Abstract This research investigated the distribution of Trichodesmium in the South China Sea and the ecological factors that affect it. The survey periods were from August 1999 to October 2000. A total of 5 cruises were conducted, mostly in summer and autumn, including cruise 559 (August 18~25, 1999), cruise 629 (May 22~27, 2000), cruise 588 (July 3~14, 2000), cruise 597 (October 4-13, 2000), and cruise 657 (October 16-21, 2000). The sampling stations were located on the continental shelves, the slope zones, and the basin in the South China Sea between 13~22∘N and 114~120∘E. The density of Trichodesmium in the South China Sea varied greatly among the continental shelves, the slope zones, and the basin zones. Trichodesmium density in the continental shelf and the slope was high, with an average density of 150¡Ó297¡Ñ103 trichomes/m3 (4~1008¡Ñ103 trichomes/m3) in the surface water and 151¡Ó322¡Ñ103 trichomes/m3 (13~1114¡Ñ103 trichomes/m3) in 2-60m water depth. This could be caused by the influence of the mixing of the Kuroshio water and the substances of terrestrial origin. In the basin, Trichodesmium density was low, with an average density of 5¡Ó7¡Ñ103 trichomes/m3 (0~22¡Ñ103 trichomes/m3) in the surface water and an average density of 6¡Ó7¡Ñ103 trichomes/m3(0~22¡Ñ103 trichomes/m3) in 2~60m water depth. The Trichodesmium communities are mainly composed of T. thiebautii and T. erythraeum, with T. thiebautii being more abundant than T. erythraeum. Their relative importance at the continental shelf and the slope zones was 58% and 36% respectively, and 64% and 31% in the basin respectively. Trichodesmium appeared commonly as a single trichome, rarely in colony. The vertical distribution of Trichodesmium concentrated at 0-100m. The highest density was near the surface, varying between 4~1008¡Ñ103 trichomes/m3. In summer, the Trichodesmium in the continental shelf and the slope zone of the South China Sea was slightly higher than that in the East China Sea. The influences of temperature, nutrient abundance and wind speed on the distribution of Trichodesmium are not obvious. Trichodesmium in shallow water became abundant after the passage of a typhoon. The maximum density of Trichodesmium was observed in shallow water and average number of trichome per colony decreased. The influences of the typhoon on Trichodesmium species composition and colony density were not obvious. High density of Trichodesmium occurred at shallow waters and the water column with low stratification index (SI) which repressed the properties of waters in the continental shelf and the slope zones. The regions in which high-density Trichodesmium appeared, were the continental shelves and slope which showed water properties similar to those of the Kuroshio water, indicating possible intrusion of the Kuroshio water in summer into the continental shelves and slope of the northern South China Sea.

A fixação de N2 é uma fonte importante de nitrogênio novo nos ciclos biogeoquímicos, realizado potencialmente por cianobactérias diazotróficas, principalmente em condições de limitação de nitrogênio. O canal de São Sebastião (CSS) é reconhecidamente meso-oligotrófico, porém, eventos episódicos de precipitação podem alterar as concentrações de nutrientes locais interferindo na ocorrência de diazotróficas. Este trabalho investigou o efeito do aumento da pluviosidade na ocorrência e coexistência de diazotróficas no CSS e na baía do Araçá (BA), em 9 cenários distintos. As diazotróficas Trichodesmium spp. e Richelia intracellularis ocorreram após o aumento da pluviosidade nos dois pontos, não co-existindo em dois eventos. R. intracellularis ocorreu somente em endossimbiose com diatomáceas do gênero Hemiaulus, com predomínio em H. membranaceus. Os ventos de SW no CSS indicaram relações diretas na ocorrência das diazotróficas, sugerindo advecção de células provindas de outros locais. A temperatura e o fosfato apresentaram relações significativas com a ocorrência das diazotróficas. As densidades de diazotróficas e da comunidade fitoplanctônica foram maiores na BA em todos os eventos, assim como as concentrações de clorofila-a e nutrientes, ressaltando a importância da interação das águas da BA com o CSS. / The biological nitrogen fixation adds new nitrogen into the biogeochemical cycle. This process is attributed to diazothrophic cyanobacteria, which are known to occur in environments with nitrogen limitation. Although the São Sebastião Channel (CSS) is a meso-oligothrophic environment, local rain can affect nutrients concentrations, hence the occurrence of diazothrophic species. This work addressed how the increase in the precipitation rate alters the occurrence and co-existence of diazothrophic cyanobacteria in nine scenarios in the CSS and Araçá Bay (BA). The diazothrophic species Trichodesmium spp. and Richelia intracellularis occurred after an increase in rainfall, co-occurring in 7 of the 9 scenarios. Richelia intracellularis was always associated symbiotically with Hemiaulus diatoms, specially H. membranaceus. The occurrence of diazothrophic species was highly correlated to intense SW winds, suggesting the allochthonous input of cells in the CSS. The diazothrophic species occurrence was also associated with variations in temperature and phosphate concentration. The abundance of both diazothrophic species and phytoplankton community were higher in the BA in every scenario. The nutrients and chlorophyll-a concentrations were higher in the BA, highlighting the importance of water transport between BA and CSS.

A fixação de N2 é uma fonte importante de nitrogênio novo nos ciclos biogeoquímicos, realizado potencialmente por cianobactérias diazotróficas, principalmente em condições de limitação de nitrogênio. O canal de São Sebastião (CSS) é reconhecidamente meso-oligotrófico, porém, eventos episódicos de precipitação podem alterar as concentrações de nutrientes locais interferindo na ocorrência de diazotróficas. Este trabalho investigou o efeito do aumento da pluviosidade na ocorrência e coexistência de diazotróficas no CSS e na baía do Araçá (BA), em 9 cenários distintos. As diazotróficas Trichodesmium spp. e Richelia intracellularis ocorreram após o aumento da pluviosidade nos dois pontos, não co-existindo em dois eventos. R. intracellularis ocorreu somente em endossimbiose com diatomáceas do gênero Hemiaulus, com predomínio em H. membranaceus. Os ventos de SW no CSS indicaram relações diretas na ocorrência das diazotróficas, sugerindo advecção de células provindas de outros locais. A temperatura e o fosfato apresentaram relações significativas com a ocorrência das diazotróficas. As densidades de diazotróficas e da comunidade fitoplanctônica foram maiores na BA em todos os eventos, assim como as concentrações de clorofila-a e nutrientes, ressaltando a importância da interação das águas da BA com o CSS. / The biological nitrogen fixation adds new nitrogen into the biogeochemical cycle. This process is attributed to diazothrophic cyanobacteria, which are known to occur in environments with nitrogen limitation. Although the São Sebastião Channel (CSS) is a meso-oligothrophic environment, local rain can affect nutrients concentrations, hence the occurrence of diazothrophic species. This work addressed how the increase in the precipitation rate alters the occurrence and co-existence of diazothrophic cyanobacteria in nine scenarios in the CSS and Araçá Bay (BA). The diazothrophic species Trichodesmium spp. and Richelia intracellularis occurred after an increase in rainfall, co-occurring in 7 of the 9 scenarios. Richelia intracellularis was always associated symbiotically with Hemiaulus diatoms, specially H. membranaceus. The occurrence of diazothrophic species was highly correlated to intense SW winds, suggesting the allochthonous input of cells in the CSS. The diazothrophic species occurrence was also associated with variations in temperature and phosphate concentration. The abundance of both diazothrophic species and phytoplankton community were higher in the BA in every scenario. The nutrients and chlorophyll-a concentrations were higher in the BA, highlighting the importance of water transport between BA and CSS.

Thesis (Ph. D.)--Joint Program in Biological Oceanography (Massachusetts Institute of Technology, Dept. of Biology; and the Woods Hole Oceanographic Institution), 2009. / Cataloged from PDF version of thesis. / Includes bibliographical references (p. 149-162). / Trichodesmium is a colonial, N2-fixing cyanobacterium found in tropical oceans. Species of Trichodesmium are genetically similar but several species exist together in the same waters. In order to coexist, Trichodesmium spp. may occupy different niche spaces through differential utilization of resources such as nutrients and light, and through responses to physical characteristics such as temperature and turbulence. To investigate niche differentiation in Trichodesmium, I characterized cultured strains of Trichodesmium, identified and enumerated Trichodesmium clades in the field, and investigated P stress and N2 fixation in field populations. Species of Trichodesmium grouped into two clades based on sequences from 16S rDNA, the internal transcribed spacer (ITS), and the heterocyst differentiation gene hetR. Clade I contained Trichodesmium erythraeum and Trichodesmium contortum, and clade II contained Trichodesmium thiebautii, Trichodesmium tenue, Trichodesmium hildebrandtii, and Trichodesmium pelagicum. Each clade was morphologically diverse, but species within each clade had similar pigmentation. I developed a quantitative polymerase chain reaction (qPCR) method to distinguish between these two clades. In field populations of the Atlantic and Pacific Oceans, the qPCR method revealed that clade II Trichodesmium spp. were more prominent than clade I in the open ocean. Concentrations of Trichodesmium did not correlate with nutrient concentrations, but clade I had wider temperature and depth distributions than clade II. / (cont.) Temperature and light are physical characteristics that may define niche spaces for species of Trichodesmium. Clade I and II concentrations correlated with each other in the Pacific but not in the Atlantic, indicating that the two clades were limited by the same factors in the Pacific while different factors were limiting the abundance of the two clades in the Atlantic. Trichodesmium populations in the North Atlantic were more P stressed and had higher N2 fixation rates than populations in the western Pacific. While nutrient concentrations didn't directly correlate with Trichodesmium concentrations, the contrasting nutrient regimes found in the Atlantic and Pacific Oceans might influence distributions of the two clades differently. Unraveling the differences among species of Trichodesmium begins to explain their coexistence and enables us to understand factors controlling global N2 fixation. / by Annette Michelle Hynes. / Ph.D.

Nitrogen fixation is no longer considered to be a minor factor of the nitrogen cycle in oceanic ecosystems. Recent geochemical and biological efforts have led to a significant increase in the estimated input of nitrogen to marine ecosystems by biological fixation, while molecular studies have increased our knowledge of the number and diversity of nitrogen fixers known to be active in the ocean. Although Trichodesmium spp. have long been viewed as the primary marine nitrogen fixers, recent efforts have shown that various members of the picoplankton community are also actively involved in nitrogen fixation. The relative abundance of different nitrogen fixers is an important ecosystem parameter since nitrogen fixers may differ significantly in their physiology, life history and ecology. Here we combine rate measurements and stable isotope natural abundance measurements to constrain the impact of N2 fixation in the waters north of Australia. Samples were collected in the Coral, Arafura, and East Timor Seas, thus spanning three distinct hydrographic regions. Our data show that Trichodesmium has a significant influence on the stable nitrogen isotope ratios of particulate and zooplankton biomass and suggest that Trichodesmium is a significant source of nitrogen for the pelagic ecosystem. Based on stable carbon isotope ratios, it is also likely that the pathways are indirect and nitrogen fixed by Trichodesmium enters the higher trophic levels via decomposition as dissolved organic and inorganic nitrogen. Picocyanobacteria showed high diazotrophic activity at some stations, but unlike Trichodesmium, their N2 fixation rate was not reflected in the stable N isotope ratios of particulate and zooplankton biomass. Our results suggest an important N contribution to biomass by diazotrophs in the Coral Sea, Arafura Sea and East Timor Sea.

Zooplankton were sampled on a monthly basis at the Bermuda Atlantic Time-series Study (BATS) site from January 1995 to December 1999. Samples were collected using a 1-m², 200 µm mesh net. The net sampled the water column in an oblique manner from the surface to a mean depth of 200 m. One day and one night tow from each cruise was examined microscopically to determine the community structure of the non-calanoid copepods. In addition, a three year set of nighttime samples were examined taken by 0.5-m², 20 and 35 µm mesh nets (1995-1996 and 1997, respectively) towed obliquely to 150 m. The dominant orders in terms of overall abundance were the Cyclopoida and Poecilostomatoida. The cyclopoid genus, Oithona, was most abundant followed by the Poecilostomatoid family, Oncaeidae, and the genera Farranula and Corycaeus. Harpacticoids, although common, were about an order of magnitude less abundant and were dominated by Macrosetella gracilis. Representatives of the Mormonilloida and Siphonostomatoida also were frequently encountered, although in much lower numbers. Overall, pronounced seasonal signals were noted; highest abundances occurred during spring and lowest during winter. However, abundance of some groups peaked either in the fall or winter, with lowest abundance in spring or summer. Miraciid copepods are estimated to consume an overall average of 359 µg C m-2 d-1 and regenerate 55 µg N m-2 d-1 derived from Trichodesmium at BATS. Highest grazing and regeneration rates were found in late summer through fall and early winter and lowest in spring and early summer. The ecological consequences of miraciid copepod feeding on Trichodesmium are discussed. The 20-35 µm net samples revealed an astonishing abundance of non-copepod species, some totally missed and others woefully under-sampled by the 200 µm net. At least four species of oncaeid copepods and the harpacticoid copepod Microsetella norvegica were found in abundances that were more than an order of magnitude higher than the corresponding numbers of non-calanoid copepods sampled by the 200 µm net. The role of all non-calanoid copepods, from both net systems, in C and N dynamics at BATS is analyzed and discussed along with the sex-ratios of most identified species.

Iron is an essential nutrient for growth of photosynthetic microorganisms such as cyanobacteria and algae. Iron is required for proteins involved in the important processes of carbon and nitrogen assimilation. Low concentrations of iron in cultures or natural waters can lead to iron limitation which affects many aspects of algal metabolism. In natural waters, iron limitation can have effects on the patterns and rates of primary productivity. The cellular content of certain proteins can be affected by media iron concentrations. Methods have been used that assay components of the cell as an indirect measure of iron nutritional status. For example, spectroscopy can be performed to determine the cellular concentration of iron-containing proteins involved in photosynthesis. Organisms grown in media that imitate natural conditions, or organisms collected from their natural habitat are usually dilute. Methods that assay iron nutritional status such as spectroscopy and column chromatography require large sample sizes which are difficult to obtain from natural samples. In addition, methods that utilize techniques such as immunology or radioactive labelling are complex and time-consuming. These considerations led to the necessity of developing a technique that would be simple, rapid and effective on dilute samples. The method developed here utilized fast protein liquid chromatography (FPLC), which fulfilled these requirements. A complete analysis could be done within two to three hours with minimal sample treatment. The FPLC was simple to operate and was effective on a sample containing less than 100 μg of protein. Some photosynthetic organisms, when iron-depleted, can produce the flavin-containing protein flavodoxin (Flv). This protein substitutes for the iron-containing protein ferredoxin (Fd) in Fd-dependent reactions such as the light-induced reduction of NADP. The FPLC technique identified and quantified, in relative terms, Fd and Flv in the cell. Optical spectroscopy was used to verify FPLC retention time assignments. The results illustrated how the FPLC could be used to observe the changes in relative Fd and Flv content as a function of media iron concentration in cultures of the cyanobacterium Anabaena grown in the laboratory. It was found that Fd content decreased and Flv content increased with decreasing media iron concentration. In addition, samples of the cyanobacterium Trichodesmium collected from the ocean near Barbados were analyzed using FPLC to assay relative Fd and Flv content. By analogy with Anabaena, Fd and Flv retention times were identified. Using this technique conclusions could be drawn regarding the changing iron nutritional status of Trichodesmium in its natural habitat .