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Inhibition of chlorophyll biosynthesis at the protochlorophyllide reduction step results in the parallel depletion of Photosystem I and Photosystem II in the cyanobacterium Synechocystis PCC 6803

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Abstract

In most oxygenic phototrophs, including cyanobacteria, two independent enzymes catalyze the reduction of protochlorophyllide to chlorophyllide, which is the penultimate step in chlorophyll (Chl) biosynthesis. One is light-dependent NADPH:protochlorophyllide oxidoreductase (LPOR) and the second type is dark-operative protochlorophyllide oxidoreductase (DPOR). To clarify the roles of both enzymes, we assessed synthesis and accumulation of Chl-binding proteins in mutants of cyanobacterium Synechocystis PCC 6803 that either completely lack LPOR or possess low levels of the active enzyme due to its ectopic regulatable expression. The LPOR-less mutant grew photoautotrophically in moderate light and contained a maximum of 20 % of the wild-type (WT) Chl level. Both Photosystem II (PSII) and Photosystem I (PSI) were reduced to the same degree. Accumulation of PSII was mostly limited by the synthesis of antennae CP43 and especially CP47 as indicated by the accumulation of reaction center assembly complexes. The phenotype of the LPOR-less mutant was comparable to the strain lacking DPOR that also contained <25 % of the wild-type level of PSII and PSI when cultivated under light-activated heterotrophic growth conditions. However, in the latter case, we detected no reaction center assembly complexes, indicating that synthesis was almost completely inhibited for all Chl-proteins, including the D1 and D2 proteins.

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Abbreviations

BN:

Blue native

Chl:

Chlorophyll

Chlide:

Chlorophyllide

CN:

Clear native

CP43a/CP43b:

Two forms of unassembled CP43

DPOR:

Dark-operative protochlorophyllide oxidoreductase

LAHG:

Light-activated heterotrophic growth

LL:

Low light

LPOR:

Light-dependent protochlorophyllide oxidoreductase

NL:

Normal light

Pchlide:

Protochlorophyllide

pD1/iD1:

Precursor/intermediate form of D1

PSI:

Photosystem I

PSI(1):

PSI monomer

PSI(3):

PSI trimer

PSII:

Photosystem II

RC:

PSII reaction center complex

RC* and RCa:

Reaction center complexes lacking antennae CP47 and CP43 and accumulating in mutants unable to synthesize CP47

RC47:

PSII monomer lacking CP43

RCC1:

PSII monomer

RCC2:

PSII dimer

RCCS1:

Supercomplex containing PSII and PSI proteins

U.P.:

Unassembled proteins

WT:

Wild-type

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Acknowledgments

We thank Henry Valentin for providing pCER20 plasmid, C. Neil Hunter (Sheffield University, UK) for anti-LPOR and Peter Nixon (Imperial College L ondon, UK) for the WT strain and the anti-YCF48 antibody. This work was supported by projects Algatech (CZ.1.05/2.1.00/03.0110), RVO61388971, by projects P501/10/1000 and P501/12/G055 of the Grant Agency of the Czech Republic, and by a project 073/2010/P-PřF of the Grant Agency of the University of South Bohemia.

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Authors and Affiliations

  1. Department of Phototrophic Microorganisms, Institute of Microbiology, Academy of Sciences, Opatovický mlýn, 37981, Třeboň, Czech Republic

    Jana Kopečná, Roman Sobotka & Josef Komenda

  2. Faculty of Science, University of South Bohemia, Branišovská 31, 370 05, České Budějovice, Czech Republic

    Jana Kopečná, Roman Sobotka & Josef Komenda

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  1. Jana Kopečná
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  2. Roman Sobotka
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  3. Josef Komenda
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Correspondence to Josef Komenda.

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A contribution to the Special Issue on Evolution and Biogenesis of Chloroplasts and Mitochondria.

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Kopečná, J., Sobotka, R. & Komenda, J. Inhibition of chlorophyll biosynthesis at the protochlorophyllide reduction step results in the parallel depletion of Photosystem I and Photosystem II in the cyanobacterium Synechocystis PCC 6803. Planta 237, 497–508 (2013). https://doi.org/10.1007/s00425-012-1761-4

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