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Establishment of the forward genetic analysis of the chlorophyll d-dominated cyanobacterium Acaryochloris marina MBIC 11017 by applying in vivo transposon mutagenesis system

Photosynthesis Research volume 125, pages 255–265 (2015)Cite this article

Abstract

Acaryochloris marina MBIC 11017 possesses chlorophyll (Chl) d as a major Chl, which enables this organism to utilize far-red light for photosynthesis. Thus, the adaptation mechanism of far-red light utilization, including Chl d biosynthesis, has received much attention, though a limited number of reports on this subject have been published. To identify genes responsible for Chl d biosynthesis and adaptation to far-red light, molecular genetic analysis of A. marina was required. We developed a transformation system for A. marina and introduced expression vectors into A. marina. In this study, the high-frequency in vivo transposon mutagenesis system recently established by us was applied to A. marina. As a result, we obtained mutants with the transposon in their genomic DNA at various positions. By screening transposon-tagged mutants, we isolated a mutant (Y1 mutant) that formed a yellow colony on agar medium. In the Y1 mutant, the transposon was inserted into the gene encoding molybdenum cofactor biosynthesis protein A (MoaA). The Y1 mutant was functionally complemented by introducing the moaA gene or increasing the ammonium ion in the medium. These results indicate that the mutation of the moaA gene reduced nitrate reductase activity, which requires molybdenum cofactor, in the Y1 mutant. This is the first successful forward genetic analysis of A. marina, which will lead to the identification of genes responsible for adaptation to far-red light.

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Abbreviations

aadA :

Streptomycin/spectinomycin resistance gene

Chl:

Chlorophyll

PCR:

Polymerase chain reaction

Em:

Erythromycin

HPLC:

High performance liquid chromatography

PS:

Photosystem

Sm:

Streptomycin

Sp:

Spectinomycin

Tnp:

Transposase

WT:

Wild-type strain

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Acknowledgments

This work was supported by JSPS KAKENHI Grant Numbers 22370017, 24658080, 26440141.

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Affiliations

  1. Graduate School of Human and Environmental Studies, Kyoto University, Kyoto, 606-8501, Japan

    Kazuyuki Watabe, Mamoru Mimuro & Tohru Tsuchiya

  2. Graduate School of Global Environmental Studies, Kyoto University, Kyoto, 606-8501, Japan

    Tohru Tsuchiya

Authors
  1. Kazuyuki Watabe
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  2. Mamoru Mimuro
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  3. Tohru Tsuchiya
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Corresponding author

Correspondence to Tohru Tsuchiya.

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Watabe, K., Mimuro, M. & Tsuchiya, T. Establishment of the forward genetic analysis of the chlorophyll d-dominated cyanobacterium Acaryochloris marina MBIC 11017 by applying in vivo transposon mutagenesis system. Photosynth Res 125, 255–265 (2015). https://doi.org/10.1007/s11120-015-0082-4

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  • DOI: https://doi.org/10.1007/s11120-015-0082-4

Keywords

  • Acaryochloris marina MBIC 11017
  • Chlorophyll d
  • Cyanobacteria
  • Genetic functional complementation
  • Transposon mutagenesis system