Causal networks of phytoplankton diversity and biomass are modulated by environmental context.

Untangling causal links and feedbacks among biodiversity, ecosystem functioning, and environmental factors is challenging due to their complex and context-dependent interactions (e.g., a nutrient-dependent relationship between diversity and biomass). Consequently, studies that only consider separable, unidirectional effects can produce divergent conclusions and equivocal ecological implications. To address this complexity, we use empirical dynamic modeling to assemble causal networks for 19 natural aquatic ecosystems (N24◦~N58◦) and quantified strengths of feedbacks among phytoplankton diversity, phytoplankton biomass, and environmental factors. Through a cross-system comparison, we identify macroecological patterns; in more diverse, oligotrophic ecosystems, biodiversity effects are more important than environmental effects (nutrients and temperature) as drivers of biomass. Furthermore, feedback strengths vary with productivity. In warm, productive systems, strong nitrate-mediated feedbacks usually prevail, whereas there are strong, phosphate-mediated feedbacks in cold, less productive systems. Our findings, based on recovered feedbacks, highlight the importance of a network view in future ecosystem management.

Long-term warming destabilizes aquatic ecosystems through weakening biodiversity-mediated causal networks.

Understanding how ecosystems will respond to climate changes requires unravelling the network of functional responses and feedbacks among biodiversity, physicochemical environments, and productivity. These ecosystem components not only change over time but also interact with each other. Therefore, investigation of individual relationships may give limited insights into their interdependencies and limit ability to predict future ecosystem states. We address this problem by analyzing long-term (16-39 years) time series data from 10 aquatic ecosystems and using convergent cross mapping (CCM) to quantify the causal networks linking phytoplankton species richness, biomass, and physicochemical factors. We determined that individual quantities (e.g., total species richness or nutrients) were not significant predictors of ecosystem stability (quantified as long-term fluctuation of phytoplankton biomass); rather, the integrated causal pathway in the ecosystem network, composed of the interactions among species richness, nutrient cycling, and phytoplankton biomass, was the best predictor of stability. Furthermore, systems that experienced stronger warming over time had both weakened causal interactions and larger fluctuations. Thus, rather than thinking in terms of separate factors, a more holistic network view, that causally links species richness and the other ecosystem components, is required to understand and predict climate impacts on the temporal stability of aquatic ecosystems.

Identification of a new mating group and reproductive isolation in the Closterium peracerosum-strigosum-littorale complex.

Reproductive isolation is essential for the process of speciation. In order to understand speciation, it is necessary to compare one mating group with other phylogenetically related but reproductively isolated groups. The Closterium peracerosum-strigosum-littorale (C. psl.) complex is a unicellular isogamous zygnematophycean alga, which is believed to share a close phylogenetic relationship with the land plants. In this study, we identified a new mating group, named group G, of C. psl. complex and compared its physiological and biochemical characteristics with the mating group I-E, which was closely related to the mating group G. Zygospores are typically formed as a result of conjugation between mating-type plus (mt+) and mating-type minus (mt-) cells in the same mating group during sexual reproduction. Crossing experiments revealed mating groups G and I-E were reproductively isolated from each other, but the release of lone protoplasts from mt- cells of mating group G was induced in the presence of mt+ cells of mating group I-E. In fact, the sex pheromone, protoplast-release-inducing protein of mating group I-E induced the release of protoplasts from mt- cells of mating group G. When mt+ and mt- cells of both mating groups I-E and G were co-cultured (multiple-choice matings), the zygospore formation of mating group G, but not that of mating group I-E, was inhibited. Based on these results, we propose a possible mechanism of reproductive isolation between the two mating groups and suggest the presence of sexual interference between mating group G and mating group I-E.

Morphology, taxonomy and mating-type loci in natural populations of Volvox carteri in Taiwan.

BACKGROUND: Volvox carteri f. nagariensis is a model taxon that has been studied extensively at the cellular and molecular level. The most distinctive morphological attribute of V. carteri f. nagariensis within V. carteri is the production of sexual male spheroids with only a 1:1 ratio of somatic cells to sperm packets or androgonidia (sperm packet initials). However, the morphology of male spheroids of V. carteri f. nagariensis has been examined only in Japanese strains. In addition, V. carteri f. nagariensis has heterothallic sexuality; male and female sexes are determined by the sex-determining chromosomal region or mating-type locus composed of a > 1 Mbp linear chromosome. Fifteen sex-specific genes and many sex-based divergent shared genes (gametologs) are present within this region. Thus far, such genes have not been identified in natural populations of this species. RESULTS: During a recent fieldwork in Taiwan, we encountered natural populations of V. carteri that had not previously been recorded from Taiwan. In total, 33 strains of this species were established from water samples collected in Northern Taiwan. Based on sequences of the internal transcribed spacer 2 region of nuclear ribosomal DNA and the presence of asexual spheroids with up to 16 gonidia, the species was clearly identified as V. carteri f. nagariensis. However, the sexual male spheroids of the Taiwanese strains generally exhibited a 1:1 to > 50:1 ratio of somatic cells to androgonidia. We also investigated the presence or absence of several sex-specific genes and the sex-based divergent genes MAT3m, MAT3f and LEU1Sm. We did not identify recombination or deletion of such genes between the male and female mating-type locus haplotypes in 32 of the 33 strains. In one putative female strain, the female-specific gene HMG1f was not amplified by genomic polymerase chain reaction. When sexually induced, apparently normal female sexual spheroids developed in this strain. CONCLUSIONS: Male spheroids are actually variable within V. carteri f. nagariensis. Therefore, the minimum ratio of somatic cells to androgonidia in male spheroids and the maximum number of gonidia in asexual spheroids may be diagnostic for V. carteri f. nagariensis. HMG1f may not be directly related to the formation of female spheroids in this taxon.

Phytoplankton dynamics of a subtropical reservoir controlled by the complex interplay among hydrological, abiotic, and biotic variables.

This study was conducted to identify the key factors related to the spatiotemporal variations in phytoplankton abundance in a subtropical reservoir from 2006 to 2010 and to assist in developing strategies for water quality management. Dynamic factor analysis (DFA), a dimension-reduction technique, was used to identify interactions between explanatory variables (i.e., environmental variables) and abundance (biovolume) of predominant phytoplankton classes. The optimal DFA model significantly described the dynamic changes in abundances of predominant phytoplankton groups (including dinoflagellates, diatoms, and green algae) at five monitoring sites. Water temperature, electrical conductivity, water level, nutrients (total phosphorus, NO3-N, and NH3-N), macro-zooplankton, and zooplankton were the key factors affecting the dynamics of aforementioned phytoplankton. Therefore, transformations of nutrients and reactions between water quality variables and aforementioned processes altered by hydrological conditions may also control the abundance dynamics of phytoplankton, which may represent common trends in the DFA model. The meandering shape of Shihmen Reservoir and its surrounding rivers caused a complex interplay between hydrological conditions and abiotic and biotic variables, resulting in phytoplankton abundance that could not be estimated using certain variables. Additional water quality and hydrological variables at surrounding rivers and monitoring plans should be executed a few days before and after reservoir operations and heavy storm, which would assist in developing site-specific preventive strategies to control phytoplankton abundance.

Taxonomic revision of Chlamydomonas subg. Amphichloris (Volvocales, Chlorophyceae), with resurrection of the genus Dangeardinia and descriptions of Ixipapillifera gen. nov. and Rhysamphichloris gen. nov.

Chlamydomonas (Cd.) is one of the largest but most polyphyletic genera of freshwater unicellular green algae. It consists of 400-600 morphological species and requires taxonomic revision. Toward reclassification, each morphologically defined classical subgenus (or subgroup) should be examined using culture strains. Chlamydomonas subg. Amphichloris is characterized by a central nucleus between two axial pyrenoids, however, the phylogenetic structure of this subgenus has yet to be examined using molecular data. Here, we examined 12 strains including six newly isolated strains, morphologically identified as Chlamydomonas subg. Amphichloris, using 18S rRNA gene phylogeny, light microscopy, and mitochondria fluorescent microscopy. Molecular phylogenetic analyses revealed three independent lineages of the subgenus, separated from the type species of Chlamydomonas, Cd. reinhardtii. These three lineages were further distinguished from each other by light and fluorescent microscopy-in particular by the morphology of the papillae, chloroplast surface, stigmata, and mitochondria-and are here assigned to three genera: Dangeardinia emend., Ixipapillifera gen. nov., and Rhysamphichloris gen. nov. Based on the molecular and morphological data, two to three species were recognized in each genus, including one new species, I. pauromitos. In addition, Cd. deasonii, which was previously assigned to subgroup "Pleiochloris," was included in the genus Ixipapillifera as I. deasonii comb. nov.

Integrating an algal ß-carotene hydroxylase gene into a designed carotenoid-biosynthesis pathway increases carotenoid production in yeast.

The algal ß-carotene hydroxylase gene Crchyb from Chlamydomonas reinhardtii, Czchyb from Chlorella zofingiensis, or Hpchyb from Haematococcus pluvialis and six other carotenoid-synthesis pathway genes were co-integrated into the genome of a yeast host. Each of these three algal genes showed a higher efficiency to convert ß-carotene to downstream carotenoids than the fungal genes from Phaffia rhodozyma. Furthermore, the strain with Hpchyb displayed a higher carotenoid productivity than the strains integrated with Crchyb or Czchyb, indicating that Hpchyb is more efficient than Crchyb and Czchyb. These results suggest that ß-carotene hydroxylase plays a crucial role in the biosynthesis of carotenoids.

Diatomological investigation in sphenoid sinus fluid and lung tissue from cases of suspected drowning.

We report on the presence, distribution and numbers of diatoms within specific organs as a result of drowning in fresh, treated and seawater. Specimens of sphenoid sinus fluid and lung tissue from 100 cases of suspected drowning and 20 cases where death was by natural causes, to act as a control, were examined for the presence of diatoms. In the 100 cases where the deceased was suspected to have drowned, 94 were confirmed as a death by drowning after autopsy with the other six being reported as death by another cause. No diatoms were found in cases confirmed as death by causes unrelated to drowning, with the exception of possible contamination via open wounds and through decomposition. In 94 cases, where all fatalities were confirmed as death by drowning, there were 81 cases in which diatoms were detected in samples taken from the sphenoid sinus fluid and/or lung tissue. No, or only few, diatoms were observed from the samples where the deceased drowned in treated waters such as spa or swimming pools. A significantly higher number of diatoms were detected in the sphenoid sinus fluid and lung tissue of confirmed drowning cases in fresh water compared to seawater. More diatoms were observed in sphenoid sinus fluid compared to lung tissue regardless of the water in which the deceased drowned. This study illustrates the potential use of diatom screening using both sphenoid sinus fluid and lung tissue to determine the cause of death in suspected cases of drowning. This report also highlights specific variables that need to be considered prior to such as conclusion being reached.

Linking secondary structure of individual size distribution with nonlinear size-trophic level relationship in food webs.

Existing individual size distribution (ISD) theories assume that the trophic level (TL) of an organism varies as a linear function of its log-transformed body size. This assumption predicts a power-law distribution of the ISD, i.e., a linear relationship between size and abundance in log space. However, the secondary structure of ISD (nonlinear dome shape structures deviating from a power-law distribution) is often observed. We propose a model that extends the metabolic theory to link the secondary structure of ISD to the nonlinear size-TL relationship. This model is tested with empirical data collected from a subtropical reservoir. The empirical ISD and size-TL relationships were constructed by FlowCAM imaging analysis and stable isotope analyses, respectively. Our results demonstrate that the secondary structure of ISD can be predicted from the nonlinear function of size-TL relationship and vice versa. Moreover, these secondary structures arise due to (1) zooplankton omnivory and (2) the trophic interactions within microbial food webs.

Tolerance of soil algae and cyanobacteria to drought stress.

Tolerance to drought stress in soil crust microorganisms is essential for exploiting suitable organisms for restoring soil. In this study, the responses to drought stress of two drought-tolerant species, a green alga and a cyanobacterium, were compared with those of two non-tolerant green algae. In response to drought stress, induced by treatment with polyethylene glycol, the intracellular proline levels increased and were associated with increases in malondialdehye, pigment contents, and enzyme activities such as superoxide dismutase (SOD) and peroxidase (POD). Our results suggest that tolerance to drought stress could be indicated by the intracellular levels of proline, SOD, and carotenoids. This study provides insights into the drought physiology of the photosynthetic microorganisms and suggests that Leptolyngbya boryana and Chlorella vulgaris are suitable pioneer organisms for soil restoration.

Biodiversity of soil algae in the farmlands of mid-Taiwan.

BACKGROUND: Very little information is available about soil algae in Taiwan. In this study, we investigated for the first time the soil algae inhabiting five types of farmland in mid-Taiwan: rice paddies, vegetable farms, tea plantations, sugar cane farms, and orchards. RESULTS: Sixty-four taxa belonging to 33 genera of cyanobacteria, diatoms, green algae, and euglenoids were identified on the basis of fine structures observed under optical and electron microscopes and rDNA sequencing. The majority of the isolates were of the genera Oscillatoria, Navicula, Nitzschia, and Pinnularia. Five species were reported for the first time in Taiwan, namely Microcoleus paludosus, M. subtorulosus, Navicula subminuscula, Nitzschia levidensis, and Ni. pusilla. CONCLUSIONS: The distribution and diversity of these species was shown to be strongly dependent on habitat, with the highest diversity of green algae in the orchards, a fairly high diversity of diatoms and cyanobacteria in the rice paddies, and a relatively low diversity in the tea plantations and sugar cane farms. The humidity and acidity were the most important environmental factors influencing the diversity of soil algae in these farmlands.

Production of phthalate esters by nuisance freshwater algae and cyanobacteria.

Phthalate esters are widely distributed pollutants which originate from synthetic plasticizer and are known to act as toxicants as well as environmental pheromones in the aquatic ecosystems. From investigating sixteen species of freshwater algae and cyanobacteria we revealed that some of them were capable of producing either di(n-butyl)phthalate (DBP) or mono(2-ethylhexyl)phthalate (MEHP) or both. These phthalate esters would be released into the environment under stress conditions. The incubation of the cells in culture medium containing NaH(13)CO(3) confirmed that both phthalates were de novo synthesized by the studied cells. This study suggested that the nuisance freshwater micro-algae and cyanobacteria growing in eutrophic waters might affect the aquatic ecosystem via the production of these phthalate esters.

Biochemical characterization of the small ubiquitin-like modifiers of Chlamydomonas reinhardtii.

Dynamic modification of target proteins by small ubiquitin-like modifier (SUMO) is known to modulate many important cellular processes and is required for cell viability and development in all eukaryotes. However, understanding of SUMO systems in plants, especially in unicellular green algae, remains elusive. In this study, Chlamydomonas reinhardtii CrSUMO96, CrSUMO97 and CrSUMO148 were characterized. We show that the formation of polymeric CrSUMO96 and CrSUMO97 chains can be catalyzed either by the human SAE1/SAE2 and Ubc9 SUMOylation system in vitro or by an Escherichia coli chimeric SUMOylation system in vivo. An exposed C-terminal di-glycine motif of CrSUMO96 or CrSUMO97 is essential for functional SUMOylation. The human SUMO-specific protease, SENP1, demonstrates more processing activity for CrSUMO97 than for CrSUMO96. The CrSUMO148 precursor notably has four repeated di-glycine motifs at the C-terminus. This unique feature is not found in other known SUMO proteins. Interestingly, only 83-residual CrSUMO148(1-83) with the first di-glycine motif can form SAE1/SAE2-SUMO complex and further form polymeric chains with the help of Ubc9. More surprisingly, CrSUMO148 precursor is digested by SENP1, solely at the peptide bond after the first di-glycine motif although there are four theoretically identical processing sites in the primary sequence. This process directly generates 83-residual CrSUMO148(1-83) mature protein, which is exactly the form suitable for activation and conjugation. We also show that SENP1 displays similar isopeptidase activity in the deconjugation of polymeric CrSUMO96, CrSUMO97 or CrSUMO148 chains, revealing that the catalytic mechanisms of processing and deconjugation of CrSUMOs by SENP1 may differ.

Effects of semidiurnal tidal circulation on the distribution of holo- and meroplankton in a subtropical estuary.

We examined how tidal changes and which physical factors affected holo- and meroplankton assemblages in a subtropical estuary in Taiwan in February 1999. A factor analysis showed that during tidal flooding, the water mass properties changed from low salinity (5-16) and high particulate organic carbon (POC, 2.6-4.5 mg L(-1)) content to increasing salinity and high total suspended matter content (29.0-104.5 mg L(-1)). With a receding tide, the water became more saline again, and its velocity increased (from non-detectable to 0.67 m s(-1)). One-way ANOVA showed that the distributions of four dominant taxa were affected by the ebb tide and exhibited two distinct groups. The first group consisted of non-motile invertebrate eggs and weakly swimming polychaete sabellid embryos and larvae (at densities of 1.25-1.40 ind. L(-1)), while the second consisted of better-swimming copepods and polychaete spionid larvae (at densities of 0.70-1.65 ind. L(-1)). A canonical correlation analysis demonstrated that the former group occurred at sites with greater freshwater input, higher POC content and greater depth, whereas the latter group was significantly associated with sites subject to seawater and faster flows. We propose that a two-layered circulation process and tidally induced oscillations in water movements might account for the distributional differences between these two groups.

PRODUCTION OF NATURAL BUTYLATED HYDROXYTOLUENE AS AN ANTIOXIDANT BY FRESHWATER PHYTOPLANKTON(1).

Butylated hydroxytoluene (BHT) is one of the synthetic antioxidant agents commonly used for food additives. In the present study, we determined that four freshwater phytoplankton, including a green alga (Botryococcus braunii Kütz.) and three cyanobacteria [Cylindrospermopsis raciborskii (Wollosz.) Seenaya et Sabba Raju, Microcystis aeruginosa (Kütz.) and Oscillatoria sp.] were capable of producing this compound. Hexane extracts from all the studied species exhibited various degrees of antioxidative properties when they were tested with the ß-carotene-linoleate (ß-CL) assay and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging assay. The highest antioxidant activity was observed in the crude extracts of M. aeruginosa and B. braunii, which displayed a similar activity to synthetic BHT. Gas chromatography/mass spectroscopy (GC-MS) analysis of the purified fractions revealed that the active compound was identical to synthetic BHT. Culturing under various irradiances gave rise to different magnitudes of BHT production in cyanobacterial cells, showing that more BHT was produced in the cells irradiated with a higher light intensity, and its production was irradiance dependent. Moreover, the quantity of cellular BHT displayed a positive correlation with the antioxidative activity of the tested species. The present study confirms the production of BHT in all four of the studied freshwater phytoplankton and suggests that these species constitute a potential source for producing natural BHT.

Simulation of water quality and plankton dynamics in the Danshuei River estuary, Taiwan.

An ecosystem model was developed to simulate the water quality and plankton dynamics in the Danshuei River estuary, Taiwan. The model simulates the hydrodynamics with a laterally integrated 2-dimensional intratidal numerical model, which supplies the physical transport processes for simulation of water quality and plankton state variables. The application of the model to the Danshuei River estuary indicates that the point source loadings are mainly responsible for the degraded water quality and very high nutrient concentrations in the estuary. The impacts of wastewater discharges are tightly controlled by the transport processes. Frequent occurrence of high river flow and flood events rapidly cleanses the estuary by flushing out both pollutants and plankton populations. The plankton is allowed to grow to significant populations if low river flow lasts for a period much longer than the biological time scale.

Can fog contribute to the nutrition of Chamaecyparis obtusa var. formosana? Uptake of a fog solute tracer into foliage and transport to roots.

Yellow cypress (Chamaecyparis obtusa (Siebold & Zucc.) Endl. var. formosana (Hayata) Rehder) is the predominant tree species of Taiwan's nutrient-poor, mountain fog forests. Little is known about the potential contribution of solute uptake from fog to the overall nutrition of these trees. Shoots of yellow cypress seedlings were misted with artificial fog containing the tracer rubidium (Rb) in laboratory and field experiments to determine if there is solute uptake from the fog. After misting shoots for six weeks, substantial amounts of tracer were detected in unexposed roots by inductively coupled plasma mass spectroscopy bulk analysis. Possible routes of entry were examined by element imaging with energy dispersive X-ray analysis. Direct uptake of the tracer into leaves across the cuticle and epidermis was small, excluding this as the major uptake path. Accumulations of Rb were found on leaf surfaces along the edges of the leaves. The almost daily changes in fog coverage and air humidity may enhance the accumulation of fog solutes at leaf edges. Accumulation of Rb was also found in narrow clefts between opposite leaves and between the outermost and underlying alternating stacked leaves. The clefts provide a direct passage from the leaf surface to the space beneath the imbricate leaves and the underlying alternate leaves, possibly facilitating solute uptake from fog, which in turn may contribute to the nutrition of yellow cypress.

Cytotoxic effects of free fatty acids on phytoplankton algae and cyanobacteria.

Some free fatty acids are toxic to phytoplankton, and the toxic effects are multiple. However, precisely how they kill phytoplankton is debatable. Here we show that fatty acids result in damage to plasma membranes, which might account for their lethal effects on phytoplankton. In this study, we used two chlorophytes (Chlorella vulgaris Beij. and Monoraphidium contortum (Thur.) Kom.-Legn.) and a cyanobacterium (Anabaena P-9) as test organisms. When these organisms were treated with deleterious concentrations of fatty acids, a remarkable elevation of extracellular potassium (K+) was detected in the culture medium; this indicates that leakage of intracellular K+ occurred as a result of damage to the plasma membranes. Exposure to unsaturated fatty acids resulted in higher levels of leaked K+ than did exposure to saturated ones, and levels of leakage displayed a positive correlation with the susceptibility of the growth of organisms to fatty acids. Stressed phytoplankton cells also exhibited cell lysis followed by free release of phycobilins. The sequence of cytotoxic effects elucidated here suggests that fatty acids primarily affect the plasma membranes, leading to a change in membrane permeability and dissociation of phycobilins from the thylakoids. Severe damage to the plasma membranes would give rise to a disruption of the stressed cells.

HPLC determination of pheophorbide a and pyropheophorbide a in dried laver product implicated in food poisoning.

In an outbreak of food poisoning involving a dried purple laver product (called nori), four persons had allergic-like symptoms such as inflammation and red rash on their face, mouth and belly. The causative nori was extracted and smeared on the arm-skin of five volunteers. Three out of five volunteers had a slight allergic reaction after 5 to 30 min when they were exposed to sunlight. The levels of the chlorophyll derivatives, pheophorbide a and pyropheophorbide a, measured by HPLC were 851-906 and 5,460-5,624 microg/g, respectively, in the causative samples. Judging from the high contents of pyropheophorbide a and pheophorbide a and the symptoms of patients and volunteers, the causative agents were concluded to be the photosensitizers pyropheophorbide a and pheophorbide a.