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Tuberculosis (TB) claims more human lives than any other infectious organism. The lethal synergy between TB-HIV infection and the rapid emergence of drug resistant strains has created a global public health threat that requires urgent attention. Mycobacterium tuberculosis, the causative agent of TB is an exquisitely well-adapted human pathogen, displaying the ability to promptly remodel metabolism when encountering stressful environments during pathogenesis. A careful study of the mechanisms that enable this adaptation will enhance the understanding of key aspects related to the microbiology of TB disease. However, these efforts require microbiological model systems that mimic host conditions in the laboratory. Herein, we describe several in vitro model systems that generate non-replicating and differentially culturable mycobacteria. The changes that occur in the metabolism of M. tuberculosis in some of these models and how these relate to those reported for human TB disease are discussed. We describe mechanisms that tubercle bacteria use to resuscitate from these non-replicating conditions, together with phenotypic heterogeneity in terms of culturabiliy of M. tuberculosis in sputum. Transcriptional changes in M. tuberculosis that allow for adaptation of the organism to the lung environment are also summarized. Finally, given the emerging importance of the microbiome in various infectious diseases, we provide a description of how the lung and gut microbiome affect susceptibility to TB infection and response to treatment. Consideration of these collective aspects will enhance the understanding of basic metabolism, physiology, drug tolerance and persistence in M. tuberculosis to enable development of new therapeutic interventions. [ABSTRACT FROM AUTHOR]

The decision to initiate DNA replication is a critical step in the cell cycle of all organisms. In nearly all bacteria, replication initiation requires the activity of the conserved replication initiation protein DnaA. Due to its central role in cell cycle progression, DnaA activity must be precisely regulated. This review summarizes the current state of DnaA regulation in the asymmetrically dividing α-proteobacterium Caulobacter crescentus , an important model for bacterial cell cycle studies. Mechanisms will be discussed that regulate DnaA activity and abundance under optimal conditions and in coordination with the asymmetric Caulobacter cell cycle. Furthermore, we highlight recent findings of how regulated DnaA synthesis and degradation collaborate to adjust DnaA abundance under stress conditions. The mechanisms described provide important examples of how DNA replication is regulated in an α-proteobacterium and thus represent an important starting point for the study of DNA replication in many other bacteria. This article is part of a Special Issue entitled: Dynamic gene expression, edited by Prof. Patrick Viollier. [ABSTRACT FROM AUTHOR]

The new species Parietochloris grandis sp. nov. is described from forest soil in the Dnipropetrovsk region, Ukraine. The description is based on morphological features and the phylogenetic analysis of partial SSU rDNA and rbcL genes. Phylogenetic analysis places P . grandis in the Parietochloris clade, within the family Trebouxiophyceae. The novel strain formed a strongly supported monophyletic lineage with the type species of Parietochloris , P . alveolaris . P . grandis differed from other species in the Parietochloris clade by the size and form of vegetative cells and the large number of zoospores in zoosporangia. A number of experiments with different phosphates and nitrates concentrations were conducted to evaluate changes in fatty acid profile and biomass. The dominant fatty acids during cultivation on standard BG-11 medium, as well as with the phosphates concentrations ranged from 0.22 to 2 mM, were linoleic acid (24–25%), palmitic acid (12–14%), linolenic acid (9–12%), and oleic acid (7–11%). The content of arachidonic acid and eicosapentaenoic acid ranged from 3.5 to 4.5% and 0.7% to 0.8%, respectively. The fatty acid profile and total fatty acids varied significantly under different nutrient deficiency. The greatest variation was found for oleic acid (9–46%) and linolenic acid (2–13%). The percentage of arachidonic acid was the highest with a standard nitrates concentration in the medium (5%) and the lowest in the absence of phosphates and nitrates (1.3–1.5%), but the absolute content in dry biomass was similar in all variants of the experiment (6.5–9.3 mg g −1 dry weight). The absence of nitrogen and both nitrogen and phosphorus led to a 3–fold increase in TFA in comparison with the control. Thus, this strain can be considered in biotechnological application as a potential producer of the essential linoleic acid or oleic acid. [ABSTRACT FROM AUTHOR]

Microalgal biomass represents a potential third generation feedstock that could be utilised as a source of carbohydrates for fermentative production of a range of platform biochemicals. Identifying microalgal strains with high biomass and carbohydrate productivities while also being amenable to downstream processes is key in improving the feasibility of these processes. Utilising marine microalgae capable of growing in seawater will decrease reliance on freshwater resources and improve the sustainability of production. This study screened several marine microalgae believed to accumulate carbohydrates to find new high performing strains. Four strains had high growth rates and accumulated carbohydrates >35% DW under stress. The strain Chlorella salina demonstrated the highest biomass and carbohydrate productivity, and alkaline autoflocculation (4 mM NaOH) enabled biomass recoveries >95% efficiency, resulting in an 8–10 × concentration of the culture. Under nutrient replete conditions, biomass productivity reached 0.6 g L −1  d −1 , significantly greater than that of nitrogen starved cultures. However, nitrogen starvation rapidly increased carbohydrate content to >50% DW in 2 days, resulting in carbohydrate productivities >0.20 g L −1  d −1 . Chlorella salina partitions the products of photosynthesis preferentially into carbohydrate synthesis under nitrogen starvation. A greater understanding of cellular physiology and carbon partitioning in response to nutrient stress will enable better control and optimisation of the bio-processes. This study has identified a potentially high performance marine microalga for carbohydrate production that is also amenable to low-cost harvesting. [ABSTRACT FROM AUTHOR]

Diatoms are a hugely diverse microalgal class, which possesses unique biological features and complex metabolic pathways and may activate sophisticated mechanisms to respond to environmental changes. Abiotic stress factors may limit growth rate of diatoms, but may also trigger intracellular signaling pathways that cause cells to undergo programmed cell death (PCD). Here we investigate the gene expression of different target genes related to cell death, namely programmed cell death 4 (PDCD4), tumor susceptibility gene 101 (TSG101), developmental and cell death (DCD) domain, death specific protein (DSP) and metacaspase (MC), using RT-qPCR in the cosmopolitan coastal centric diatom species Skeletonema marinoi , which contributes significantly to phytoplankton blooms in temperate waters. To this end, we undertook a detailed study of the best reference genes to analyze gene expression in S. marinoi under different experimental conditions (i.e. in different growth phases or under silica starvation). Results showed that DSP gene expression had a clear and constant increase along the S. marinoi growth curve reaching its maximum during the senescent phase. On the contrary, PDCD4, DCD, TSG101 and MC did not show any significant variation. These findings indicate that the DSP gene is a possible PCD marker induced by aging in this diatom species. In contrast, levels of DSP transcripts induced by silica starvation were relatively low compared to those induced by cell aging suggesting differential activation and/or regulation of the PCD machinery in response to different stressful conditions. Our study also expands the list of reference genes available for the diatom S. marinoi for normalization of RT-qPCR data of cells cultivated under different growth phases or under silica starvation. [ABSTRACT FROM AUTHOR]

Abstract: In microalgal cultures, most analyses of cellular processes are done in bulk, on the entire population of cells. Information gained from this is representative of the mean; however, it obscures the richness of cell-to-cell variation, which is a well-documented phenomenon. Using imaging flow cytometry, we evaluate changes in triacylglycerol (TAG) content and chlorophyll resulting from silicon or nitrogen deprivation in the diatom Cyclotella cryptica. This approach allows detailed interrogation of large numbers of individual cells and reveals cell-to-cell variation. This study demonstrates several previously undescribed phenomena related to TAG accumulation in microalgae. First, the rate of TAG accumulation varies over time, with a faster rate occurring at the latter stage of the process, resulting in hyperaccumulation in which the majority of the cell volume is comprised of lipid droplets. In C. cryptica and other diatoms this hyperaccumulation occurs strictly under autotrophic conditions. Second, there are distinct responses to silicon or nitrogen limitation, and variation within a given type of limitation treatment. Under most conditions there is a large spread in the population when measuring either chlorophyll or TAG. Heterogeneity within the total population indicates that caution should be taken in interpreting bulk measurements for a variety of variables (TAG, transcript, protein, metabolites, etc.) related to cellular responses. However, a potential means to couple subpopulation-level responses with bulk analysis approaches is described, which could take advantage of the nuances observed during the TAG accumulation process. [Copyright &y& Elsevier]

Abstract: Rice blast is one of the world’s most serious plant diseases and a significant threat to global food security. To invade rice plants, the blast fungus Magnaporthe oryzae forms specialised cells called appressoria that generate pressure and physical force to rupture the leaf cuticle. Recent evidence suggests that appressorium development is controlled by cell cycle checkpoints and involves autophagy. This culminates in programmed cell death of the fungal conidium and recycling of its contents to the appressorium. M. oryzae has evolved specialised regulatory mechanisms to respond to the nutrient-free environment of the rice leaf surface and its physical and chemical characteristics, in order to gain entry to the plant [Copyright &y& Elsevier]