scholarly journals Molecular Analysis of theIn SituGrowth Rates of Subsurface Geobacter Species

2012 ◽  
Vol 79 (5) ◽  
pp. 1646-1653 ◽  
Author(s):  
Dawn E. Holmes ◽  
Ludovic Giloteaux ◽  
Melissa Barlett ◽  
Milind A. Chavan ◽  
Jessica A. Smith ◽  
...  

ABSTRACTMolecular tools that can provide an estimate of thein situgrowth rate ofGeobacterspecies could improve understanding of dissimilatory metal reduction in a diversity of environments. Whole-genome microarray analyses of a subsurface isolate ofGeobacter uraniireducens, grown under a variety of conditions, identified a number of genes that are differentially expressed at different specific growth rates. Expression of two genes encoding ribosomal proteins,rpsCandrplL, was further evaluated with quantitative reverse transcription-PCR (qRT-PCR) in cells with doubling times ranging from 6.56 h to 89.28 h. Transcript abundance ofrpsCcorrelated best (r2= 0.90) with specific growth rates. Therefore, expression patterns ofrpsCwere used to estimate specific growth rates ofGeobacterspecies during anin situuranium bioremediation field experiment in which acetate was added to the groundwater to promote dissimilatory metal reduction. Initially, increased availability of acetate in the groundwater resulted in higher expression ofGeobacter rpsC, and the increase in the number ofGeobactercells estimated with fluorescentin situhybridization compared well with specific growth rates estimated from levels ofin situ rpsCexpression. However, in later phases, cell number increases were substantially lower than predicted fromrpsCtranscript abundance. This change coincided with a bloom of protozoa and increased attachment ofGeobacterspecies to solid phases. These results suggest that monitoringrpsCexpression may better reflect the actual rate thatGeobacterspecies are metabolizing and growing duringin situuranium bioremediation than changes in cell abundance.

2012 ◽  
Vol 78 (19) ◽  
pp. 7132-7136 ◽  
Author(s):  
Christian Dusny ◽  
Frederik Sven Ole Fritzsch ◽  
Oliver Frick ◽  
Andreas Schmid

ABSTRACTSingularized cells ofPichia pastoris,Hansenula polymorpha, andCorynebacterium glutamicumdisplayed specific growth rates under chemically and physically constant conditions that were consistently higher than those obtained in populations. This highlights the importance of single-cell analyses by uncoupling physiology and the extracellular environment, which is now possible using the Envirostat 2.0 concept.


2005 ◽  
Vol 50 (2) ◽  
pp. 230-241 ◽  
Author(s):  
Josefina García-Cantizano ◽  
Emilio O. Casamayor ◽  
Josep M. Gasol ◽  
Ricardo Guerrero ◽  
Carlos Pedrós-Alió

2021 ◽  
Vol 8 ◽  
Author(s):  
Najwa Al-Otaibi ◽  
Francisca C. García ◽  
Xosé Anxelu G. Morán

The diel variability of the abundance and cell size of picoplanktonic groups in the central Red Sea was monitored every 2 h in situ on 4 occasions (once per season) from 2015 to 2016. We distinguished Prochlorococcus, low (LF-Syn) and high (HF-Syn) fluorescence Synechococcus, small (Speuk) and large (Lpeuk) picoeukaryotes and two groups of heterotrophic prokaryotes of low (LNA) and high (HNA) nucleic acid content. The diel variability in abundance was less marked than in cell size and more apparent in autotrophs than heterotrophs. Specific growth rates were estimated by an empirical relationship from measurements obtained in bottle incubations of surface and deep samples collected in the winter compared with in situ variations in cell size over 24 h. Autotrophic picoplankton groups generally grew faster (0.23–0.77 d–1) than heterotrophic prokaryotes (0.12–0.50 d–1). Surface to 100 m depth-weighted specific growth rates displayed a clear seasonal pattern for Prochlorococcus, with maxima in winter (0.77 ± 0.07 d–1) and minima in fall (0.52 ± 0.07 d–1). The two groups of Synechococcus peaked in spring, with slightly higher growth rates of LF-Syn (0.57 ± 0.04 d–1) than HF-Syn (0.43 ± 0.04 d–1). Speuk and Lpeuk showed different seasonal patterns, with lower values of the former (0.27 ± 0.02 and 0.37 ± 0.04 d–1, respectively). HNA consistently outgrew LNA heterotrophic prokaryotes, with a higher growth in the epipelagic (0–200 m, 0.36 ± 0.03 d–1) than in the mesopelagic (200–700 m, 0.26 ± 0.03 d–1), while no differences were found for LNA cells (0.19 ± 0.03 d–1 and 0.17 ± 0.02 d–1, respectively). With all data pooled, the mean diel abundances of autotrophic picoplankton in the upper epipelagic and of HNA cells in the epipelagic and mesopelagic layers were significantly correlated with the specific growth rates estimated from cell size variations. Our high-resolution sampling dataset suggests that changes in growth rates underlie the noticeable seasonality of picoplankton recently described in these tropical waters.


2016 ◽  
Vol 82 (15) ◽  
pp. 4570-4583 ◽  
Author(s):  
Corinna Rebnegger ◽  
Tim Vos ◽  
Alexandra B. Graf ◽  
Minoska Valli ◽  
Jack T. Pronk ◽  
...  

ABSTRACTThe yeastPichia pastorisis a widely used host for recombinant protein production. Understanding its physiology at extremely low growth rates is a first step in the direction of decoupling product formation from cellular growth and therefore of biotechnological relevance. Retentostat cultivation is an excellent tool for studying microbes at extremely low specific growth rates but has so far not been implemented forP. pastoris. Retentostat feeding regimes were based on the maintenance energy requirement (mS) and maximum biomass yield on glucose (YX/Smax) estimated from steady-state glucose-limited chemostat cultures. Aerobic retentostat cultivation enabled reproducible, smooth transitions from a specific growth rate (μ) of 0.025 h−1to near-zero specific growth rates (μ < 0.001 h−1). At these near-zero specific growth rates, viability remained at least 97%. The value ofmSat near-zero growth rates was 3.1 ± 0.1 mg glucose per g biomass and h, which was 3-fold lower than themSestimated from faster-growing chemostat cultures. This difference indicated thatP. pastorisreduces its maintenance energy requirement at extremely low μ, a phenomenon not previously observed in eukaryotes. Intracellular levels of glycogen and trehalose increased, while μ progressively declined during retentostat cultivation. Transcriptional reprogramming toward zero growth included the upregulation of many transcription factors as well as stress-related genes and the downregulation of cell cycle genes. This study underlines the relevance of comparative analysis of maintenance energy metabolism, which has an important impact on large-scale industrial processes.IMPORTANCEThe yeastPichia pastorisnaturally lives on trees and can utilize different carbon sources, among them glucose, glycerol, and methanol. In biotechnology, it is widely used for the production of recombinant proteins. For both the understanding of life in its natural habitat and optimized production processes, a better understanding of cell physiology at an extremely low growth rate would be of extraordinary value. Therefore, we have grownP. pastorisin a retentostat, which allows the cultivation of metabolically active cells even at zero growth. Here we reached doubling times as long as 38 days and found thatP. pastorisdecreases its maintenance energy demand 3-fold during very slow growth, which enables it to survive with a much lower substrate supply than baker's yeast.


2013 ◽  
Vol 79 (8) ◽  
pp. 2503-2511 ◽  
Author(s):  
Rudolf Schneebeli ◽  
Thomas Egli

ABSTRACTMembers of the genusListeriaare fastidious bacteria with respect to their nutritional requirements, and several minimal media described in the literature fail to support growth of allListeriaspp. Furthermore, strict limitation by a single nutrient, e.g., the carbon source, has not been demonstrated for any of the published minimal media. This is an important prerequisite for defined studies of growth and physiology, including “omics.” Based on a theoretical analysis of previously published mineral media forListeria, an improved, well-balanced growth medium was designed. It supports the growth, not only of all testedListeria monocytogenesstrains, but of all otherListeriaspecies, with the exception ofL. ivanovii. The growth performance ofL. monocytogenesstrain Scott A was tested in the newly designed medium; glucose served as the only carbon and energy source for growth, whereas neither the supplied amino acids nor the buffering and complexing components (MOPS [morpholinepropanesulfonic acid] and EDTA) supported growth. Omission of amino acids, trace elements, or vitamins, alone or in combination, resulted in considerably reduced biomass yields. Furthermore, we monitored the specific growth rates of variousListeriastrains cultivated in the designed mineral medium and compared them to growth in complex medium (brain heart infusion broth [BHI]). The novel mineral medium was optimized for the commonly used strainL. monocytogenesScott A to achieve optimum cell yields and maximum specific growth rates. This mineral medium is the first published synthetic medium forListeriathat has been shown to be strictly carbon (glucose) limited.


Author(s):  
Benni Winding Hansen ◽  
Frank Jensen

Protozooplankton growth was measured by incubation in a walk-in cooling room at 5°C of filtered seawater from six stations on a south–north transect from open water through drift ice and into fast ice in the central Barents Sea (72°30′N–76°32′N). Eight species of naked ciliates, two species of tintinnids, seven species of athecate dinoflagellates and nine species of thecate dinoflagellates grew in the 24 and 48 h bottle incubations. Maximum potential mean specific growth rates of 0.84, 0.59, 0.39 and 0.39 d−1, respectively, indicated a hierarchy in growth rates determined by taxonomic differences where growth of naked ciliates > tintinnids > athecate dinoflagellates > thecate dinoflagellates. The mean±SD in situ temperature (4 to −1.8°C) corrected growth rates (Q10=2.8) for the six stations (0.74±0.31, 0.74±0.16, 0.25±0.16, 0.23±0.27 d−1) also suggested a higher growth for ciliates in comparison to dinoflagellates. Additionally, it revealed that the naked ciliates responded to the increased food availability, whereas this was not the case for all other groups of protozooplankton.


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