Oxidative stress and starvation in Dinoroseobacter shibae: the role of extrachromosomal elements.

Aerobic anoxygenic phototrophic bacteria (AAP) are abundant in the photic zone of the marine environment. Dinoroseobacter shibae, a representative of the Roseobacter group, converts light into additional energy that enhances its survival especially under starvation. However, light exposure results in the production of cytotoxic reactive oxygen species in AAPs. Here we investigated the response of D. shibae to starvation and oxidative stress, focusing on the role of extrachromosomal elements (ECRs). D. shibae possessing five ECRs (three plasmids and two chromids) was starved for 4 weeks either in the dark or under light/dark cycles and the survival was monitored. Transcriptomics showed that on the chromosome genes with a role in oxidative stress response and photosynthesis were differentially expressed during the light period. Most extrachromosomal genes in contrast showed a general loss of transcriptional activity, especially in dark-starved cells. The observed decrease of gene expression was not due to plasmid loss, as all five ECRs were maintained in the cells. Interestingly, the genes on the 72-kb chromid were the least downregulated, and one region with genes of the oxygen stress response and a light-dependent protochlorophyllide reductase of cyanobacterial origin was strongly activated under the light/dark cycle. A Δ72-kb curing mutant lost the ability to survive under starvation in a light/dark cycle demonstrating the essential role of this chromid for adaptation to starvation and oxidative stress. Our data moreover suggest that the other four ECRs of D. shibae have no vital function under the investigated conditions and therefore were transcriptionally silenced.

Light enhances survival of Dinoroseobacter shibae during long-term starvation.

Aerobic anoxygenic phototrophs (AAPs) as being photoheterotrophs require organic substrates for growth and use light as a supplementary energy source under oxic conditions. We hypothesized that AAPs benefit from light particularly under carbon and electron donor limitation. The effect of light was determined in long-term starvation experiments with Dinoroseobacter shibae DFL 12(T) in both complex marine broth and defined minimal medium with succinate as the sole carbon source. The cells were starved over six months under three conditions: continuous darkness (DD), continuous light (LL), and light/dark cycle (LD, 12 h/12 h, 12 µmol photons m(-2) s(-1)). LD starvation at low light intensity resulted in 10-fold higher total cell and viable counts, and higher bacteriochlorophyll a and polyhydroxyalkanoate contents. This coincided with better physiological fitness as determined by respiration rates, proton translocation and ATP concentrations. In contrast, LD starvation at high light intensity (>22 µmol photons m(-2) s(-1), LD conditions) resulted in decreasing cell survival rates but increasing carotenoid concentrations, indicating a photo-protective response. Cells grown in complex medium survived longer starvation (more than 20 weeks) than those grown in minimal medium. Our experiments show that D. shibae benefits from the light and dark cycle, particularly during starvation.

Microbial community diversity of organically rich cassava sago factory waste waters and their ability to use nitrate and N2O added as external N-sources for enhancing biomethanation and the purification efficiency.

Water shortage necessitated South Indian sago factory owners, extracting starch out of cassava tubers, to install biogas plants where a starch utilizing microbial community multiplies and reduces the biological oxygen demand (BOD) of the waste waters by presently about 30%. The purification efficiency of sago factory waste waters, rich in solid particles and having wide C/N ratios, around 250, through unstirred biogas plants needs to be improved. Our approach was to apply instead of animal slurry nitrate (NO3(-)) and nitrous oxide (N2O) as external N-sources anticipating a better N-distribution in the unstirred biogas plants. Estimated cell numbers, bacterial community changes, on the basis of 16S rRNA gene clone libraries and changing CO2-, CH4-, N2O releases due to the presence of nitrate or N2O suggest that acid tolerant Lactobacillus spp. dominate the biogas plant inflows (pH 3.5). They were very less or not found in the outflows (pH 7.3). Assumingly, the phyla Bacteroidetes (Prevotella spp.), Proteobacteria (Rhizobium spp., Defluvibacter sp.), Firmicutes (Megasphaera spp., Dialister spp., Clostridium spp.) and Synergistetes (Thermanaerovibrio spp.), not-detectable in the biogas plant inflows, replaced them. Anaerobes, about 400cellsml(-1) in the inflows, increased to about 10(6)cellsml(-1) in the outflows. The methane formation, as confirmed by the incubation experiments, suggests that methanogens must have been present among the anaerobes. In the biogas plant in- and outflows also about 300cellsml(-1) denitrifying bacteria and up to 10(4)cfu fungi were found. Despite the low number of denitrifying bacteria nitrate added to the biogas plant in- and outflows was widely consumed and added N2O decreased considerably. Thus, wide C/N ratios substrates like sago factory waste waters keep the N2O emissions low by using N2O either as electron acceptor or by incorporating it into the growing biomass what needs to be confirmed. The biogas plant inflow samples have emitted tentatively more CO2 and the outflow samples released more CH4.