Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria

Zavřel, Tomáš; Sinetova, Maria A.; Červený, Jan

doi:10.21769/bioprotoc.1467

Cited by 121 publications

(70 citation statements)

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“…Content of chlorophyll a, carotenoids and phycobilisomes was measured spectrophotometrically following the protocols of Zavřel et al (2015a) and Zavřel et al (2018a).…”

Section: Pigment Content Measurementsmentioning

confidence: 99%

Quantitative insights into the cyanobacterial cell economy

Zavřel

Faizi

Loureiro

et al. 2018

Preprint

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Phototrophic microorganisms are promising resources for green biotechnology. 13 Compared to heterotrophic microorganisms, however, the cellular economy of phototrophic 14 growth is still insufficiently understood. We provide a quantitative analysis of light-limited, 15 light-saturated, and light-inhibited growth of the cyanobacterium Synechocystis sp. PCC 6803 using 16 a reproducible cultivation setup. We report key physiological parameters, including growth rate, cell 17 size, and photosynthetic activity over a wide range of light intensities. Intracellular proteins were 18 quantified to monitor proteome allocation as a function of growth rate. Among other physiological 19 adaptations, we identify an upregulation of the translational machinery and downregulation of light 20 harvesting components with increasing light intensity and growth rate. The resulting growth laws 21 are discussed in the context of a coarse-grained model of phototrophic growth and available data 22 obtained by a comprehensive literature search. Our insights into quantitative aspects of 23 cyanobacterial adaptations to different growth rates have implications to understand and optimize 24 photosynthetic productivity. 25 26 2014). While quantitative insight into the cellular economy of phototrophic microorganisms is 36 still scarce, the cellular economy of heterotrophic growth has been studied extensively-starting 37 with the seminal works of Monod, Neidhardt, and others (Neidhardt et al., 1990; Neidhardt, 1999; 38 Jun et al., 2018) to more recent quantitative studies of microbial resource allocation (Molenaar 39 et al.40 Maitra and Dill, 2015; Weiße et al., 2015). In response to changing environments, heterotrophic 41 1 of 28 Manuscript submitted to eLife microorganisms are known to differentially allocate their resources: with increasing growth rate, 42 heterotrophic microorganisms typically exhibit upregulation of ribosomes and other proteins 43 related to translation and protein synthesis (Scott et al., 2010; Molenaar et al., 2009; Peebo et al., 44 2015), exhibit complex changes in transcription profiles, e.g. (Klumpp et al., 2009; Matsumoto et al., 45 2013), and increase cell size (Kafri et al., 2016). The molecular limits of heterotrophic growth have 46 been described thoroughly (Kafri et al., 2016; Erickson et al., 2017; Scott et al., 2014; Metzl-Raz 47 et al., 2017; Klumpp et al., 2013). 48 In contrast, only few studies so far have addressed the limits of cyanobacterial growth from an 49 experimental perspective (Bernstein et al., 2016; Yu et al., 2015; Abernathy et al., 2017; Ungerer 50 et al., 2018; Jahn et al., 2018). Of particular interest were the adaptations that enable fast pho-51 toautotrophic growth (Bernstein et al., 2016; Yu et al., 2015; Abernathy et al., 2017; Ungerer et al., 52 2018). The cyanobacterium with the highest known photoautotrophic growth rate, growing with a 53 doubling time of up to ∼ 1.5h, is the strain Synechococcus elongatus UTEX 2973 (Ungerer et al., 54 2018). Compared to its c...

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“…Content of chlorophyll a, carotenoids and phycobilisomes was measured spectrophotometrically following the protocols of Zavřel et al (2015a) and Zavřel et al (2018a).…”

Section: Pigment Content Measurementsmentioning

confidence: 99%

Quantitative insights into the cyanobacterial cell economy

Zavřel

Faizi

Loureiro

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

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“…Both pigment extractions were left overnight at 4°C before vortexing and centrifugation (15 min at 10,000 g; HERMLE Z233M-2, Germany). The absorbance of the methanol extract was read at 665 nm for chlorophyll a determination (Meeks & Castenholz, 1971) and at 470 and 720 nm for carotenoid determination (Wellburn, 1994;Zavřel, Sinetova, & Červený, 2015). The total phycobiliprotein content was determined from the absorption readings at 562, 615 and 652 nm (Bennett & Bogorad, 1971, 1973.…”

Section: Pigment Determinationsmentioning

confidence: 99%

An investigation into the effects of increasing salinity on photosynthesis in freshwater unicellular cyanobacteria during the late Archaean

Herrmann

Gehringer

2019

Geobiology

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The oldest species of bacteria capable of oxygenic photosynthesis today are the freshwater Cyanobacteria Gloeobacter spp., belonging to the class Oxyphotobacteria. Several modern molecular evolutionary studies support the freshwater origin of cyanobacteria during the Archaean and their subsequent acquisition of salt tolerance mechanisms necessary for their expansion into the marine environment. This study investigated the effect of a sudden washout event from a freshwater location into either a brackish or marine environment on the photosynthetic efficiency of two unicellular freshwater cyanobacteria: the salt‐tolerant Chroococcidiopsis thermalis PCC7203 and the cyanobacterial phylogenetic root species, Gloeobacter violaceus PCC7421. Strains were cultured under present atmospheric levels (PAL) of CO2 or an atmosphere containing elevated levels of CO2 and reduced O2 (eCO2rO2) in simulated shallow water or terrestrial environmental conditions. Both strains exhibited a reduction in growth rates and gross photosynthesis, accompanied by significant reductions in chlorophyll a content, in brackish water, with only C. thermalis able to grow at marine salinity levels. While the experimental atmosphere caused a significant increase in gross photosynthesis rates in both strains, it did not increase their growth rates, nor the amount of O2 released. The differences in growth responses to increasing salinities could be attributed to genetic differences, with C. thermalis carrying additional genes for trehalose synthesis. This study demonstrates that, if cyanobacteria did evolve in a freshwater environment, they would have been capable of withstanding a sudden washout into increasingly saline environments. Both C. thermalis and G. violaceus continued to grow and photosynthesise, albeit at diminished rates, in brackish water, thereby providing a route for the evolution of open ocean‐dwelling strains, necessary for the oxygenation of the Earth's atmosphere.

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“…In order to determine the exo-polysaccharide (EPS) content, an extract was taken from the fresh biomass using the formaldehyde-NaOH method as described by [26], and then the polysaccharide content in the extract was measured using the phenol-sulfuric acid method as described by [27]. To measure the photosynthetic pigments (chlorophyll a and carotenoids), extraction and spectrophotometric absorbance measurements were conducted using the protocol described by [28], and the chlorophyll a content was calculated from the absorbance value using the formula Chla [µg/mL] = 12.9447 (A665-A720) as described by [29]. Carotenoid content was calculated using the formula, carotenoids [µg/mL] = [1000 (A470-A720)-2.86 (Chla [µg/mL])]/221 as described by [30].…”

Section: Measurement Of Biomass Exo-polysaccharides (Epss) and Photomentioning

confidence: 99%

Exploring an Integrated Manure-Seawater System for Sustainable Cyanobacterial Biomass Production

Sido

2019

Applied Sciences

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Cyanobacterial biomass is important for biofuel and biofertilizer, however, biomass production requires expensive chemical growth nutrients. To address this issue, we explored the use of inexpensive growth nutrient media from an integrated manure-seawater system for cyanobacterial biomass production. Salt-tolerant cyanobacterial strain HSaC and salt-sensitive cyanobacterial strain LC were tested to evaluate the potential of integrated manure-seawater media for sustainable cyanobacterial biomass production. As a prerequisite for seawater experiments, strain HSaC was grown at different NaCl concentrations (0 mM, 60 mM, 120 mM, 180 mM, 240 mM and 300 mM) to identify the optimum salt concentration. The highest biomass yield and photosynthetic pigment contents were obtained at 120 mM NaCl concentration. The highest exo-polysaccharide (EPS) content was obtained at 180 mM NaCl concentration. The treatments for the manure-seawater media were cow manure, pig manure, chicken manure and BG11, each with distilled water, diluted seawater and non-diluted seawater. The highest biomass and photosynthetic pigment yield for cyanobacterial strains LC and HSaC were obtained from 0.5 dS/m and 10 dS/m diluted seawater integrated with cow manure, respectively, but pig and chicken manure performed poorly. Overall, the biomass production and photosynthetic pigment results from cow manure-seawater were relatively better than those from the reference media (BG11). Based on the current findings, it is concluded that the growth nutrients from integrated cow manure-seawater can wholly substitute for the BG11 without affecting cyanobacterial growth, thereby reducing the usage of expensive chemical growth media. Thus, the results of study help to enhance the biomass production of both salt-sensitive and salt-tolerant cyanobacteria for sustainable biofuel and biofertilizer production.

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Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria

Cited by 121 publications

References 4 publications

Quantitative insights into the cyanobacterial cell economy

Quantitative insights into the cyanobacterial cell economy

An investigation into the effects of increasing salinity on photosynthesis in freshwater unicellular cyanobacteria during the late Archaean

Exploring an Integrated Manure-Seawater System for Sustainable Cyanobacterial Biomass Production

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