Chlorophyll <i>a</i> and NADPH Fluorescence Lifetimes in the Microalgae <i>Haematococcus pluvialis</i> (Chlorophyceae) under Normal and Astaxanthin-Accumulating Conditions

Kristoffersen, Arne S.; Svensen, Øyvind; Ssebiyonga, Nicolausi; Erga, Svein Rune; Stamnes, Jakob J.; Frette, Øyvind

doi:10.1366/12-06634

Cited by 11 publications

(12 citation statements)

References 55 publications

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“…Chemically closing the reaction centres of the green microalgae H. pluvialis has been shown to result in an increase in lifetime from 250 ps under normal circumstances to 420 ps (Kristoffersen et al. 2012 ) for DCMU-inhibited cells, and our present measurements on DCMU-inhibited Tetraselmis cells showed a lifetime of 425 ps. Our results for UV-stressed conditions reveal a slightly shorter lifetime at 390 ps.…”

Section: Resultssupporting

confidence: 67%

“…Closing the reaction centres by infiltration of DCMU (and thus shutting down photochemistry) has been shown to increase the fluorescence lifetime of the green algae H. pluvialis to around 420 ps (Kristoffersen et al. 2012 ), which is in accordance with the before-mentioned consequence: removing a de-excitation pathway results in an increased fluorescence lifetime. A second (slower) lifetime component often varies between 0.7 and 2 ns and is usually attributed to charge stabilisation and recombination in PSII reaction centres as well as closed or damaged reaction centres (Gilmore et al.…”

Section: Introductionsupporting

confidence: 65%

“… 2012 ; Kristoffersen et al. 2012 ). An ultra-short component at about 65 ps is attributed to PSI reaction centres.…”

Section: Introductionmentioning

confidence: 99%

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Chlorophyll a fluorescence lifetime reveals reversible UV-induced photosynthetic activity in the green algae Tetraselmis

et al. 2015

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The fluorescence lifetime is a very useful parameter for investigating biological materials on the molecular level as it is mostly independent of the fluorophore concentration. The green alga Tetraselmis blooms in summer, and therefore its response to UV irradiation is of particular interest. In vivo fluorescence lifetimes of chlorophyll a were measured under both normal and UV-stressed conditions of Tetraselmis. Fluorescence was induced by two-photon excitation using a femtosecond laser and laser scanning microscope. The lifetimes were measured in the time domain by time-correlated single-photon counting. Under normal conditions, the fluorescence lifetime was 262 ps, while after 2 h of exposure to UV radiation the lifetime increased to 389 ps, indicating decreased photochemical quenching, likely caused by a damaged and down-regulated photosynthetic apparatus. This was supported by a similar increase in the lifetime to 425 ps when inhibiting photosynthesis chemically using DCMU. Furthermore, the UV-stressed sample was dark-adapted overnight, resulting in a return of the lifetime to 280 ps, revealing that the damage caused by UV radiation is repairable on a relatively short time scale. This reversal of photosynthetic activity was also confirmed by measurements.

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Section: Resultssupporting

confidence: 67%

Section: Introductionsupporting

confidence: 65%

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Chlorophyll a fluorescence lifetime reveals reversible UV-induced photosynthetic activity in the green algae Tetraselmis

et al. 2015

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“…In biological samples, two (or more) lifetime components are often present, and therefore it is important to verify that lifetimes of a mixed substance will produce double-exponential decays resulting in the same lifetimes as the separate fluorophores yield from single-exponential decay. We have recently performed in vivo fluorescence lifetime measurements on microalgae with the same setup described in this paper[16], and we experienced that information on lifetime standards performed for two-photon experiments would be beneficial.…”

Section: Introductionmentioning

confidence: 99%

Testing Fluorescence Lifetime Standards using Two-Photon Excitation and Time-Domain Instrumentation: Rhodamine B, Coumarin 6 and Lucifer Yellow

et al. 2014

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Having good information about fluorescence lifetime standards is essential for anyone performing lifetime experiments. Using lifetime standards in fluorescence spectroscopy is often regarded as a straightforward process, however, many earlier reports are limited in terms of lifetime concentration dependency, solvents and other technical aspects. We have investigated the suitability of the fluorescent dyes rhodamine B, coumarin 6, and lucifer yellow as lifetime standards, especially to be used with two-photon excitation measurements in the time-domain. We measured absorption and emission spectra for the fluorophores to determine which wavelengths we should use for the excitation and an appropriate detector range. We also measured lifetimes for different concentrations, ranging from 10−2– 10−6 M, in both water, ethanol and methanol solutions. We observed that rhodamine B lifetimes depend strongly on concentration. Coumarin 6 provided the most stable lifetimes, with a negligible dependency on concentration and solvent. Lucifer yellow lifetimes were also found to depend little with concentration. Finally, we found that a mix of two fluorophores (rhodamine B/coumarin 6, rhodamine B/lucifer yellow, and coumarin 6/lucifer yellow) all yielded very similar lifetimes from a double-exponential decay as the separate lifetimes measured from a single-exponential decay. All lifetime measurements were made using two-photon excitation and obtaining lifetime data in the time-domain using time-correlated single-photon counting.

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“…Considering that the cell proliferation and biomass accumulation of OR are reduced [4], the purpose of maintaining such a high photosynthetic activity is unknown. Recent study by Kristoffersen et al by measuring in vivo fluorescence lifetimes of chlorophyll- a and NADPH demonstrated that open chlorophyll reaction centers did not change significantly in astaxanthin-accumulating cells; NADPH was necessary for astaxanthin synthesis in red-stage cells and might involve in photoprotection in stressed cells [5]. Additionally, the impairment in thylakoid membrane structure of nonmotile cells [6] and significant thylakoid membranes alterations in red cells [7] were also previously observed.…”

Section: Introductionmentioning

confidence: 84%

Comparison of Different Cells of Haematococcus pluvialis Reveals an Extensive Acclimation Mechanism during its Aging Process: From a Perspective of Photosynthesis

Xie

Gao

et al. 2013

PLoS ONE

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Both biomass dominated green vegetative cells (GV) and astaxanthin-dominated orange resting cells (OR) affect the final astaxanthin yield in industry. Examination of Haematococcus pluvialis revealed that the OR cells greatly varied from the GV cells at both cellular and subcellular levels. In particular, the thylakoid membranes in the OR were disassembled and fragmented. Furthermore, the OR conserved most of the photosynthetic pigments, with elevated concentrations of violaxanthin, antheraxanthin, and neoxanthin. Notably, moderate photosynthesis was detected in OR, even though most of the thylakoid membranes were disassembled, when compared with those in the GV. However, the energy distribution pattern between photosystem I and II (PSI and PSII) in the OR favored PSI, which was also confirmed by 77-K fluorescence. As zeaxanthin was not detected in the OR, we attribute the acclimation role to astaxanthin, instead of xanthophyll cycle. Additionally, proteomic-scale comparison analysis of thylakoids of the OR and GV indicated no photosynthetically remarkable variations. However, an extensive acclimation mechanism of H. pluvialis was proposed, in which proteins in thylakoid of GV were noted to be involved in biomass accumulation and those in OR were involved in stress response. Conclusions of the comparative analysis might provide some physiological background of OR for astaxanthin production by using H. pluvialis.

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Chlorophyll a and NADPH Fluorescence Lifetimes in the Microalgae Haematococcus pluvialis (Chlorophyceae) under Normal and Astaxanthin-Accumulating Conditions

Cited by 11 publications

References 55 publications

Chlorophyll a fluorescence lifetime reveals reversible UV-induced photosynthetic activity in the green algae Tetraselmis

Chlorophyll a fluorescence lifetime reveals reversible UV-induced photosynthetic activity in the green algae Tetraselmis

Testing Fluorescence Lifetime Standards using Two-Photon Excitation and Time-Domain Instrumentation: Rhodamine B, Coumarin 6 and Lucifer Yellow

Comparison of Different Cells of Haematococcus pluvialis Reveals an Extensive Acclimation Mechanism during its Aging Process: From a Perspective of Photosynthesis

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