The seed of Santalum spicatum is rich in a fixed oil (59% by weight), which is characterised by a high percentage of acetylenic, ethylenic ximenynic acid (35% of total fatty acids). A number of important aspects of the seed fixed oil, its composition in developing seeds, its triacylglycerols molecular species in the oil, the nutrition and toxicity of the oil feeding, and the possible bioactivity of ximenynic acid in mice were investigated.The identification of cis and trans isomers of ximenynic acid in the seed oil, and the metabolite of ximenynic acid in mouse liver lipid fractions were achieved using 2-amino-2-methyl-1-propanol to form 2-substituted 4,4-dimethyloxazoline derivatives, which were analysed by gas chromatography with mass spectrometric detection.Changes in proximate and fatty acid composition were investigated in developing seed collected weekly from about seven days after flowering to maturity. It was determined that moisture and carbohydrate contents decreased significantly during the development sequence, while fixed oil content increased from 0.3% to 50% (by weight) with seed development. A corresponding increase in the proportions of both oleic and ximenynic acids occurred suggesting a precursor/product relationship. Mature seed collected from different locations in Western Australia showed minor differences in characteristics and lipid composition, which may have been influenced by geographical origin and harvesting year of samples.The lipid components from the seed oil were separated using thin-layer chromatography and the individual triglyceride bands were characterised by high performance liquid chromatography and gas chromatography using flame ionisation and mass spectrometric detection after removal from the plate. The triximenynin (trisantalbin) band showed no other contaminating fatty acids and was obtained in a relatively pure state.A ++ / nutrition and toxicity study was performed by feeding a semi-synthetic diet containing sandalwood seed oil to a level of 15% of total energy content to a group of mice for one month and another group for two months. The most significant effect of sandalwood seed oil ingestion when compared with a standard lab diet (5% fat, by weight) and a canola oil-enriched diet (15% fat, by weight) was an apparent reduction in body weight gain, which may be the effect of ximenynic acid as a growth retardant. Serum aspartate aminotransferase levels were determined in the mice as an indicator of hepatotoxicity. These levels were higher in mice fed the sandalwood seed oil diet than those fed the standard lab diet, suggesting that ximenynic acid may affect liver-specific enzyme activity. Analysis of the total lipid fatty acids of various tissues and organs of mice showed only a low incorporation of ximenynic acid into the general tissues (0.3-3% by weight), and its absence in the brain.This study suggests a few health benefits from consumption of large quantities of sandalwood seed oil in the diet. These include a low lipid content in blood, heart, muscle, increase in the 16:1/16:0 and 18:1/18:0 ratios, production of increased levels of 18:1 (n-9) and docosahexaenoic acid, and decreased levels of arachidonic acid in certain tissues. There were no specific pathological, morphological or mortality changes observed in the mice.Sandalwood seed may be both a food and a medicine.

[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.