilg.mr1) and/or sulfadiazine (50 Ilg.mr1). Also, Southern blotting analysis indicated that the gene transfer process was successful. Due to difficulties in the regeneration of plants from transformed calli transgenic plants were not obtained. Future research strategies and applications of the developed protocols to Eucalyptus breeding programmes are discussed. / Thesis (M.Sc.)-University of Natal, 1994.

South Africa depends largely on afforestation programs for its timber supplies due to the great demands for fiber and wood products. This has brought discomfort to other water users who have advocated that the effects of afforestation on water resources are detrimental to the country as a whole since South Africa is known as a water scarce country. This study has undertaken to integrate a process-based model and remote sensing data to estimate water use and productivity of Eucalyptus grandis in the Zululand areas of South Africa. The remote sensing techniques and recently developed "process based model" that is 3PG-S were used to estimate water use and productivity of Eucalyptus grandis, an economically important plantation species grown in the summer rainfall areas of South Africa. The study utilized monthly Landsat Thematic Mapper datasets and climatic data as inputs into the 3PG-S model, determined the Leaf Area Index (LAI) and Specific Leaf Area (SLA) through direct (destructive sampling) and indirect measurements (LiCor- 2000) and assessed the relationships between various vegetation indices (VI's) using correlation and regression analyses. The results suggest that all the indices, except the ratio VI, correlated significantly with LiCor-determined and destructively measured LAI values with both normalized difference vegetation index (NDVI) and Ratio Vegetation Index (RVI) (r=0.86, p / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritburg, 2007.

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.