Thesis (Ph.D.)--Murdoch University, 1999. / Thesis submitted to the Division of Science. Bibliography: leaves 190-217.

This thesis examines a broad spectrum of physiological and silvicultural features of the highly valued woody angiosperm hemi-parasite Santalurn album L. (Indian sandalwood) in relation to its culture in plantations in northern Western Australia. Topics covered include allometry of host and Santalum when grown as single plant pairings in both field and pot culture, nutritional interactions between Santalum and beneficial and non-beneficial hosts, deleterious influences of parasitism on plantation productivity and heartwood induction in young trees. In Western Australia sandalwood is grown in the nursery for 8 months before establishment in the field and during this time a pot host is introduced. Survival of Santalurn after field establishment and its subsequent growth were significantly affected by the time of introduction of the pot host, Alternanthera nana. Increasing the period of the Santalum : Alternanthera association in the nursery to 109 days prior to field establishment markedly increased early growth of Salztalum plantations. Introduction at 134 days prior to field establishment was detrimental to the parasite as the Alternanthera was too vigorous for the small Santalum seedlings. Santalurn plants had a lower root : shoot ratio lower when cultured with Alternanthera in the nursery prior to field establishment compared with seedlings grown without Alternanthera. Alterrzantlzera survival in the field was high when it had been grown with Santalum for 12 weeks or more in the nursery prior to field establishment. After 1 1 weeks in the field a strong negative linear relationship was shown between Santalunz root : shoot ratio and Alternarzthera dry weight, and a positive linear relationship between Salztalum DW and Alternanthera DW. In Western Australia Santalu~n is established in the field with an intermediate host which nourishes the parasite for 3-5 years before Santalum becomes dependent on its long-term host and the intermediate host dies. The relationship between Santalum and several species tested as intermediate hosts was examined by pairing Santalum seedlings with intermediate host seedlings in 25 litre pots over a 10 month period. Growth of Santalum in pot culture with three N2-fixing woody intermediate hosts (Sesbania forrnosa, Acacia traclzycarpa and A. ampliceps), the woody non N2-fixing Eucalyptus camaldulensis or without a host varied considerably between host treatments. Santalum growth was greater and root : shoot ratio lower for seedlings grown with N2-fixing hosts compared with seedlings grown with E. carnaldulensis or with no host. The root : shoot ratio of unattached Santalum increased exponentially over time, whereas for all other treatments it remained relatively constant. An assessment of the value of the hosts, termed host use efficiency, was computed as Santalum shoot DW / host shoot DW. The host use efficiency of A. trachycalpa was greater than that of the other hosts. The xylem sap of hosts and Sarztalum, and ethanolic extracts of endophytic tissue of haustoria of Santalzkm were analysed for amino acids, organic acids and sugars to determine which solutes were available in the host and which were extracted by the Santalum haustoria from different hosts. There were similarities between Santalum and legume hosts in concentration and composition of xylem sap amino acids, and in the amino acid spectra of the corresponding Santalum endophytic tissue, whereas there were low N levels in xylem sap of E. camaldulensis and dissimilarities between its amino acid composition and that of Santalum. This indicated substantial direct intake of xylem N by Santalum from legume hosts but little N from the xylem sap of E. canzaldulensis. There were high concentrations of asparagine, glutamate, aspartate and y-amino glutamate in the xylem sap of the legume hosts, while in the non-legume the most common amino acids were glutamate, aspartate, glutamine and arginine. Proline, the predominant amino acid in the xylem sap of Santalum acurninatum growing in natural vegetation (Tennakoon et al. 1997) was not detected or present in very low concentrations in Santalurn album under these conditions. in the non-legume. Xylem sap of hosts contained variable amounts of sugars (sucrose, glucose and fructose) and organic acids (fumaric, citric and malic acid), whereas that of the parasitic Santalum was dominated by fructose and malic acid. Dissimilarities in the proportional amounts of xylem-borne sugars and organic acids were particularly evident for the E. camaldulensis : Santalum partnership. Diurnal profiles of photosynthesis and transpiration of Santalum were closely similar to those for corresponding hosts, whereas the midday leaf water potential of Santalum was consistently more negative than that of corresponding hosts. Net photosynthesis and water use efficiency was lower, but transpiration rates were similar to that of corresponding hosts. Nitrogen concentrations of foliage of Santalum were higher than their hosts, and higher when on legume hosts than on E. camaldulensis, or without a host. Nitrogen concentrations of Santalum foliage was strongly correlated with net photosynthesis and water use efficiency of Santalum. 813C values of shoot dry matter of Santalum were poorly correlated with instantaneous water use efficiency of Santalum. Tissue water relations of Santalum were similar to that of water-stress tolerant species. S. formosa proved the best host followed by Acacia ampliceps and A. traclzycarpa based on dry matter gains of Santalum. Estimates of heterotrophic gain of C of Santalum when grown in association with the legume hosts over a nine week period indicate 57.9% of C was derived from A. ampliceps, 45.5% from A. trachycarpa and 34.6% fiom S. fomosa. Abundance of haustorial attachments on roots of hosts was poorly correlated to Santalum shoot DW. Root nodules of legume hosts were parasitised by a small proportion of Santalum haustoria. Sodium and phosphorus concentrations of foliage of Santalum were generally higher than that of corresponding hosts. Net gains of calcium, potassium, phosphorus and sodium in Santalum was greatest when grown in association with hosts richest in the corresponding element. Net losses or only small gains of calcium, potassium, phosphorus and sodium were recorded when Santalum was grown with E. camaldulensis or without a host suggesting that Santalum has limited ability for uptake of those minerals through its own root system. To understand the effect of hosts on the productivity of a Santalum plantation a young plantation of Santalum with three host species Cathormion umbellatum, Sesbania formosa and Acacia anuera was selected to study the relationship between host quality and distance of hosts from Santalunz on Santalum health. The selected plantation showed marked decline in health and vigour of both Santalum and hosts between years 3 and 5. Parameters of the host plants were assessed to select the best predictor of Santalunz crown health. The height and diameter growth increment of Santalum between years 3 and 5 was strongly correlated to Santalum crown health. Santaluin crown health and growth increased as host quality increased, and the distance of host fiom Santalum decreased. An index, which combined host quality and the distance of the host from that of Santalum, was a better predictor of Santalum crown health than host distance or quality alone. The age at which heartwood is initiated in Santalum album under plantation conditions in Western Australia in unknown, but in natural stands in India it occurs between 10-13 years of age (Rai 1990). A field experiment was conducted to determine the efficacy of stem injections of paraquat andlor ethrel in initiating heartwood formation in five year old Santalum trees in a plantation. Trees injected with paraquat alone had a significantly greater extension of induced heartwood, both radially and vertically, than those trees injected with ethrel alone or distilled water. Eight months after treatment with paraquat or ethrel or a combination of these chemicals induced heartwood was formed, which had high lipid, and low starch and polysaccharide concentrations compared to the sapwood. Induced heartwood from both chemical treatments and their combinations contained total volatile oil and santalol oil (alpha and beta santalol) concentrations that were equal to or greater than that of naturally formed heartwood and greater than that of sapwood. Moisture content, and concentrations of K and Mg, and in some treatments Ca of induced heartwood were significantly lower than that of sapwood. The thesis concludes with a synthesis of the findings and suggestions for future research, with special reference to mid-rotation aspects of Santaltrm plantation silviculture.

<p>  </p> <p><em>Santalum</em> spp., known globally as sandalwood, are highly sought after for their aromatic oil-rich heartwood and have been exploited throughout their range. Six of the 19 <em>Santalum</em> species are endemic to the main Hawaiian Islands, where they are known locally as ʻiliahi. Excessive harvesting led to the extirpation of Hawaiian sandalwood from 90% of its historic range by 1840. There is limited peer-reviewed literature concerning the propagation of Hawaiian sandalwood, and methods developed for other non-Hawaiian species cannot be directly adopted due to differences in sandalwood species physiology, available host species, and growing environment. This, combined with increased interest in growing Hawaiian sandalwood, prompts the need for the development of propagation protocols based on empirical research. The primary knowledge gaps in propagation include best practices for producing high-quality seedlings in the nursery (e.g., fertilizers and hosts) and silvicultural practices for maximizing outplanting survival and growth. The Hawaii Island endemic <em>Santalum paniculatum</em> has the largest remnant population and distribution and high commercial value, making it an ideal species to focus our study on. We conducted two experiments to evaluate the response of <em>S. paniculatum</em> seedlings to propagation methods employed with Australian and Indian sandalwood, although with species of hosts native to Hawaii. The first experiment was a nursery growth trial that evaluated the quality of <em>S. paniculatum </em>seedlings in response to nutrient availability (controlled-release fertilization, control), chelated iron fertilizer (applied, control), and species of pot host (<em>Acacia koa</em>, <em>Dodonaea viscosa</em>, control). The quality of seedlings was determined by measurements of height, root collar diameter, dry mass, root shoot ratio, chlorophyll index, and nutrient status (N & Fe concentration). Nutrient availability had the greatest impact on seedling quality and increased height, root collar diameter, dry mass, chlorophyll index, and nutrient status. Chelated iron fertilizer effectively improved seedling quality (height, collar, dry mass, chlorophyll content, and Fe concentration) in a nutrient-limiting environment, although a nutrient-rich environment diminished its effect with sufficient iron levels. The host species treatment had the least influence on seedling quality and only influenced haustoria formation by causing more haustoria in <em>A. koa</em>-paired compared to <em>D. viscosa</em>-paired and control seedlings. Although the pot host had the lowest effect on seedling quality during nursery propagation, it provided benefits in the field planting phase of the project.</p> <p>The second experiment of the project assessed the survival and performance of field-planted <em>S. paniculatum</em> in response to (1) nursery fertilization, (2) an <em>A. koa</em> pot host, and (3) an <em>A. koa </em>field host. Nursery fertilization had the greatest effect on performance and enhanced survival, height, height growth, collar, collar growth. The survival rate of unfertilized seedlings was 43.3% (± 5.9) compared to 86.9% (± 4.2) for fertilized seedlings. The pot host improved height, height growth, collar, collar growth of seedlings, but it did not influence survival. The intermediate field host significantly improved survival from 52.7% (± 7.8) to 78.0% (± 5.6) and only affected the height measurements and not the collar. There was a significant interaction between the field host and nursery fertilizer treatment associated with the fertilized seedlings planted with field hosts having lower water potential than the fertilized seedling planted without a field host. Our results demonstrated that supplemental nursery nutrition, pot hosts, and intermediate <em>A. koa </em>field hosts benefited <em>S. paniculatum</em> regeneration establishment in different ways. Furthermore, the effect of the hosting treatments may become more pronounced in the field over time as more haustoria connections are formed. This research project provides essential baseline information that helps to enhance the current methodology and inform future decision-making concerning the propagation of <em>S. paniculatum </em>and other Hawaiian <em>Santalum</em> species. </p>

Calcium ion plays a pivotal role as second messenger during signal/response coupling in plant cells (Trewavas, 1999). Elevations of cytosolic Ca2+ occur in plants as a consequence of abiotic and biotic stresses, environmental and hormonal stimuli. However, the molecular mechanism by which changes in cytosolic calcium are sensed and transduced in the plant cell has not been completely elucidated. The detection of Ca2+-binding proteins, especially Ca2+-dependent protein kinases (CDPKs) in plants led to drawing analogy with animal systems wherein the Ca2+-message is perceived and transduced by proteins that bind Ca2+. CDPKs are stimulated by the direct binding of Ca2+ to their endogenous calmodulin (CaM) -like domain (Harper et al, 1991). CDPKs exist as multiple isoforms in a single species, and show tissue-specific and developmentally regulated expression. Furthermore, the diversity among different CDPK isoforms with respect to Ca2+-binding properties, activation, substrate specificity, regulatory mechanisms and other kinetic properties suggest their specialization in the regulation of distinct signaling pathways. These observations therefore have led to the speculation that most of the Ca2+-mediated signal transduction in plants occurs via the mediation of CDPKs (Harmon et al, 2000). Over the last 15 years there has been a dramatic unfolding of information on Ca2+-mediated signaling in plants. Nevertheless, little is known about the environmental/hormonal signals and the signaling events that regulate early plant developmental processes such as embryogenesis, seed development and germination. The present investigation was initiated with the objectives 1) to determine the role of Ca2+ during embryogenesis, 2) to examine the involvement of a CDPK during early developmental processes in sandalwood plant (Santalum album L.) and 3) to purify and biochemically characterize this CDPK. The study initially investigated the possible involvement of calcium-mediated signaling in the induction/regulation of somatic embryogenesis from proembryogenic cells of sandalwood. 45 Ca + uptake studies and fura-2 fluorescence ratio photometry were used to measure changes in [Ca2+]cyt of proembryogenic cells in response to culture conditions conducive for embryo development. Sandalwood proembryogenic cell masses (PEMs) were obtained in the callus proliferation medium that contains the auxin 2,4-D. Subculture of PEMs into the embryo differentiation medium which lacks 2,4-D and has higher osmoticum resulted in a 4-fold higher 45Ca2+ incorporation into the symplast. Fura-2 based ratiometric analysis also showed a 10-16- fold increase in the [Ca2+]cyt of PEMs under identical culture conditions, increasing from a resting concentration of 30-50 nM to 650-800 nM. Chelation of exogenous Ca2+ with EGTA arrested such an elevation in [Ca2+]cyt. Exogenous Ca2+ when chelated or deprived also arrested embryo development and inhibited the accumulation of a Ca2+-dependent protein kinase (swCDPK) in embryogenic cultures. However, such culture conditions did not cause cell death as the PEMs continued to proliferate to form larger cell clumps. Culture treatment with W7 reduced embryogenic frequency by 85%, indicating that blockage of Ca2+-mediated signaling pathway(s) involving swCDPK and/or CaM caused inhibition of embryogenesis. These observations suggest a second messenger role for exogenous Ca2+ and the existence of Ca2+-mediated signaling pathway(s) during sandalwood somatic embryogenesis. The detection of a 55 kD protein showing cross reactivity with polyclonal antisoybean CDPK and the detection of Ca2+-dependent protein kinase activity in protein extracts from somatic embryos, prompted investigation on the spatio-temporal accumulation and activity of a CDPK in different developmental stages of sandalwood. Western blot analysis and protein kinase assays identified a Ca2+-dependent protein kinase (swCDPK) of 55 kD in soluble protein extracts of different developmental stages of sandalwood somatic embryos. However, swCDPK was not detected in plantlets regenerated from somatic embryos. swCDPK exhibited differential expression and activity in the developmental stages of sandalwood. Zygotic embryos, endosperm and seedlings showed high accumulation of swCDPK. However, the enzyme was not detected in the soluble proteins of shoots and flowers of sandalwood tree. swCDPK exhibited a temporal pattern of expression in endosperm, showing high accumulation and activity in mature fruit and germinating stages, the enzyme being localized strongly in the storage bodies of the endosperm cells. Interestingly, these storage bodies were thereafter identified as oil bodies, suggesting that a Ca2+-mediated regulation of oil hydrolysis and/or mobilization might be operative during seed germination. swCDPK in the zygotic embryo was found to be inactive during seed dormancy and early stages of germination, indicating a possible post-translational hibition/inactivation of the enzyme during these stages. The temporal expression of swCDPK during somatic/zygotic embryogenesis, seed maturation and germination thus suggests involvement of the enzyme in these early developmental processes. In view of the diversity exhibited by members of the CDPK family, characterization of swCDPK, the early development specific CDPK from sandalwood was undertaken. Purification of swCDPK was achieved by chromatography on DEAE-cellulose, hydroxyapatite and Blue-Sepharose. The purified enzyme resolved into a single band on 10 % polyacrylamide gels, both under denaturing and non-denaturing conditions. swCDPK was strictly dependent on Ca2+, K0.5 (apparent binding constant) for Ca2+-activation of substrate phosphorylation activity being 0.7 μM and for autophosphorylation activity —50 nM. Ca2+-dependence for activation, CaM-independence, inhibition by CaM-antagonist (IC50 for W7 = 6 μM, for W5 = 46 μM) and cross-reaction with polyclonal antibodies directed against the CaM-like domain of soybean CDPK, confirmed the presence of an endogenous CaM-like domain in the purified enzyme. Kinetic studies revealed a Km value of 13 mg/mL for histone III-S and a Vmax of 0.1 nmolmin-1rng-1. The enzyme exhibited high specificity for ATP with a Km value of 10 nM. Titration with Ca2+ resulted in enhancement of the intrinsic emission fluorescence of swCDPK and a shift in the λmax emission from tryptophan residues. A reduction in the efficiency of non-radiative energy transfer from tyrosine to tryptophan residues was also observed. These are taken as evidence for the occurrence of Ca2+-induced conformational change in swCDPK. The emission spectral properties of swCDPK in conjunction with Ca2+ levels required for autophosphorylation and substrate phosphorylation help elucidate the possible mode of Ca2+ activation of this enzyme.

La présente étude porte sur l’analyse fondamentale et expérimentale des procédés d’extraction des volatils. Les méthodes conventionnelles d’hydrodistillation et d’entrainement à la vapeur dans l’extraction des composés volatils (huiles essentielles…), impliquent une cinétique ralentie voire bloquée par « le processus paradoxal ». Ce dernier est dû au couplage de la génération interne de vapeur impliquant deux transferts couplés de vapeur (par diffusion) et de chaleur (par conduction). Ces deux transferts sont alors dirigés de la surface d’échange vers l’intérieur. Seul un processus de transfert partiel de la vapeur par front progressif assure l’évolution du processus. La cinétique est ainsi inévitablement lente, générant un niveau élevé de consommation d’énergie et induisant une dégradation potentielle de la matière extraite. Pour faire face à cette situation, une modification du type de transfert est essentielle. Nous avons opté pour un transfert de type Darcy au lieu du transfert de Fick, soit un gradient de pression totale au lieu du gradient de pression partielle comme « driving force ». Nous avons ainsi choisi la Détente Instantanée Contrôlée DIC comme procédé générant, suite à la détente vers le vide, une autovaporisation instantanée couplée à un grand gradient de pression totale de l’intérieur du matériau vers le milieu environnant sous vide. La détente instantanée contrôlée (DIC) a été étudiée, selon la méthodologie des plans d’expériences dans le but d’évaluer l’impact des variables opératoires sur l’extraction et de définir les paramètres optimums. Le nombre de cycles et le temps de traitement ont été pris comme paramètres indépendants. Ces paramètres sont appropriés pour représenter les aspects mécaniques et thermiques du processus DIC. L’extraction directe de l’huile essentielle du bois de santal par DIC-MP, gérée essentiellement par le nombre de cycles, indépendamment du temps de traitement, est une autovaporisation des composés volatils. Cette nouvelle méthode a ensuite été comparée tout au long de sa mise en application à l’hydrodistillation. Alors que la méthode traditionnelle d’extraction de l’huile essentielle du bois de santal requiert 140 h, l’optimisation de la DIC nous a permis de réaliser l’extraction en seulement 6 minutes, avec des rendements pratiquement similaires. Dans un dernier temps, une approche écologique et une compréhension du phénomène mis en jeu pour l’extraction des huiles essentielles par DIC ont été abordées. La rapidité de la procédure, la réduction du coût énergétique et le bon impact environnemental font de la DIC une technique industrielle particulièrement innovante et hautement attrayante. / This study focuses on fundamental analysis and experimental work carried out on extraction via conventional and innovative processes. The extraction of volatile compounds (essential oils ...) through hydrodistillation or steam distillation as conventional methods is slowed or even blocked because of "the paradoxical process". The latter is due to the coupling of vapor and heat transfers, both directed from the exchange surface towards product core. Only partial vapor transfer by progressive forehead ensures the evolution of the process. The kinetics is so inevitably slow, implying high energy consumption and inducing a high degradation of both extracted and residual material. To face this situation, changing the nature of the transfer is essential. We considered the total pressure gradient instead of the vapor pressure gradient as "driving force". Instant Controlled Pressure Drop DIC was used. Instant releasing of high pressure/high pressure towards a vacuum leads to instant autovaporisation, implying a Darcy-type transfer of vapor within the material to the surrounding medium vacuum. Instant Controlled Pressure Drop (DIC) was investigated, according to the Response surface methodology RSM in order to evaluate the impact of process variables on the extraction and to define the optimal settings. The number of treatment cycles and treatment time were taken as independent parameters. These parameters were appropriated to represent both mechanical and thermal impacts of DIC process. Direct extraction of the essential oil of sandalwood DIC-MP, managed primarily by the number of cycles, regardless of treatment time, is an autovaporization of volatile compounds. This new method was then compared throughout its application to steam distillation. While traditional extraction method requires 140 hours, optimized DIC allowed performing essential oil extraction of sandalwood in only 6 minutes, with substantially similar yields. Finally, environment approach and understanding of the phenomena involved in DIC extraction of essential oils were addressed. By dramatically reducing the treatment time and the energy consumption, DIC is a particularly innovative, exceptionally effective and highly attractive environment-friendly essential oil extraction technique.

[Truncated abstract] An investigation into the causes of heartwood and essential oil content of Australian plantation sandalwood, Santalum album was undertaken. Genetic diversity of 233 S. album, five S. austrocaledonicum and fifteen S. macgregorii trees growing in the Forest Products Commission arboretum, Kununurra WA, was assessed using nuclear and chloroplast RFLPs. Santalum spicatum was chosen as an out-group. Nuclear genetic diversity of the S. album collection was very low, with observed and expected heterozygosity levels of 0.047. This was lower than the results previously reported in the literature for trees in India, however a different technique was used. Based on allelic patterns, the collection was able to be categorised into 19 genotypes; each representing some shared genetic origin. Some groups were highly redundant with 56 trees being represented, while others were populated by just one tree. The essential oil yield and heartwood contents of trees from these genetic groups were compared. Yields were highly variable both within and between groups of trees which share a common genetic history, suggesting a significant environmental component was contributing to the observed phenotype, despite identical soil and climatic conditions. Ancestral lineages were tested using chloroplast RFLPs, although a lack of shared mutations between species made this difficult. Only one S. album tree originating from Timor was resolved using nuclear RFLPs, with the other trees being grouped with material sourced from India. There was no resolution of Indian S. album from Timorese using chloroplast RFLPs, however one S. album tree grown from Indian seed possessed a single unique mutation. The low genetic diversity of the Australian S. album collection is likely to be a combination of incomplete seed sourcing and highly restricted gene flow during the evolution of the species. Combined with information gathered on the phylogeny of the genus by other researchers, S. album is postulated to have originated from an over-sea dispersal out of northern Australia or Papua New Guinea 3 to 5 million years ago. Essential oil yield and composition was assessed for 100 S. album trees growing in the collection, ranging in age from 8 to 17 years. Oil content of heartwood ranged from 30 mg g-1 to 60 mg g-1, and the transition zone 36 mg g-1 to 90 mg g-1. Sapwood contained almost no sesquiterpene oils. Despite the highly variable total oil yields, the chemical profile of the oil did not vary, suggesting there was limited genetic diversity within this region of the genome. Strong, positive correlations existed between v sesquiterpenoids in the essential oil of S. album. ... These represent the first TPS genes to be isolated from sandalwood and will enable further elucidation of oil biosynthesis genes. This thesis compiles a three-pronged approach to understanding the underlying causes of oil yield variation in S. album. As a species for which so little is known, the research presented here provides a major leap forward for tree improvement, breeding and silviculture. Hence the best of Santalum album research is presented.

The objective of this thesis is to develop an ecological economic framework for the assessment and establishment of protected areas (PAs) that are aimed at conserving forests and biodiversity. The framework is intended to be both rigorous and relevant to the decision-making process. Constructivism is adopted as the paradigm guiding the research process of the thesis, after firstly examining also positivist philosophy and ???post-normal??? scientific methodology. The tenets of both ecological and environmental economics are then discussed. An expanded model of human behaviour, which includes facets derived from institutional economics and socioeconomics as well as aspects of neoclassical economics, is outlined. The framework is further developed by considering, from a contractarian view point, the implications of intergenerational equity for biodiversity conservation policies. The issues of intragenerational distribution and allocation are then considered. In this regard, cost-benefit analysis (CBA), as applied to the valuation of forests, PAs, and biodiversity, is critically reviewed. A participatory approach to decision-making, which may also include CBA, is then proposed. The resulting ecological economic framework may be thus summarised: (a) ecosystem use patterns should be chosen on the basis of their sustainability, distributional, and efficiency aspects; (b) systems of PAs should be established in order to achieve minimal intergenerational equity; (c) intragenerational equity requires the correction of the asymmetrical distribution of the costs and benefits arising from the establishment of PAs; (d) the institutional features relevant to the environmental-economic issues being analysed should be considered; and (e) the decision-making process should be participatory and action oriented. The framework is applied to two case studies in Vanuatu. These applications detail (a) stakeholders??? views and trade-offs faced in relation to forest management (b) modes of participatory research and decision-making, (c) forms of compensation that may be adopted in correcting asymmetrical distributions of the costs and benefits of PAs, and (d) institutional influences on ecosystems use and implications for conservation projects. The institutional arrangements developed for the establishment of the PAs are presented. The application of this ecological economic framework has resulted in the formal establishment of one PA and the identification and assessment of five other PAs.

The objective of this thesis is to develop an ecological economic framework for the assessment and establishment of protected areas (PAs) that are aimed at conserving forests and biodiversity. The framework is intended to be both rigorous and relevant to the decision-making process. Constructivism is adopted as the paradigm guiding the research process of the thesis, after firstly examining also positivist philosophy and ???post-normal??? scientific methodology. The tenets of both ecological and environmental economics are then discussed. An expanded model of human behaviour, which includes facets derived from institutional economics and socioeconomics as well as aspects of neoclassical economics, is outlined. The framework is further developed by considering, from a contractarian view point, the implications of intergenerational equity for biodiversity conservation policies. The issues of intragenerational distribution and allocation are then considered. In this regard, cost-benefit analysis (CBA), as applied to the valuation of forests, PAs, and biodiversity, is critically reviewed. A participatory approach to decision-making, which may also include CBA, is then proposed. The resulting ecological economic framework may be thus summarised: (a) ecosystem use patterns should be chosen on the basis of their sustainability, distributional, and efficiency aspects; (b) systems of PAs should be established in order to achieve minimal intergenerational equity; (c) intragenerational equity requires the correction of the asymmetrical distribution of the costs and benefits arising from the establishment of PAs; (d) the institutional features relevant to the environmental-economic issues being analysed should be considered; and (e) the decision-making process should be participatory and action oriented. The framework is applied to two case studies in Vanuatu. These applications detail (a) stakeholders??? views and trade-offs faced in relation to forest management (b) modes of participatory research and decision-making, (c) forms of compensation that may be adopted in correcting asymmetrical distributions of the costs and benefits of PAs, and (d) institutional influences on ecosystems use and implications for conservation projects. The institutional arrangements developed for the establishment of the PAs are presented. The application of this ecological economic framework has resulted in the formal establishment of one PA and the identification and assessment of five other PAs.