Characterization of polyphosphate dynamics in the widespread freshwater diatom Achnanthidium minutissimum under varying phosphorus supplies.

Polyphosphates (polyP) are ubiquitous biomolecules that play a multitude of physiological roles in many cells. We have studied the presence and role of polyP in a unicellular alga, the freshwater diatom Achnanthidium minutissimum. This diatom stores up to 2.0 pg·cell-1 of polyP, with chain lengths ranging from 130 to 500 inorganic phosphate units (Pi). We applied energy dispersive X-ray spectroscopy, Raman/fluorescence microscopy, and biochemical assays to localize and characterize the intracellular polyP granules that were present in large apical vacuoles. We investigated the fate of polyP in axenic A. minutissimum cells grown under phosphorus (P), replete (P(+)), or P deplete (P(-)) cultivation conditions and observed that in the absence of exogenous P, A. minutissimum rapidly utilizes their internal polyP reserves, maintaining their intrinsic growth rates for up to 8 days. PolyP-depleted A. minutissimum cells rapidly took up exogenous P a few hours after Pi resupply and generated polyP three times faster than cells that were not initially subjected to P limitation. Accordingly, we propose that A. minutissimum deploys a succession of acclimation strategies regarding polyP dynamics where the production or consumption of polyP plays a central role in the homeostasis of the diatom.

A semi-automated, KNIME-based workflow for biofilm assays.

BACKGROUND: A current focus of biofilm research is the chemical interaction between microorganisms within the biofilms. Prerequisites for this research are bioassay systems which integrate reliable tools for the planning of experiments with robot-assisted measurements and with rapid data processing. Here, data structures that are both human- and machine readable may be particularly useful. RESULTS: In this report, we present several simplification and robotisation options for an assay of bacteria-induced biofilm formation by the freshwater diatom Achnanthidium minutissimum. We also tested several proof-of-concept robotisation methods for pipetting, as well as for measuring the biofilm absorbance directly in the multi-well plates. Furthermore, we exemplify the implementation of an improved data processing workflow for this assay using the Konstanz Information Miner (KNIME), a free and open source data analysis environment. The workflow integrates experiment planning files and absorbance read-out data, towards their automated processing for analysis. CONCLUSIONS: Our workflow lead to a substantial reduction of the measurement and data processing workload, while still reproducing previously obtained results in the A. minutissimum biofilm assay. The methods, scripts and files we designed are described here, offering adaptable options for other medium-throughput biofilm screenings.

Biofilm and capsule formation of the diatom Achnanthidium minutissimum are affected by a bacterium.

Photoautotrophic biofilms play an important role in various aquatic habitats and are composed of prokaryotic and/or eukaryotic organisms embedded in extracellular polymeric substances (EPS). We have isolated diatoms as well as bacteria from freshwater biofilms to study organismal interactions between representative isolates. We found that bacteria have a strong impact on the biofilm formation of the pennate diatom Achnanthidium minutissimum. This alga produces extracellular capsules of insoluble EPS, mostly carbohydrates (CHO), only in the presence of bacteria (xenic culture). The EPS themselves also have a strong impact on the aggregation and attachment of the algae. In the absence of bacteria (axenic culture), A. minutissimum did not form capsules and the cells grew completely suspended. Fractionation and quantification of CHO revealed that the diatom in axenic culture produces large amounts of soluble CHO, whereas in the xenic culture mainly insoluble CHO were detected. For investigation of biofilm formation by A. minutissimum, a bioassay was established using a diatom satellite Bacteroidetes bacterium that had been shown to induce capsule formation of A. minutissimum. Interestingly, capsule and biofilm induction can be achieved by addition of bacterial spent medium, indicating that soluble hydrophobic molecules produced by the bacterium may mediate the diatom/bacteria interaction. With the designed bioassay, a reliable tool is now available to study the chemical interactions between diatoms and bacteria with consequences for biofilm formation.

Achnanthidiumgladius sp. nov. (Bacillariophyceae) - a new monoraphid diatom species from Indonesia.

A new monoraphid diatom species Achnanthidiumgladius sp. nov. is described from Indonesia. The description is based on molecular data (18SV4), morphological analysis and comparison with similar species. According to molecular data, Achnanthidiumgladius sp. nov. is closely related to Achnanthidiumminutissimum. Morphologically, the new species differs from similar species by the absence of a fascia on raphe valve, cell size, and striae density and pattern. The new species is only known from the type locality in Indonesia. Comparison with close related species is given.

Achnanthidium tinea sp. nov. - a new monoraphid diatom (Bacillariophyceae) species, described on the basis of molecular and morphological approaches.

A new monoraphid diatom species Achnanthidium tinea Tseplik, Kulikovskiy, Kociolek & Maltsev, sp. nov. is described from Indonesia. The species is described on the basis of molecular and morphological analyses. According to molecular data the new species belongs to the clade that includes strains of Achnanthidium minutissimum, Achnanthidium saprophilum and Achnanthidium digitatum. Morphologically, the new species differs quite significantly from other species of the same genus because of linear-elliptic valves with almost parallel sides and strongly radiate striae and a butterfly-shaped fascia on the raphe valve. The morphology and phylogeny of the new species are discussed, and thoughts on the current state of the taxonomy of the genus Achnanthidium are expressed. Our work shows the importance of using molecular data in diatom systematics and also demonstrates the need to investigate rarely studied regions of our planet.

[Community of Benthic Diatoms and Their Relationship with Aquatic Environmental Factors in the Tangwang River, China].

To reasonably evaluate the eco-environmental health of the Tangwang River, which is a tributary of the Songhua River in China, community structures of periphyton and cleanliness of the benthic diatom at 24 sampling sites were investigated using McNaughton's dominance index, clustering, and ecotype analysis, while the relationship between the environmental factors and the diatom communities were studied by principal component analysis, Spearman correlation test, and redundancy analysis, in August 2018 (flood season). A total of 99 species or variants of benthic diatoms have been identified, indicating that there were abundant diatoms in the Tangwang River. Achnanthidium minutissimum and other diatoms that can be used as clean water indicators were dominant species in the Tangwang River, which indicates that the eco-environmental quality of the Tangwang River was relatively healthy in the flood season. Of these, the dominant degree of A. minutissimum was 0.32, making it the absolute dominant species in Tangwang River. Sampling sites can be divided into three groups based on clustering analysis. The dominant species of group 1 and group 2 were mainly clean species, indicating that the two groups were in a relatively healthy state. Nitzschia palea, Ulnaria ulna, and other diatoms that can be used as eutrophication indicators were the dominant species of group 3, indicating that group 3 was less healthy than the other two groups. From groups 1 and 2, the results from ecotype analysis showed a decrease in the proportion of polyoxybiontic diatoms and an increase in the proportion of α-mesosaphrobe diatoms, polysaprobe diatoms, oligo-mesotrophic diatoms, mesotrophic diatoms, meso-eutrophic diatoms, and eutrophic diatoms. Compared to the other two groups, the results from ecotype analysis showed a significant increase in the proportion of α-mesosaphrobe diatoms, polysaprobe diatoms, eutrophic diatoms and hypereutrophic diatoms in group 3. The predominant aquatic influencing factors of diatom community structures for the Tangwang River were permanganate index, total nitrogen (TN), and ammonia nitrogen (NH4+-N), of which permanganate index was the main factor for group 2, while TN and NH4+-N were the main factors for group 3. As a result, the eco-environmental quality of the Tangwang River was good, and the benthic diatom was found to be an effective indicator of the nutritional conditions and saprophytic status.