Characterization of an Aerosol-Based Photobioreactor for Cultivation of Phototrophic Biofilms

Phototrophic biofilms, in particular terrestrial cyanobacteria, offer a variety of biotechnologically interesting products such as natural dyes, antibiotics or dietary supplements. However, phototrophic biofilms are difficult to cultivate in submerged bioreactors. A new generation of biofilm photobioreactors imitates the natural habitat resulting in higher productivity. In this work, an aerosol-based photobioreactor is presented that was characterized for the cultivation of phototrophic biofilms. Experiments and simulation of aerosol distribution showed a uniform aerosol supply to biofilms. Compared to previous prototypes, the growth of the terrestrial cyanobacterium Nostoc sp. could be almost tripled. Different surfaces for biofilm growth were investigated regarding hydrophobicity, contact angle, light- and temperature distribution. Further, the results were successfully simulated. Finally, the growth of Nostoc sp. was investigated on different surfaces and the biofilm thickness was measured noninvasively using optical coherence tomography. It could be shown that the cultivation surface had no influence on biomass production, but did affect biofilm thickness.

The two-component response regulator OrrA confers dehydration tolerance by regulating anaKa expression in the cyanobacterium Anabaena sp. strain PCC 7120

The aquatic cyanobacterium Anabaena sp. strain PCC 7120 exhibits dehydration tolerance. The regulation of gene expression in response to dehydration is crucial for the acquisition of dehydration tolerance, but the molecular mechanisms underlying dehydration responses remain unknown. In this study, the functions of the response regulator OrrA in the regulation of salt and dehydration responses were investigated. Disruption of orrA abolished or diminished the induction of hundreds of genes in response to salt stress and dehydration. Thus, OrrA is a principal regulator of both stress responses. In particular, OrrA plays a crucial role in dehydration tolerance because an orrA disruptant completely lost the ability to regrow after dehydration. Moreover, in the OrrA regulon, anaKa encoding a protein of unknown function was revealed to be indispensable for dehydration tolerance. OrrA and AnaK are conserved among the terrestrial cyanobacteria, suggesting their conserved functions in dehydration tolerance in cyanobacteria.

Metagenome Sequencing to Explore Phylogenomics of Terrestrial Cyanobacteria

Cyanobacteria are ubiquitous microorganisms with crucial ecosystem functions, yet most knowledge of their biology is from aquatic taxa. We have constructed metagenomes for 50 taxonomically well-characterized terrestrial cyanobacterial cultures. These data will support phylogenomic studies of evolutionary relationships and gene content among these unique algae and their aquatic relatives.

Geographical Variability of Mineral Elements and Stability of Restrictive Mineral Elements in Terrestrial Cyanobacteria Across Gradients of Climate, Soil, and Atmospheric Wet Deposition Mineral Concentration

Terrestrial cyanobacteria Nostoc commune is an ideal species to study the geographical variation of mineral elements of soil cyanobacteria at the species level. Here, we first address the following questions: (1) from where are these mineral elements, (2) are there geographical variations for these mineral elements, and if so, (3) which environmental factors drive the geographical variation of these mineral elements? Second, we tested whether the soil cyanobacterial mineral elements followed the “restrictive element stability hypothesis” of higher plants. Finally, we explored the effect of mineral geographic variation on ecological adaptation of soil cyanobacteria. We collected N. commune samples across gradients of climate, soil, and atmospheric wet deposition mineral concentration in mainland China. We measured fifteen minerals, including five macroelements (N, Ca, K, Fe, P), five microelements (Mn, Zn, Cu, Co, Se), and five heavy metals (Pb, Cr, As, Cd, Hg). We found that five elements (P, Cu, Zn, Co, Pb) had significant geographical variation. They increased as the distance from the equator increased and decreased as the distance from the prime meridian increased. Mean annual precipitation and mean annual temperature explained most of the variation. We did not find any significant correlations between the mineral element contents in N. commune and the minerals in soil and rainfall, except for P. There was no significant correlation between the variation coefficients of different elements and their actual detected contents and their potential physiological required contents. The statistical results of our experiment did not support the “restrictive element stability hypothesis.” We speculated that net accumulation of mineral elements in cyanobacterial cells and extracellular polysaccharides (EPS) might play an important role for terrestrial cyanobacteria in the adaptation to dry and cold conditions.

Materials to cyanobacterial and algal flora from volcanic soils of Shiveluch volcano

During the species composition study of terrestrial cyanobacteria and algae from volcanic soils of Shiveluch Volcano (Kamchatka peninsula, Russia) eighteen taxa from five phyla were revealed: Cyanobacteria – 4, Bacillariophyta – 4, Ochrophyta – 2 (Eustigmatophyceae – 1, Xanthophyceae – 1), Charophyta – 1, Chlorophyta – 7 (Chlorophyceae – 2, Trebouxiophyceae – 5). Nitzschia communis Rabenhorst, Nitzschia palea (Kützing) W. Smith, Eolimna minima (Grunow) Lange-Bertalot, Eremochloris sp., Tetradesmus obliquus (Turpin) M.J. Wynne, Nostoc edaphicum Kondratyeva were most frequency.

A qPCR method for distinguishing biomass from non-axenic terrestrial cyanobacteria cultures in hetero- or mixotrophic cultivations

The cultivation of cyanobacteria with the addition of an organic carbon source (meaning as heterotrophic or mixotrophic cultivation) is a promising technique to increase their slow growth rate. However, most cyanobacteria cultures are infected by non-separable heterotrophic bacteria. While their contribution to the biomass is rather insignificant in a phototrophic cultivation, problems may arise in heterotrophic and mixotrophic mode. Heterotrophic bacteria can potentially utilize carbohydrates quickly, thus preventing any benefit for the cyanobacteria. In order to estimate the advantage of the supplementation of a carbon source, it is essential to quantify the proportion of cyanobacteria and heterotrophic bacteria in the resulting biomass. In this work, the use of quantitative polymerase chain reaction (qPCR) is proposed. To prepare the samples, a DNA extraction method for cyanobacteria was improved to provide reproducible and robust results for the group of terrestrial cyanobacteria. Two pairs of primers were used, which bind either to the 16S rRNA gene of all cyanobacteria or all bacteria including cyanobacteria. This allows a determination of the proportion of cyanobacteria in the biomass. The method was established with the two terrestrial cyanobacteria Trichocoleus sociatus SAG 26.92 and Nostoc muscorum SAG B-1453-12a. As proof of concept, a heterotrophic cultivation with T. sociatus with glucose was performed. After 2 days of cultivation, a reduction of the biomass partition of the cyanobacterium to 90% was detected. Afterwards, the proportion increased again.