Skip to main content
NCBI home page
The Plant Cell logoLink to The Plant Cell
. 1993 Dec;5(12):1853–1863. doi: 10.1105/tpc.5.12.1853

Synechocystis sp PCC 6803 strains lacking photosystem I and phycobilisome function.

G Shen 1, S Boussiba 1, W F Vermaas 1
PMCID: PMC160410  PMID: 8305875

Abstract

To design an in vivo system allowing detailed analysis of photosystem II (PSII) complexes without significant interference from other pigment complexes, part of the psaAB operon coding for the core proteins of photosystem I (PSI) and part of the apcE gene coding for the anchor protein linking the phycobilisome to the thylakoid membrane were deleted from the genome of the cyanobacterium Synechocystis sp strain PCC 6803. Upon transformation and segregation at low light intensity (5 microE m-2 sec-1), a PSI deletion strain was obtained that is light tolerant and grows reasonably well under photoheterotrophic conditions at 5 microE m-2 sec-1 (doubling time approximately 28 hr). Subsequent inactivation of apcE by an erythromycin resistance marker led to reduction of the phycobilin-to-chlorophyll ratio and to a further decrease in light sensitivity. The resulting PSI-less/apcE- strain grew photoheterotrophically at normal light intensity (50 microE m-2 sec-1) with a doubling time of 18 hr. Deletion of apcE in the wild type resulted in slow photoautotrophic growth. The remaining phycobilins in apcE- strains were inactive in transferring light energy to PSII. Cells of both the PSI-less and PSI-less/apcE- strains had an approximately sixfold enrichment of PSII on a chlorophyll basis and were as active in oxygen evolution (on a per PSII basis) as the wild type at saturating light intensity. Both PSI-less strains described here are highly appropriate both for detailed PSII studies and as background strains to analyze site- and region-directed PSII mutants in vivo.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson S. L., McIntosh L. Light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. strain PCC 6803: a blue-light-requiring process. J Bacteriol. 1991 May;173(9):2761–2767. doi: 10.1128/jb.173.9.2761-2767.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Capuano V., Braux A. S., Tandeau de Marsac N., Houmard J. The "anchor polypeptide" of cyanobacterial phycobilisomes. Molecular characterization of the Synechococcus sp. PCC 6301 apce gene. J Biol Chem. 1991 Apr 15;266(11):7239–7247. [PubMed] [Google Scholar]
  3. Elhai J., Wolk C. P. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers. Gene. 1988 Aug 15;68(1):119–138. doi: 10.1016/0378-1119(88)90605-1. [DOI] [PubMed] [Google Scholar]
  4. Houmard J., Capuano V., Colombano M. V., Coursin T., Tandeau de Marsac N. Molecular characterization of the terminal energy acceptor of cyanobacterial phycobilisomes. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2152–2156. doi: 10.1073/pnas.87.6.2152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kim S., Sandusky P., Bowlby N. R., Aebersold R., Green B. R., Vlahakis S., Yocum C. F., Pichersky E. Characterization of a spinach psbS cDNA encoding the 22 kDa protein of photosystem II. FEBS Lett. 1992 Dec 7;314(1):67–71. doi: 10.1016/0014-5793(92)81463-v. [DOI] [PubMed] [Google Scholar]
  6. Kirilovsky D. L., Boussac A. G., van Mieghem F. J., Ducruet J. M., Sétif P. R., Yu J. J., Vermaas W. F., Rutherford A. W. Oxygen-evolving photosystem II preparation from wild type and photosystem II mutants of Synechocystis sp. PCC 6803. Biochemistry. 1992 Feb 25;31(7):2099–2107. doi: 10.1021/bi00122a030. [DOI] [PubMed] [Google Scholar]
  7. Mannan R. M., Whitmarsh J., Nyman P., Pakrasi H. B. Directed mutagenesis of an iron-sulfur protein of the photosystem I complex in the filamentous cyanobacterium Anabaena variabilis ATCC 29413. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10168–10172. doi: 10.1073/pnas.88.22.10168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Montoya G., Yruela I., Picorel R. Pigment stoichiometry of a newly isolated D1-D2-Cyt b559 complex from the higher plant Beta vulgaris L. FEBS Lett. 1991 Jun 3;283(2):255–258. doi: 10.1016/0014-5793(91)80601-x. [DOI] [PubMed] [Google Scholar]
  9. Nilsson F., Andersson B., Jansson C. Photosystem II characteristics of a constructed Synechocystis 6803 mutant lacking synthesis of the D1 polypeptide. Plant Mol Biol. 1990 Jun;14(6):1051–1054. doi: 10.1007/BF00019402. [DOI] [PubMed] [Google Scholar]
  10. Riethman H. C., Sherman L. A. Immunological Characterization of Iron-Regulated Membrane Proteins in the Cyanobacterium Anacystis nidulans R2. Plant Physiol. 1988 Oct;88(2):497–505. doi: 10.1104/pp.88.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Riethman H. C., Sherman L. A. Purification and characterization of an iron stress-induced chlorophyll-protein from the cyanobacterium Anacystis nidulans R2. Biochim Biophys Acta. 1988 Sep 14;935(2):141–151. doi: 10.1016/0005-2728(88)90211-3. [DOI] [PubMed] [Google Scholar]
  12. Smart L. B., McIntosh L. Genetic inactivation of the psaB gene in Synechocystis sp. PCC 6803 disrupts assembly of photosystem I. Plant Mol Biol. 1993 Jan;21(1):177–180. doi: 10.1007/BF00039628. [DOI] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES