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Regulation of photosynthesis during heterocyst differentiation in Anabaena sp. strain PCC 7120 investigated in vivo at single-cell level by chlorophyll fluorescence kinetic microscopy

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Abstract

Changes of photosynthetic activity in vivo of individual heterocysts and vegetative cells in the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120 during the course of diazotrophic acclimation were determined using fluorescence kinetic microscopy (FKM). Distinct phases of stress and acclimation following nitrogen step-down were observed. The first was a period of perception, in which the cells used their internally stored nitrogen without detectable loss of PS II activity or pigments. In the second, the stress phase of nitrogen limitation, the cell differentiation occurred and an abrupt decline of fluorescence yield was observed. This decline in fluorescence was not paralleled by a corresponding decline in photosynthetic pigment content and PS II activity. Both maximal quantum yield and sustained electron flow were not altered in vegetative cells, only in the forming heterocysts. The third, acclimation phase started first in the differentiating heterocysts with a recovery of PS II photochemical yields \(F_{\text{v}} /F_{\text{m}} ,\;F^{\prime}_{\text{v}} /F^{\prime}_{\text{m}}.\) Afterwards, the onset of nitrogenase activity was observed, followed by the restoration of antenna pigments in the vegetative cells, but not in the heterocysts. Surprisingly, mature heterocysts were found to have an intact PS II as judged by photochemical yields, but a strongly reduced PS II-associated antenna as judged by decreased F 0. The possible importance of the functional PS II in heterocysts is discussed. Also, the FKM approach allowed to follow in vivo and evaluate the heterogeneity in photosynthetic performance among individual vegetative cells as well as heterocysts in the course of diazotrophic acclimation. Some cells along the filament (so-called “superbright cells”) were observed to display transiently increased fluorescence yield, which apparently proceeded by apoptosis.

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Abbreviations

Chl:

Chlorophyll

FKM:

Fluorescence kinetic microscope as defined by Küpper et al. (2000)

F 0 :

Minimal fluorescence yield of a dark adapted sample in non-actinic measuring light

F m :

Maximum fluorescence yield of a dark adapted sample

F s :

Steady state fluorescence under actinic irradiance, after the end of the induction transient

F v :

F m − F 0 = Variable fluorescence

F p :

Fluorescence yield at the peak of the induction curve after the onset of actinic light

FY:

Fluorescence yield

NSD:

Nitrogen step-down

NPQ:

(F m − \(F^{\prime}_{\text{m}})\)/F m = Non-photochemical quenching

PBS:

Phycobilisomes

PS:

Photosystem

ΔF :

\(F^{\prime}_{\text{m}} -F^{\prime}_{\text{t}}\), i.e., the response of fluorescence yield to a saturating irradiation pulse in light acclimated state

ΦPSII :

\(F^{\prime}_{\text{m}} -F^{\prime}_{\text{t}}/F^{\prime}_{\text{m}}\) = Light-acclimated electron flow through PS II (Genty et al. 1989)

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Acknowledgments

The authors would like to thank Vit’a Lukes for help with data analysis. Financial support was provided by the GACR grant 206/08/1683 and by the project Algatech (CZ.1.05/2.1.00/03.0110). HK would like to thank the “Studienstiftung des Deutschen Volkes” for a fellowship during the initial part of this project.

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Authors and Affiliations

  1. Laboratory of Photosynthesis—Algatech, Institute of Microbiology, Academy of Sciences of the Czech Republic, Třeboň, 379 81, Czech Republic

    Naila Ferimazova, Kristina Felcmanová, Eva Šetlíková, Barbora Šedivá & Ondřej Prášil

  2. Faculty of Science, University of South Bohemia, Branišovská 31, Ceske Budejovice, 370 05, Czech Republic

    Naila Ferimazova, Kristina Felcmanová, Hendrik Küpper & Ondřej Prášil

  3. Fachbereich Biologie, Universität Konstanz, 78457, Konstanz, Germany

    Hendrik Küpper

  4. IMIT, Microbiology/Organismic Interactions, Department of Biology, University of Tübingen, Auf der Morgenstelle 28, 72076, Tübingen, Germany

    Iris Maldener

  5. Lehrstuhl für Zellbiologie und Pflanzenphysiologie, Universität Regensburg, 93040, Regensburg, Germany

    Günther Hauska

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  1. Naila Ferimazova
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  2. Kristina Felcmanová
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  3. Eva Šetlíková
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  4. Hendrik Küpper
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  8. Ondřej Prášil
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Correspondence to Ondřej Prášil.

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This paper is dedicated to Dr. Ivan Šetlík (1928–2009), our friend and mentor, who initiated this and several other studies on single-cell physiology of algae and diazotrophic cyanobacteria and was involved in the construction of the fluorescence kinetic microscope.

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Ferimazova, N., Felcmanová, K., Šetlíková, E. et al. Regulation of photosynthesis during heterocyst differentiation in Anabaena sp. strain PCC 7120 investigated in vivo at single-cell level by chlorophyll fluorescence kinetic microscopy. Photosynth Res 116, 79–91 (2013). https://doi.org/10.1007/s11120-013-9897-z

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