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• The relative N:P requirement for growth was explored for eight phytoplankton species. • Trophic status is negatively related to the N:P ratio required for optimal growth. • Both N and P are important for phytoplankton growth in eutrophic waters. • Phytoplankton tend to require higher N relative to P under low-nutrient conditions. • The results contribute to the ongoing debate of N vs. P limitation in freshwaters. Clarifying the pattern of relative nitrogen (N)-to-phosphorus (P) requirements for phytoplankton growth is of great significance for eutrophication mitigation and aquatic system management. The relative N-to-P requirement for phytoplankton growth is considered an essential trait determining species dominance within ecosystems and explaining phytoplankton response to nutrient availability. These requirements vary with environmental trophic status, though this variation remains unclear. Here, we evaluated the relative N-to-P requirements under different absolute nutrient levels using previous and current experimental data on eight phytoplankton species (three studied by us and five extrapolated from previous studies). Results showed that relative N-to-P requirements for phytoplankton growth decreased as absolute nutrient levels increased. Thus, N may be crucial for enhancing phytoplankton growth under low nutrient conditions, whereas P may be the primary limiting factor of phytoplankton growth under sufficient nutrient conditions. This result applies to single species as well as species assemblages, which are independent of species shifts occurring along water N:P gradients. The response observed in our large trophic status gradient may help elucidate the relative importance of N and P reductions in mitigating the impact of eutrophication on ecosystems. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract: Members of the cyanobacterial genus Microcystis commonly form blooms in eutrophic freshwater systems, and some produce cyclic heptapeptide hepatotoxins called microcystins, thereby often causing serious water management problems. Microcystis species were unified into the single Microcystis aeruginosa classification based on 16S rRNA gene sequences and DNA–DNA re-association experiments; however, the morphological features of the organisms differ in different culturing conditions. Here, we describe a new real-time quantitative PCR (qPCR) method of determining Microcystis intradiversity using the SYBR Green I assay. We analyzed 71 Microcystis 16S-23S rDNA internal transcribed spacer region (16S-23S ITS) sequences, designed three group-specific PCR primers that successfully selected a morphologically M. wesenbergii-like non-toxic group (Group-3), and differentiated between M. viridis-like toxic group (Group-4) and M. aeruginosa-like Group-1 organisms including toxic and non-toxic Microcystis strains. The primers covered 76% of the Microcystis 16S-23S ITS regions from all over the world (six continents) included in GenBank. We constructed a mixed culture with representative Microcystis strains from each group, and estimated their cell densities by qPCR over 7 weeks. Group-1 and Group-3 grew exponentially for 4 weeks; however, the growth of Group-4 declined after 2 weeks, revealing different growth properties for the Microcystis groups in the mixed culture. Finally, we applied this method to natural Microcystis blooms at four freshwater sites, and found the dominance of Group-1 in three blooms and of Group-3 in one bloom, thereby showing the geographically uneven distribution of Microcystis genotypes. The developed qPCR technique targeting the 16S-23S ITS region is both rapid and simple and is useful for selective quantification of group variations among sympatric Microcystis genotypes, such as in mixed cultures and the natural environment. [Copyright &y& Elsevier]

Abstract: The size and composition of soil microbial communities have important influences on terrestrial ecosystem processes such as soil decomposition. However, compared with studies of aboveground plant communities, there are relatively few studies on belowground microbial communities and their interactions with aboveground vegetations in the arctic region. In this study, we conducted the first investigation of the abundance and composition of prokaryotic communities along small-scale vegetation gradients (ca. 1–3 m) in a dry arctic tundra ecosystem in Northern Sweden using fluorescent in situ hybridization (FISH) coupled with catalyzed reporter deposition (CARD). The number of prokaryotic cells increased with increasing vegetation cover along vegetation gradients, mainly as a function of increased amounts of soil carbon and moisture. Eubacteria and Archaea constituted approximately 59.7% and 33.4% of DAPI-positive cells, respectively. Among the analyzed bacterial phyla and sub-phyla, Acidobacteria and α-proteobacteria were the most dominant groups, constituting approximately 13.5% and 10.7% of DAPI-positive cells, respectively. Interestingly, the soil prokaryotic community composition was relatively unaffected by the dramatic changes in the aboveground vegetation community. Multivariate analyses suggested that the prokaryotic community composition depended on soil pH rather than on aboveground vegetation. Surface plants are weak predictors of the composition of the soil microbial community in the studied soil system and the size of the community is constrained by carbon and water availability. In addition, our study demonstrated that CARD-FISH, which is still a rarely-used technique in soil ecology, is effective for quantifying soil microbes. [Copyright &y& Elsevier]

Abstract: In this study, we sought to clarify the phylogenetic and ecological relationships of anatoxin-a-producing strains of Cuspidothrix issatschenkoi (Usačev) Rajaniemi et al. and Raphidiopsis mediterranea Skuja var. grandis Hill in Japanese lakes. We isolated 60 strains from six water bodies and evaluated their ability to produce anatoxin-a and nitrogenase, and determined the sequences of the 16S–23S rDNA internal transcribed spacer region. The results consistently indicated that all R. mediterranea ver. grandis-like strains were non-heterocytous, anatoxin-a-producing C. issatschenkoi. We also evaluated the distribution of anatoxin-a-producing strains of C. issatschenkoi in 88 water bodies using primers and a probe specifically developed in this study for quantitative PCR. By this method, we detected anatoxin-a-producing strains in 16 water bodies, while by automated rRNA intergenic spacer analysis, C. issatschenkoi was detected in 24 water bodies, indicating that the toxic and non-toxic strains occur in different environments. Although C. issatschenkoi has been classified as the functional group H1, which is tolerant to low nitrogen concentrations, the anatoxin-a-producing strains were detected from water bodies with low TOC/TON because of the lack of nitrogen-fixing ability. These results suggest that different genotypes of C. issatschenkoi have different ecotypes and prefer different environments. [Copyright &y& Elsevier]

Abstract: We studied the frequency and composition of potentially toxic cyanobacteria in 30 samples from 25 Japanese lakes using automated rRNA intergenic spacer analysis (ARISA) and eight primer sets for nodularin, microcystin, cylindrospermopsin, anatoxin-a, and saxitoxin synthetase genes. Potential microcystin- and anatoxin-a-producers were detected in 25 and 7 samples, respectively. Cylindrospermopsin- or saxitoxin-producers were not detected. PCR and clone library analyses indicated that Microcystis was the sole microcystin-producing genus. Moreover, potential microcystin-producing Microcystis strains were detected in 25 of 26 samples which included Microcystis ARISA fragments, suggesting that toxic Microcystis is ubiquitous. Potential anatoxin-a-producers detected in the samples were estimated to be Cuspidothrix issatschenkoi (synonym: Aphanizomenon issatschenkoi) from clone library analyses of the anatoxin-a biosynthesis gene cluster and 16S–23S rDNA intergenic spacer region. Anatoxin-a-producing strains of C. issatschenkoi are known from two lakes in New Zealand and Germany. In contrast, C. issatschenkoi was the second most common toxic cyanobacteria in our study. This is the first report of potential anatoxin-a-producing C. issatschenkoi strains in Asia. [Copyright &y& Elsevier]

Abstract: Management of the food web is important in maintaining ecosystem functions; however, the studies to assess anthropogenic impact on food web structures are very limited. Recently, shoreline development is considered to impact the littoral zone of lakes through loss of littoral habitats. Shoreline engineering would consequently modify the food web structure and coupling between benthic and pelagic habitats. Thus, we investigated whether lakeshore modification would change benthic–pelagic coupling by largemouth bass in 15 ponds, which differed in the proportion of concrete bank modification. We performed isotope mixing models to estimate benthic contribution to the fish species and tested the relationships with environmental factors including proportion of bank modification by general linear models. These results showed that the benthic contribution to largemouth bass was strongly related to the proportion of anthropogenic shoreline modification, while for bluegill sunfish, the benthic contribution was not changed with anthropogenic shoreline modification. Therefore, anthropogenic modification of shoreline strongly modified the food webs in ponds through the littoral habitat modification. The results suggested that we should pay attention to the anthropogenic impact on food web structures to maintain ecosystems. [Copyright &y& Elsevier]