This thesis presents an investigation into the chemistry of 6 substituted 5, 6-dihydro-a-pyrone compounds. A comprehensive review of these compounds was published in 1989 and the subsequent literature is covered in an updated review presented below. Eight 6-substituted 5,6-dihydro-a-pyrone metabolites from three different South African plant species Cryptocarya latijolia, Syncolostemon densiflorus, and Syncolostemon argenteus have been the subject of structural and stereochemical investigations. The absolute stereochemistry of the known compound "triacetate" from C. latijolia has been established as 6R-[2R,4S,6S-(triacetyloxy)heptylJ-5,6-dihydro-2H-pyran-2-one (74) using CD and acetonide formation with subsequent application of the modified Moshers method. The absolute stereochemistry of the related metabolite "diacetate", also from C. latijolia, has been assigned as 6R-[2S,4S-diacetyloxypentylJ-5,6-dihydro-2H-pyran-2-one (76). In addition, the outstanding stereochemistry at C-5' in syndenolide, from S. densiflorus, followed from conversion to its diacetonide and subsequent NMR analysis. Syndenolide is therefore 6R-[5S-(acetoxy)-IR,2R,3S-(trihydroxy)-heptylJ-5,6- dihydro-2H -pyran-2-one. The genus Syncolostemon has proved to be a rich source of a-pyrone compounds and the chemistry of S. argenteus, not investigated previously, was examined as part of an ongoing search for new 5,6-dihydro-a-pyrones. The study yielded five new a-pyrone natural products, synargentolide A-E. The structure of synargentolide A (82) has been assigned as 6R[4R,5R,6S-triacetyloxy-lE-heptenylJ-5,6-dihydro-2H-pyran-2-one using CD and NMR techniques. The structures of synargentolide B (87), C (92) and E (94) also followed from a detailed NMR analysis and the stereochemistry tentatively assigned based on CD and NMR data. Synargentolide D (93) was thermally unstable, and a paucity of material prevented stereochemical investigations, however the structure was determined from initial NMR analysis. The marine molluscs of the genus Siphonaria have only become the subject of chemical studies in the last fifteen years. These molluscs characteristically produce polypropionate type natural products. A review of Siphonarian polypropionate metabolites containing a pyrone functionality is presented. Examination of an endemic South African species Siphonaria serrata yielded one novel polypropionate metabolite containing a ),-pyrone functionality, siserrone A (131). The structure of this compound was unambiguously established using standard NMR experiments. The relative stereochemisty of the hemi-ketal moiety was assigned from a careful analysis of the ROESY NMR spectrum and the stereochemisty of the acyclic portion determined from a comparison of the 13C and 'H NMR data of a degradation product with the corresponding data of a synthetic compound. It was also established that the modified Moshers method could not be used to determine the absolute stereochemistry of the secondary hydroxy I substituent at C-11. The absolute stereochemistry of 131 was thus assigned in accordance with the proven stereochemistry of Siphonarian metabolites.

The reproductive strategies of marine invertebrates have been related to various aspects of both their ecology, and their phylogenetic history. It has been the purpose of this thesis to try and separate these components among Siphonaria, a group of marine pulmonates. The taxonomy of these species is revised and I conclude that nine species are valid. All species deposit benthic egg masses and development may be either direct (S. anneae, S. compressa, S. dayi, S. nigerrima, S. serrata and S. tenuicostulata) or planktonic (S. capensis, S.concinna and S. oculus). Data on distribution and life-history relating to mode of larval development is then presented for 26 species of Siphonaria worldwide. Fifteen species are direct developers, nine are planktonic developers and a further two appear to have a dual developmental capacity, retaining both the velar swimming apparatus of a planktonic developer and the crawling foot of a direct developer. Direct developing species hatch from larger egg capsules, and generally occur higher on the shore than planktonic developers. Worldwide, planktonic developers are more widespread than direct developers, and individual planktonic species may have a greater latitudinal range. In most S~honaria subgenera, mode of larval development appears to be constant, although two subgenera (Patel/opsis and Sacculosiphonaria) include both developmental types. Locally, the intertidal zonation of three sympatric species (S. capensis, S. concinna and S. serrata) does not support a model which predicts direct development on the high shore and planktonic development on the low shore. However, distributions do correspond to particular intertidal microhabitats, and while there may be no, direct relationship between mode of larval development and intertidal height, the physical structure of egg masses, and the microhabitats used for spawning appear adaptive with regards to desiccation in the intertidal. S. concinna (planktonic development) and S. serrata (direct development) occur in similar microhabitats and are likely to be under similar selection pressures. In having different modes of larval development, there appears to be more than one optimal solution in a particular selective regime. In addition, both species seem to apportion similar amounts of energy to reproduction for each spawning episode, and also annually suggesting an optimum allocation of resources to reproduction. Genetic investigations using PolyAcrylamide Gel Electrophoresis (PAGE) confirm the status of the southern African species initially described, and indicate greater genetic variability associated with planktonic developing species than direct developing species. The systematic relationships revealed by DNA fingerprinting support the current classification systems, and also have implications with regards the evolution of larval development: direct development may be the plesiomorphic condition in, and among, some Siphonaria groups. There are both phylogenetic and adaptive explanations for the distribution of reproductive mode among benthic marine invertebrates. An evolutionary question, however, is not just a matter of either adaptation or constraint, it is a combination of these. Both contribute to the distribution of developmental mode among Siphonaria.

Physiological response to environmental change is examined in species of the intertidal limpets, Patella (Prosobranchia) and Siphonaria (Pulmonata). Characteristics of heart beat were determined using impedance pneumography and these are described. Heart rate of P. granularis is related to temperature and body size, and cardiac arrest in this limpet is apparently stress-related. Siphonaria oculus may exhibit a temperature-independent, extreme, and often prolonged bradycardia (<10 beats/min). When measured shortly after aerial exposure, heart rate and oxygen consumption of the above limpet species are closely correlated. The relationships of aerial oxygen consumption with body weight and ambient temperature were determined for the above high shore species of limpet. Both have low aerial rates of oxygen consumption relative to low shore limpet species, and their QlO values decrease with increasing temperature. Diel field recordings of heart rate of S. oculus, taken during summer and winter, suggest absence of temperature acclimation, and this was also shown for oxygen consumption at high aerial temperatures (30°C) in laboratory experiments. Oxygen consumption of P. granularis is partially temperature compensated temperature acclimation). In air, even though S. oculus loses water faster, it shows greater tolerance of water loss and survives longer than P. granularis. Prolonged aerial exposure of S. oculus leads to depression of heart rate and of V02 (down to 18% of the pre-exposure rate), responses interpreted as representing adaptive metabolic rate depression. In P. granularis aerial heart rate remains constant and V02 never falls below IX 38% of pre-exposure rate. This reduction in V02 in air is considered as being stress-related, resulting from impairment of oxygen uptake. In declining oxygen tension S. capensis shows a better capacity for oxyregulation than P. granularis. On exposure to hypoxia, sand-inundation and hyposalinity, S. capensis may show typical bradycardia ( <10 beats/min), suggesting depression of aerobic metabolism, and on return to pre-exposure conditions there is no overshoot of heart rate, suggesting absence of oxygen debt. The effect of hypoxia, sand-inundation and hyposalinity on heart rate of P. granularis is variable; this becomes depressed and is often interspersed with extended cardiac arrest. When conditions are normalized, this species exhibits a clear overshoot of heart rate. The significance of differences in physiological response between patellid and siphonariid limpets is discussed with regard to their habitat segregation, particularly in the upper-shore zone (open rock and tidal pools) and sand-inundated rock substrata, where only siphonariid limpets may be found. While not previously characterized in marine gastropods, metabolic rate depression by Siphonaria, through facilitating isolation and conserving food reserves, is suggested as a key factor determining their distribution in physico-chemically more extreme and variable intertidal habitats.

Dispersal rates are known to play a fundamental role in establishing the genetic structure found within a species. Dispersal in marine benthic invertebrates is largely dependent on life history strategy. The presence of pelagic larvae will potentially result in high dispersal within a species, while direct development limits dispersal. This study used two intertidal pulmonate limpet species to investigate the relationship between dispersal potential and genetic structure of populations. Siphonaria capensis produces benthic egg masses, which release pelagic larvae. These float in ocean currents before settling and metamorphosing into adults. S. serrata reproduces by direct development. The larvae hatch as completely metamorphosed juveniles after developing in an egg case not far from the parent. The high dispersal capacity of S. capensis pelagic larvae should result in high levels of within-population variation since individuals within a particular population may originate from several different populations. This should lead to high levels of gene flow along the coast and to low genetic differentiation among populations. On the other hand, the low dispersal potential of S. serrata should cause low levels of genetic variation within populations, as relatives will remain close to one another. Low levels of gene flow and high genetic differentiation among populations are expected in this species. In dealing with the relationship between life history strategy and population genetic structure in these two species, a series of indirect techniques was utilised. Multivariate analyses of the morphological variation within each species showed that variation in size was similar between the two species and largely governed by environmental factors. Variation in shell shape was largely under genetic control and supported the predictions made for each species. Variation in total proteins was relatively uninformative in terms of examining the differences between the two species. Allozyme analysis and mtDNA sequencing clearly showed differences between the species in terms of their population genetic structure. High levels of gene flow were found within S. capensis. This was strongly influenced by ocean currents, with the close inshore Agulhas current along the Transkei coast contributing to high levels of dispersal and hence gene flow. Low levels of gene flow occurred within S. serrata, resulting in low within-population variation and high among-population differentiation.

published_or_final_version / Ecology and Biodiversity / Doctoral / Doctor of Philosophy

In recent decades the emergence of marine polypropionate natural products as compounds of diverse structural complexity and intriguing biological activity has influenced the advancement of asymmetric synthesis and predicated detailed studies of marine ecology. The introductory chapter of this thesis explores the nature of marine natural products, including their structure, biological activity and biosynthesis. Additionally, a brief review of the aldol reaction is presented. This well established biomimetic chemical transformation underpins polyketide synthesis and was utilised extensively in the research contributing to this dissertation. Chapter Two describes the first asymmetric total synthesis of the two marine polypropionates isolated from specimens of Siphonaria australis by Hochlowski et al. in 1984. Spectroscopic analysis revealed hemiacetal 22 and ester 23 to be identical to the secondary metabolites extracted from the marine pulmonate. The synthetic approach to hemiacetal 22 utilised lactate derived ketone (S)-67 to control the configuration of the C7 and C8 stereocentres and involved the discovery of a mild protocol for the synthesis of trimethylsilyl enol ether 109, which was employed for a Mukaiyama aldol homologation reaction. Additionally, ester 23 was synthesised from hemiacetal 22 via a retro-Claisen fragmentation. The retro-Claisen approach utilised in the synthesis of ester 23 was extended in Chapter Three to serve as the pivotal transformation in an attempted total synthesis of the unusual marine polypropionate dolabriferol (30). The strategy toward dolabriferol (30) involved an iterative homologation of chiral ketone (S)-67 to install all but one of the requisite stereocentres in the natural product. Chemoselective deprotection of acyclic precursor 160 gave the elaborate 2,4,6-trioxaadamantane 167, whose participation as a protecting group mimic lead to the formation of ester 169 after reaction of the polycycle 167 with base. The synthesis of ester 169, which represents a direct precursor to dolabriferol (30), was achieved in 16 steps with an overall yield of 24%. Unfortunately, a robust protecting group on ester 169 prohibited a synthesis of dolabriferol (30), but intriguingly in one deprotection of ester 169 with aqueous hydrofluoric acid, spiroacetal 172 was isolated. Chapter Four describes the first total synthesis of cytotoxic marine polypropionate auripyrone A (78) and establishes the absolute configuration of this important natural product as that depicted for compound 78. The requisite C8-C12 stereopentad of auripyrone A (78) was formulated from Evans� dipropionate equivalent 53 in a double stereodifferentiating aldol reaction, followed by syn-reduction to give diol 206. Differentiation of the secondary alcohols in compound 206 was achieved by migration of the PMB protecting group and protection at C11 with the requisite acyloxy group of auripyrone A (78). Differential protection was critical to achieving selective spiroacetalisation to afford the unique spiroacetal dihydropyrone core of the natural product. The utility of LiHMDS for highly selective double stereodifferentiating aldol homologations of sensitive fragments is also discussed. This mild aldol protocol was pivotal to forming the carbogenic skeleton of auripyrone A, in particular, elaborate adduct 278.

Habitat fragmentation is a fundamental process that determines trends and patterns of distribution and density of organisms. These patterns and trends have been the focus of numerous terrestrial and marine studies and have led to the development of several explanatory hypotheses. Systems and organisms are dynamic and no single hypothesis has adequately accounted for these observed trends. It is therefore important to understand the interaction of these processes and patterns to explain the mechanisms controlling population dynamics. The main aim of this thesis was to test the effect of patch size and isolation on organisms with different modes of dispersal. Mode of dispersal has previously been examined as a factor influencing the effects that habitat fragmentation has on organisms. Very few studies have, however, examined the mode of dispersal of marine organisms because it has long been assumed that marine animals are not directly influenced by habitat fragmentation because of large-scale dispersal. I used two co-occurring species of siphonariid limpets with different modes of dispersal to highlight that not only are marine organisms affected by habitat fragmentation but that they are affected in different ways. The two species of limpet, Siphonaria serrata and Siphonaria concinna, are found within the same habitat and have the same geographic range along the South African coastline, however, they have different modes of dispersal and development. The effect of patch size on organism density has been examined to a great extent with varied results. This study investigated whether habitat patch size played a key role in determining population density and limpet body sizes. The two species are found on the eastern and southern coasts of South Africa were examined across this entire biogeographic range. Patch size was found to have a significant effect on population density of the pelagic developer, S. concinna, but not the direct developing S. serrata. Patch size did play a role in determining limpet body size for both species. S. concinna body size was proposed to be effected directly by patch size whilst S. serrata body size was proposed to be affected indirectly by the effects of the S. concinna densities. The same patterns and trends were observed at five of the seven examined regions across the biogeographic range. The trends observed for S. concinna with respect to patch size conform to the source-sink hypothesis with large habitat patches acting as the source populations whilst the small habitat patches acted as the sink populations. Many previous studies have focused on the effects of habitat patch size at one point in time or over one season. I tested the influence of habitat patch size on the two species of limpets over a period of twelve months to determine whether the trends observed were consistent over time or whether populations varied with time. S. concinna showed a consistently significant difference between small and large patches; whilst S. serrata did not follow a consistent pattern. The mode of dispersal for the two limpets was used to explain the different trends shown by the two species. This examination allowed for the determining of source and sink populations for S. concinna through the examination of fluctuations in limpet body sizes and population densities at small and large habitat patches over twelve months. The direct developing S. serrata trends could not be explained using source-sink theory, as populations were independent from one another. S. serrata demonstrated body size differences at small and large patches which, may be explained by interspecific and intraspecific competition. Habitat isolation is known to play an important role in determining the structure of assemblages and the densities of populations. In this study the population density of the pelagic developing S. concinna showed a weak influence of degree of isolation whilst that of the direct developing S. serrata did not, which may be because of habitat patches along the South African coastline not having great enough degrees of isolation. The population size-structure was influenced directly influenced by isolation for S. concinna, whilst the different population size structure for S. serrata may be explained by assemblage co-dependence. The mode of dispersal showed effects on the relationship of population density and population size-structure with habitat size and isolation. This study indicates the importance of investigating patterns and processes across a range of spatial and temporal scales to gain a comprehensive understanding of factors effecting intertidal organisms.