Disa comprises 163 species, 131 of which occur in South Africa (SA). The genus is distributed across winter- and summer-rainfall areas, but few species transverse both climatic regions. Species are therefore regarded as winter-rainfall or summer-rainfall endemics - yet release their seeds in autumn, irrespective of provenance. Disa contributes 40 % of threatened Orchidaceae in SA, with half of the local species requiring conservation initiatives. In vitro seed germination is a potential conservation tool for producing large numbers of genetically diverse plants in relatively short periods. However, only 11 winter-rainfall Disa species are easily germinated ex situ. Studies were therefore undertaken on summer-rainfall taxa, which are ungerminated in vitro, in an effort to define their germination parameters. This thesis describes mechanisms that control germination in Disa and establishes practical propagation methods for seed culture. Two seed types occur in Disa; i) comparatively large, pale and pyriform seeds in members of the D. uniflora sub-c1ade, which populate streamside habitats under conditions of winter-rainfall maxima, and ii) smaller, variously brown and fusiform seeds in the remainder of the genus. Seed morphometrics distinguished seed types, although embryo dimensions were similar. Testa continuity, which is disrupted in the large seeds, also supported separation. Typically, small seeds are ungerminated in vitro, whilst large seeds germinated readily. Increased seed size did not necessarily impart increased germ inability, as several germinable, small-seeded species exist - being winter-rainfall species Attempts to establish in vitro germinability revealed that increased water availability and charcoal supplementation promoted germination in intractable species. The control of germination was therefore proposed as a trade-off between water availability and the presence of phyto-inhibitors - two features typical of seeds exhibiting water-impermeable dormancy. Three germinability categories were recognized; i) easily germinable species, ii) poorly germinable species through media manipulation, and iii) ungerminated species. Germination of immature seed in the absence of media modification was comparable to mature seed germination under modified conditions, providing evidence of the role of an impermeable seed testa in regulating germination. Testa impermeability in mature, small-seeded species was demonstrated using aqueous EVANS' blue dye and was linked to i) testa integrity and ii) increased levels of leachable phenolics (LPC) - which are hydrophobic and phytotoxic. In addition, this research revealed an impervious and elaborate embryo carapace in small seeds. Large-seeded species were highly permeable at dehiscence, with perforated testae and negligible LPC. Germinability was ultimately defined by a significant regression with LPC. Phenolic deposition increased exponentially with increasing seed maturity and reflected decreased permeability and the development of testa colouration. The testa precludes the use of viability stains such as nc and FDA, unless rendered permeable through scarification. This was achieved using NaOCI. Viability and germinability percentages did not correlate well for the small-seeded Disa species, indicating that i) the methods used to break dormancy are inadequate, ii) additional factors may be acting in concert with the testa to regulate germination and iii) that the determination of mature Disa seed viability is ineffective. As an alternative, the germination potential of immature seed was estimated as the ratio between the proportion of embryos stained with TTC and the proportion of seeds permeable to EVANS' blue. Attempts to relieve water-impermeable dormancy in Disa resulted in the formulation of a dual-phase protocol - with the specific aim of increasing water availability to the embryo. Dual-phase cultures comprised a solid, charcoal-rich medium overlaid with a reduced strength, liquid medium fraction of the same type. The solid fraction negated the influence of leached phenols and allowed protocorms to establish polarity, whilst the fluid fraction increased water availability. The dual-phase protocol allowed germination of nine summer-rainfall Disa species, usually in percentages that approximated their estimated germination potential. For the remaining species, germination is controlled by more complex factors. Large seeds are atypical in containing starch, the hydrolysis of which facilitated their rapid, autonomous germination. Small-seeded Disa species stored lipids and proteins and germinable species accumulated starch post-germination. The embryo protoplasts of all species contained appreciable amounts of soluble sugars, irrespective of germinability. However, decreased sucrose and increased fructose correlated significantly with decreased seed germinability. This study provides evidence of the nutritional value of mycotrophy, with endophytes liberating soluble carbohydrate and non-carbohydrate compounds upon lysis. However, few species were germinated symbiotically, suggesting that endophytes isolated from adult roots do not necessarily support germination in the same species. Similar endophytic fungi occur in Australian and Holarctic orchids. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2003.

The endemic landbirds of the Seychelles granitic islands have suffered considerable losses due to predation by introduced rats and cats and extensive habitat destruction. With less than 100 individuals, the Critically Endangered Magpie Robin Copsychus sechellarum Newton, faces the greatest risk of extinction. Translocations to three predator-free islands, Aride, Cousin and Cousine, have provided valuable opportunities for gaining insights into the ecology of the species. Of particular interest are links between the Magpie Robin, endemic skinks Mabvya spp., ground-living invertebrates and seabird colonies. Magpie Robin faecal pellet and skink gut content analysis demonstrated a high degree of dietary overlap between the species. A widespread exotic cockroach Pycnoscelus indicus was the favourite prey item for each species. However, behavioural observations and a dietary choice experiment indicated that there is no significant competition for food during the main seabird breeding season. Invertebrate sampling on Cousine identified 52 species which were available in all habitat types currently in use or those considered suitable for the Magpie Robin. Seabird and skink density counts on Cousine demonstrated the considerable magnitude of vertebrate organic food also available. Invertebrate sampling results on Cousin and Cousine were used to determine territory quality and the carrying capacity of each island for the Magpie Robin. While most seabirds are not breeding, skink survival depends on invertebrate abundance. M. wrightii weight declined throughout this period but that of M. sechellensis remained fairly stable. The data were insufficient to conclude that inter-specific competition for food exists between M. sechellensis and the Magpie Robin during this period. Further expansion of the Magpie Robin population depends on eradicating mammalian predators from other islands and maximising the potential carrying capacities of those already supporting the species. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1998.

Lake sturgeon, recently recommended to be listed as an endangered species under the Species at Risk Act, inhabit the various impoundments of the Winnipeg River system. Downstream passage through hydroelectric generating stations represents one of the major data gaps in our understanding of how hydroelectric development may be impacting lake sturgeon populations. Acoustic telemetry was used to investigate coarse-scale movements of juveniles, sub-adult and adults throughout the Slave Falls Reservoir, a 10 km long Winnipeg River impoundment, to assess patterns of downstream passage susceptibility and investigate fine-scale movements in the vicinity of the Slave Falls Generating Station. Movements of juveniles and sub-adults were generally restricted to areas of interconnected deep-water habitat, with movements through the shallow river narrows that sub-divide the Slave Falls Reservoir being rare. Adults did move through these narrows, albeit infrequently. Juveniles and sub-adults tagged in the lowermost section of the reservoir, as well as several adults tagged throughout the reservoir, were found to periodically utilize habitat immediately upstream of Slave Falls, where they would be susceptible to entrainment. Mean entrainment rates were estimated at 3.1% per year for adults tagged throughout the reservoir, and 17.9% per year for sub-adults tagged in the lowermost section of the reservoir. Fine-scale movement tracking revealed that three of eleven observed downstream passage events occurred via bottom-draw regulating gates, while another four events were also reasoned to have occurred via this route. The routes of the remaining four could not be determined. Eight of the eleven downstream passage events observed in this study were survived. While the survival of the remaining three fish could not be confirmed, it is highly likely that they also survived. Length-at-age analysis, supported by genetic methods, revealed that 23 of 151 (15.2%) of the lake sturgeon between 525 and 750 mm (fork length) captured in the 6 km stretch of river downstream of Slave Falls were fast-growing outliers, reasoned to have passed downstream through the Slave Falls Generating Station.

Wetland value is threatened by invasive plant species such as Reed Canarygrass (Phalaris arundinacea). Hence the research objectives of this project were to determine if reed canarygrass abundance has an effect on plant species diversity and assess the effectiveness of novel treatments on reed canarygrass control in a constructed wetland. Four treatments (mowing, herbicide, mowing plus herbicide, and a control) followed by broadcast seeding were applied to regulate growth of reed canarygrass. Principal components analysis, biodiversity measures, and ANOVA were used to identify community composition, quantify biodiversity values and identify treatment differences respectively. Results indicated differences in species composition between east and west blocks of the study site, reed canarygrass abundance appears to keep plant species diversity low, indigenous species were rare, and reed canarygrass was resistant to treatments.The results of this study are not surprising considering there is little evidence that treatments for reed canarygrass control are effective.

Infection with challenge virus standard-11 (CVS) strain, a laboratory fixed rabies virus strain, induces neuronal process degeneration in both in vivo and in vitro models. CVS-induced axonal swellings of primary rodent dorsal root ganglion neurons are associated with 4-hydroxy-2-nonenal staining indicating a critical role of oxidative stress. Mitochondrial dysfunction is one of the most important causes of oxidative stress. We hypothesized that CVS infection induces mitochondrial dysfunction leading to oxidative stress. We investigated the effects of CVS infection on several mitochondrial parameters in different cell types. CVS infection increased electron transport chain capacity, Complex I and IV activities, but did not affect Complex II-III, citrate synthase, and malate dehydrogenase activities. CVS maintained normal oxidative phosphorylation capacity and proton leak, indicating a tight mitochondrial coupling. Possibly as a result of enhanced Complex activity and efficient coupling, a high mitochondrial membrane potential was generated. CVS infection reduced the intracellular ATP level and altered the cellular redox state as indicated by high NADH/NAD+ ratio. CVS infection was associated with a higher rate of hydrogen peroxide production. We conclude that CVS infection induces mitochondrial dysfunction leading to ROS overgeneration, oxidative stress and neuronal process degeneration.

I investigated how orchid biology, floral morphology, and diversity of surrounding floral and pollinator communities affected reproductive success and hybridization of Cypripedium candidum and C. parviflorum. Floral dimensions, including pollinator exit routes were smallest in C. candidum, largest in C. parviflorum, with hybrids intermediate and overlapping with both. This pattern was mirrored in the number of insect visitors, fruit set, and seed set. Exit route size seemed to restrict potential pollinators to a subset of visiting insects, which is consistent with reports from other rewardless orchids. Overlap among orchid taxa in morphology, pollinators, flowering phenology, and spatial distribution, may affect the frequency and direction of pollen transfer and hybridization. The composition and abundance of co-flowering rewarding plants seems to be important for maintaining pollinators in orchid populations. Comparisons with orchid fruit set indicated that individual co-flowering species may be facilitators or competitors for pollinator attention, affecting orchid reproductive success.

In the face of ever increasing atmospheric CO₂ a better understanding of soil properties and processes and the effect of management practices, such as the application of nitrogen fertilizer is of importance and could potentially improve our ability to sustainably manage forestry systems. With that in mind this study was conducted in order to investigate the effects of tree species and fertilization on soil carbon and the soil microbial community. To this end, soil from fertilized and unfertilized plots at Berwick forest, under stands of Pinus radiata and Sequoia sempervirens at Hanmer and under six different tree species at Holt forest was sampled. Two glasshouse pot trials were established using soil collected from the Hanmer and Berwick forest sites and seedlings of Pinus radiata, Sequoia sempervirens, and Eucalyptus nitens were grown. Soil properties were determined from both the field sites and pot trials including soil organic matter, carbon, nitrogen, and microbial biomass by chloroform fumigation extraction. Biolog ecoplates were used to determine the relative differences in diversity based on substrate utilization patterns of the soil microbial communities in soil sampled from the glasshouse pot trials. Soil microbial biomass carbon, nitrogen and the ratio of microbial biomass carbon:nitrogen differed significantly between Pinus radiata and the other tree species sampled at Holt forest. Significant effects of fertilization and tree species on soil carbon and microbial biomass were observed in both pot trials. Soil carbon differed significantly between Eucalyptus nitens and both Pinus radiata and Sequoia sempervirens in the first pot trial and relative to both, E. nitens contributed significantly more carbon. No significant effect of either fertilization or tree species on the catabolic diversity of the soil microbial community in both glasshouse pot trials was observed. The results demonstrated the effects that fertilization and tree species can have. Particularly notable was the short-time period in which tree species effects became apparent coupled with the absence of any aboveground inputs to the soil.

Biological invasions pose a serious threat to biodiversity world-wide. Through various means, such as competition or predation, invaders can radically change species composition and the functioning of native ecosystems. Even though our understanding of the mechanisms underlying invasion success is improving, there is still a lack of knowledge on the response of native species under pressure from invasion. This study adds to existing knowledge on the responses of a native species to invasion by non-indigenous species. Pyura pachydermatina is a native ascidian in the southeast coast of New Zealand currently under pressure from increased urbanisation and invasion by other ascidian species. The reproductive strategies employed by P. pachydermatina are investigated and the role of these strategies to increase its resistance to invasion are assessed. A population study on the status of P. pachydermatina around the Banks Peninsula was carried out in Camp Bay, Pigeon Bay, and Wainui. Spawning experiments using P. pachydermatina and gonad histology were done regularly during the one year study period to assess its ability to self-fertilise and determine its reproductive period. In addition, predation experiments were carried out to assess the susceptibility of P. pachydermatina early life stages to two amphipod predators. The surveys indicated that the populations of P. pachydermatina in the three sites are different from one another. Wainui has on average the largest individuals of P. pachydermatina and Camp Bay, the smallest. Abundance of P. pachydermatina was highest in Pigeon Bay and lowest in Wainui. The three life stages of Pyura pachydermatina; recruits, juveniles, and adults, were present in all sites at all seasons. The spawning experiments confirmed the species’ ability to self-fertilise and that it has a year-round spawning period. The two amphipod predators, Jassa marmorata and Caprella mutica, were efficient in consuming the egg and larval stages of P. pachydermatina, but did not feed on the settlers. Year-round reproduction and the ability to self-fertilise potentially give P. pachydermatina increased resistance to the effects of urbanisation and invasion. This population study suggested that the species is thriving around the Banks Peninsula. This, combined with previous studies on the non-indigenous ascidian Styela clava that stated the static or declining populations of the potential invaders, gives a positive outlook for the native species for the future. I suggest the use of genetic techniques to assess, in more detail, the population structure and dispersal potential of this native species. I also suggest constant monitoring of native species is required to keep up to date with the current status of the species, which will in turn help management decisions should regional spread of the Lyttelton S. clava invasion occur in the future.

Brachyglottis (Asteraceae) is a genus of approximately 30 species in the Brachyglottidinae, a recently recognised sub-tribe of tribe Senecioneae. Within Brachyglottis is a clade of five species of rosette-forming herbs: B. bellidioides, B. haastii, B. lagopus, B. southlandica and B. traversii. A sixth species, B. saxifragoides, has recently been synonymised with B. lagopus. The rosette-Brachyglottis have historically been recognised as a taxonomically problematic group because species overlap in both morphology and geographical distribution. A recent molecular study of rosette-Brachyglottis using AFLP data indicated that genetic distances among populations of rosette-Brachyglottis in the South Island appear to be correlated with geographical distance between populations rather than taxonomic identification. This is problematic as the currently described rosette-Brachyglottis species have overlapping ranges which implicitly hypothesises reproductive barriers other than geographic distance. We conducted an investigation into the species delimitation of rosette-Brachyglottis with the aim of answering two related questions: Does the current delimitation of rosette-Brachyglottis accurately reflect patterns of genetic similarity? Do the patterns of genetic structure in rosette-Brachyglottis support the presence of multiple biological species? A total of 46 populations of rosette-Brachyglottis were represented in this study. Herbarium specimens collected from these populations were identified following the taxonomic treatment of Allan (1961). Twenty one discrete and numerical morphological characters were measured from herbarium specimens including those collected for this study and previously collected herbarium specimens. Morphological dissimilarity of 354 herbarium specimens was investigated by performing a PCoA on Gower’s pairwise morphological distances among individuals. The pattern of genetic similarity was explored using DNA fragment length variation in nine markers for 273 individuals and this resulted in a total 177 unique alleles. Bayesian clustering analysis was performed on this data set using STRUCTURE, in addition, pairwise genetic distances were calculated among individuals and populations using Jaccard and Nei’s dissimilarity coefficient’s respectively. Jaccard genetic distances among individuals were analysed using PCoA and Nei’s genetic distances among populations were analysed using a Neighbour-Net analysis. The relationship between pairwise genetic and geographic distances among populations was analysed using a combination of linear regression and a Mantel Test. The pattern of morphological similarities among specimens was generally congruent with the currently delimited species in rosette-Brachyglottis. However, many morphologically intermediate specimens confound the recognition of distinct morphological entities. Comparison of patterns of genetic similarity and the current morphologically-based species delimitation showed that the delimitation does not accurately reflect the genetic structure of rosette-Brachyglottis. Furthermore, patterns of genetic dissimilarity did not indicate discrete genetic groups at the individual or population levels. The finding of incongruence between patterns of genetic and morphological similarity and absence of morphologically or genetically discrete groups suggests that rosette-Brachyglottis are best considered a single, yet morphologically diverse, biological species. In addition genetic structure within this species appears to be primarily driven by geographical isolation.

Biometric identification is a pattern recognition based classification system that recognizes an individual by determining its authenticity using a specific physiological or behavioural characteristic (biometric). In contrast to number of commercially available biometric systems for human recognition in the market today, there is no such a biometric system for plant recognition, even though they have many characteristics that are uniquely identifiable at a species level. The goal of the study was to develop a plant species biometric using both global and local features of leaf images. In recent years, various approaches have been proposed for characterizing leaf images. Most of them were based on a global representation of leaf peripheral with Fourier descriptors, polygonal approximations and centroid-contour distance curve. Global representation of leaf shapes does not provide enough information to characterise species uniquely since different species of plants have similar leaf shapes. Others were based on leaf vein extraction using intensity histograms and trained artificial neural network classifiers. Leaf venation extraction is not always possible since it is not always visible in photographic images. This study proposed a novel approach of leaf identification based on feature hierarchies. First, leaves were sorted by their overall shape using shape signatures. Then this sorted list was pruned based on global and local shape descriptors. The consequent biometric was tested using a corpus of 200 leaves from 40 common New Zealand broadleaf plant species which encompass all categories of local information of leaf peripherals. Two novel shape signatures (full-width to length ratio distribution and half-width to length ratio distribution) were proposed and biometric vectors were constructed using both novel shape signatures, complex-coordinates and centroid-distance for comparison. Retrievals were compared and the biometric vector based on full-width to length ratio distribution was found to be the best classifier. Three types of local information of the leaf peripheral (leaf margin coarseness, stem length to blade length ratio and leaf tip curvature) and the global shape descriptor, leaf compactness, were used to prune the list further. The proposed biometric was able to successfully identify the correct species for 37 test images (out of 40). The proposed biometric identified all the test images (100%) correctly if two species were returned compared to the low recall rates of Wang et al. (2003) (30%, if 10 images were returned) and Ye et al. (2004) (71.4%, if top 5 images were returned). The biometric can be strengthened by adding reference images of new species to the database, or by adding more reference images of existing species when the reference images are not enough to cover the leaf shapes.