The intertidal red algal genus Porphyra is found on rocky shores worldwide. In the Northern Hemisphere the genus is well studied but there is a paucity of data on southern hemisphere Porphyra and even less on New Zealand Porphyra. The species� taxonomy has been undergoing revision since the late 1990�s, when it was discovered that the main species P. columbina and P. lilliputana reported for New Zealand were a combination of several endemic species. These species are found from the low to the high intertidal watermark; hence they are exposed to fluctuating stresses such as desiccation, temperature, high light and UV radiation. Algae have evolved a number of mechanisms to adapt to naturally changing increasing abiotic conditions, such as accumulation of screening pigments and changes in antioxidant metabolism during light stress. For terrestrial plants, polyamines (small aliphatic amines) have been shown to be involved in protecting cells from damage under conditions of stress including UV-B radiation; such mechanisms have yet to be identified in algae. The overall aim of this study was to determine the importance of cellular processes in shaping the community structure of Porphyra on a wave-exposed shore on the east coast of the South Island, New Zealand. Porphyra distribution and community structure was assessed by regular monthly monitoring of presence and absence of Porphyra along four transect lines at the site. Enviromnental information was recorded to determine the effects of temperature, light, UV radiation, humidity and wind on Porphyra�s spatial and temporal distribution. Regular tissue samples were taken for species identification by the application of primers, which were specifically designed during this study. P. cinnamomea and Porphyra spec. "ROS 54" were identified as dominant species present almost throughout the year with a pronounced maximum in presence during late winter and spring, and some weeks of absence during April or May. The two dominant species were recorded from the low to the high intertidal shore, but the mid intertidal was identified as the preferred habitat. Other species that were found were rare and only present for a few months in a very restricted area. It was hypothesised that free radical generation and antioxidant metabolism are associated with desiccation tolerance in Porphyra. An attempt was made to investigate the impact of desiccation stress on Porphyra. The extraction process of antioxidants was problematic and no reproducible results could be obtained. It was attempted to investigate the spatial distribution of spores and conchocelis of different Porphyra species in the field, and determine if those found at Brighton Beach are species-specific in their morphology. This indicated that the two main Porphyra species at Brighton Beach not only prefer to occupy the same habitat but that they also have a morphologically similar conchocelis phase. Mechanisms on a cellular level such as polyamine metabolism affected by environmental (abiotic) stresses are related to the alga�s ability to adapt to stress and therefore can have an effect on Porphyra�s distribution along the shore and its presence throughout the year. The depletion of the ozone layer has become an important issue as the effects of increased UV radiation on the environment, especially the intertidal habitat, are revealed. Marine macrophytes possess the main three. polyamines: putrescine, spermidine and spermine of varying levels. For the few species studied, Rhodophyta generally contain higher levels of polyamines than Chlorophyta, while polyamine levels for the one heterokontophyte analysed were between Chlorophyta and Rhodophyta. Levels of the three most common polyamines (putrescine, spermidine, spermine) were determined in P. cinnamomea under controlled UV exposure. Tissue discs were exposed to visible light (PAR), PAR and UV-A or PAR, UV-A and UV-B radiation. Discs exposed to PAR and PAR and UV-A showed little change in polyamine levels over a six day trial period, while discs exposed to PAR, UV-A and UV-B showed a significant increase in free, bound soluble and bound insoluble polyamines over the same period of time. Correspondingly levels of ADC and ODC, two enzymes involved in polyamine synthesis, were measured. ODC levels changed little while ADC levels increased significantly during UV-B treatment, indicating that under UV-B stress polyamines are mainly synthesized via the ADC pathway. The experimental set-up and process of this study has not been applied in macroalgal polyamine research and results obtained are the first indication that increased levels of polyamines are involved in protection and/or protection mechanisms in macrophytic algae to prevent UV-B damage.

The oceans represent a wealth of biological diversity where many species remain to be discovered and described. Among seaweeds, a paucity of morphological features by which to differentiate species means that many genera harbour overlooked or cryptic species. Fucus, Porphyra and Ulva are three common genera of marine intertidal algae and all include species that are particularly difficult to distinguish morphologically. DNA barcoding has been championed as a revolutionary tool for species identification and discovery and applying this tool to algae was a logical step due to the difficulty of morphological identification of many algal species. This thesis is part of a significant initiative aimed at identification and discovery of all species of seaweeds in Canadian waters, using a DNA barcoding approach. The original concept of DNA barcoding relied on comparing the 5’ region of the mitochondrial cytochrome c oxidase 1 (COI-5P) gene among animal species. In this study, DNA barcoding with COI-5P was applied to the brown algal genus Fucus and worked as well as any other marker to assign morphologies to known species. The DNA barcoding results also uncovered substantial phenotypic diversity in Pacific F. distichus. Results were confirmed by comparison with sequences of the nuclear internal transcribed spacer region (ITS). For Porphyra, COI-5P DNA barcoding was compared with species identification using the chloroplast large rubisco subunit (rbcL) and the Universal Plastid Amplicon (UPA) in a floristic survey of Canadian Porphyra species. Two new species were discovered and described (Porphyra corallicola and Porphyra peggicovensis), and P. cuneiformis was synonymized with P. amplissima. The COI-5P emerged as the best marker for species discrimination despite difficulties with primer universality. To aid in choosing a marker for DNA barcoding for green algae, the universality and species discriminatory power of the rubisco large subunit (rbcL) (considering the 5’ and 3’ fragments independently), the UPA, the D2/D3 region of the nuclear large ribosomal subunit (LSU-D2/D3) and the ITS were evaluated. While the rbcL-3P highlighted several cryptic species, and worked well to distinguish Ulva species, more research is needed to recommend a marker for DNA barcoding generally in marine green macroalgae.

Porphyra abbottiae Krishnamurthy (Rhodophyta) is an intertidal red alga harvested by a number of coastal First Nations in British Columbia. The Kwakwaka’wakw have a long history of harvesting P. abbottiae as food and medicine, reflected in the language, songs and stories of the Kwakwaka’wakw oral tradition. Harvesting and drying practices for this alga have undergone changes with the introduction of new technologies and a decrease in time available for seaweed harvesting. The adoption of timesaving equipment into the seaweed harvest has given harvesters the flexibility to work around constraints including work and school obligations, tides, long distances to harvesting sites, and unpredictable weather conditions. Harvesting and drying practices reflect a thorough understanding of the lifecycle, biology, and ecology of P. abbottiae. Timing of the harvest during the seasonal round optimizes the taste and texture of P. abbottiae fronds while avoiding the seaweed in its reproductive stage. Songs and taboos associated with the harvest promote safety and efficiency while harvesting the seaweed. Concerns about potential contamination of edible seaweed led to the second part of this research: testing for metal contamination. Inductively coupled plasma mass spectrometry analysis for selected metals and trace elements revealed the presence of arsenic, cadmium, lead, and mercury in Porphyra abbottiae sampled from the southern Queen Charlotte and Broughton Straits. Mercury concentrations fell below the detection limit of 0.01 ng/mL in 28 of 112 samples. Calcium was the most abundant element measured, averaging 1445 mg/kg dry seaweed. The remaining metals, in decreasing order of concentration, are: Fe>As>Zn>Mn>Cu>Cd>Pb>Cr>Co>Se>Hg. Copper-zinc (r=0.835) and copper-lead (r=0.948) concentrations are significantly correlated (p<0.05), suggesting selective uptake of these elements. PCA analysis suggests that the location of harvesting sites within specific water channels is influencing metal concentrations. Hazard quotients calculated using guidelines set by Health Canada and the World Health Organization revealed that, among the suite of elements surveyed, arsenic followed by cadmium ranked the highest in relative risk for consumers of P. abbottiae. An average 60 kg adult consumer can safely consume approximately 9.4 g dried seaweed per day and not exceed tolerable upper intake limit guidelines. In conclusion, Porphyra abbottiae can be eaten in moderation with minimal risk of chronic metal contamination. Kwakwaka’wakw consumers can also benefit from cultural reconnection with this important traditional food. / Graduate

The intertidal red algal genus Porphyra is found on rocky shores worldwide. In the Northern Hemisphere the genus is well studied but there is a paucity of data on southern hemisphere Porphyra and even less on New Zealand Porphyra. The species� taxonomy has been undergoing revision since the late 1990�s, when it was discovered that the main species P. columbina and P. lilliputana reported for New Zealand were a combination of several endemic species. These species are found from the low to the high intertidal watermark; hence they are exposed to fluctuating stresses such as desiccation, temperature, high light and UV radiation. Algae have evolved a number of mechanisms to adapt to naturally changing increasing abiotic conditions, such as accumulation of screening pigments and changes in antioxidant metabolism during light stress. For terrestrial plants, polyamines (small aliphatic amines) have been shown to be involved in protecting cells from damage under conditions of stress including UV-B radiation; such mechanisms have yet to be identified in algae. The overall aim of this study was to determine the importance of cellular processes in shaping the community structure of Porphyra on a wave-exposed shore on the east coast of the South Island, New Zealand. Porphyra distribution and community structure was assessed by regular monthly monitoring of presence and absence of Porphyra along four transect lines at the site. Enviromnental information was recorded to determine the effects of temperature, light, UV radiation, humidity and wind on Porphyra�s spatial and temporal distribution. Regular tissue samples were taken for species identification by the application of primers, which were specifically designed during this study. P. cinnamomea and Porphyra spec. "ROS 54" were identified as dominant species present almost throughout the year with a pronounced maximum in presence during late winter and spring, and some weeks of absence during April or May. The two dominant species were recorded from the low to the high intertidal shore, but the mid intertidal was identified as the preferred habitat. Other species that were found were rare and only present for a few months in a very restricted area. It was hypothesised that free radical generation and antioxidant metabolism are associated with desiccation tolerance in Porphyra. An attempt was made to investigate the impact of desiccation stress on Porphyra. The extraction process of antioxidants was problematic and no reproducible results could be obtained. It was attempted to investigate the spatial distribution of spores and conchocelis of different Porphyra species in the field, and determine if those found at Brighton Beach are species-specific in their morphology. This indicated that the two main Porphyra species at Brighton Beach not only prefer to occupy the same habitat but that they also have a morphologically similar conchocelis phase. Mechanisms on a cellular level such as polyamine metabolism affected by environmental (abiotic) stresses are related to the alga�s ability to adapt to stress and therefore can have an effect on Porphyra�s distribution along the shore and its presence throughout the year. The depletion of the ozone layer has become an important issue as the effects of increased UV radiation on the environment, especially the intertidal habitat, are revealed. Marine macrophytes possess the main three. polyamines: putrescine, spermidine and spermine of varying levels. For the few species studied, Rhodophyta generally contain higher levels of polyamines than Chlorophyta, while polyamine levels for the one heterokontophyte analysed were between Chlorophyta and Rhodophyta. Levels of the three most common polyamines (putrescine, spermidine, spermine) were determined in P. cinnamomea under controlled UV exposure. Tissue discs were exposed to visible light (PAR), PAR and UV-A or PAR, UV-A and UV-B radiation. Discs exposed to PAR and PAR and UV-A showed little change in polyamine levels over a six day trial period, while discs exposed to PAR, UV-A and UV-B showed a significant increase in free, bound soluble and bound insoluble polyamines over the same period of time. Correspondingly levels of ADC and ODC, two enzymes involved in polyamine synthesis, were measured. ODC levels changed little while ADC levels increased significantly during UV-B treatment, indicating that under UV-B stress polyamines are mainly synthesized via the ADC pathway. The experimental set-up and process of this study has not been applied in macroalgal polyamine research and results obtained are the first indication that increased levels of polyamines are involved in protection and/or protection mechanisms in macrophytic algae to prevent UV-B damage.

Magister Scientiae - MSc (Chemistry) / Plants have been widely used in traditional medicine for a number of ailments, among which may be included infectious diseases such as colds, influenza, chicken pox, TB, etc. as well as lifestyle diseases such as diabetes and cancer. Seaweeds have also been shown to contribute to the maintenance of health through their nutritional and medicinal properties and recently, a great deal of interest has developed towards the isolation of bioactive compounds from marine sources due to their numerous health benefits. Furthermore, marine algae are valuable sources of structurally diverse metabolites with scientifically proven therapeutic claims. Chemical constituents of red seaweed, Porphyra capensis was investigated in this present study along with subsequent brine shrimp lethality assay analysis of the crude extracts. The compounds isolated from the plant were from the hexane (6) and butanol (2) extracts. These compounds were all isolated and purified by various chromatographic techniques, namely silica gel chromatography, Sephadex LH-20 gel as well as C18 reversed phase silica gel. The structures of the isolated compounds were analysed and characterised by NMR, GC-MS, ESI MS and FTIR spectroscopy. Eight compounds were isolated and identified as phytol, desmosterol, 9-eicosenoic acid, 5,8,11,14,17-eicosapentanoic acid, palmitic acid, methyl (E)-hexadec-9-enoate, glycerol and compound 1 (novel compound). All the compounds were isolated from Porphyra capensis for the first time. The hexane, butanol and methanol extracts were found to be non-toxic with the brine shrimp test LC50 value at least two times greater than ?g/ml.