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Planta

, Volume 230, Issue 1, pp 27–37 | Cite as

Analysis of the phytochrome gene family in Ceratodon purpureus by gene targeting reveals the primary phytochrome responsible for photo- and polarotropism

  • Franz Mittmann
  • Sven Dienstbach
  • Andrea Weisert
  • Christoph Forreiter
Original Article

Abstract

Using gene targeting by homologous recombination in Ceratodon purpureus, we were able to knock out four phytochrome photoreceptor genes independently and to analyze their function with respect to red light dependent phototropism, polarotropism, and chlorophyll content. The strongest phenotype was found in knock-out lines of a newly described phytochrome gene termed CpPHY4 lacking photo- and polarotropic responses at moderate fluence rates. Eliminating the atypical phytochrome gene CpPHY1, which is the only known phytochrome-like gene containing a putative C-terminal tyrosine kinase-like domain, affects red light-induced chlorophyll accumulation. This result was surprising, since no light dependent function was ever allocated to this unusual gene.

Keywords

Ceratodon Gene targeting Knock-out Phytochrome 

Abbreviations

CTAB

Cetyltrimethylammonium bromide

GT

Gene targeting

HR

Homologous recombination

ONP

Oligonucleotide primer

RAcE

Rapid amplification of cDNA ends

RT-PCR

Reverse transcription based polymerase chain reaction

Notes

Acknowledgments

We would like to thank Dennis Hauser, Frankfurt a. M., Germany, for providing his software “MossBendingPlotter 3.0” visualizing the bending angles presented in Figs. 4, 5, 6, Tanja Gans for technical assistance and Man Mohan (Stowers Institute for medical research, Kansas City, MO, USA) for critical reading the manuscript and helpful discussion. The work was funded by grants of the Deutsche Forschungsgemeinschaft to CF (FO-290/4-1).

Supplementary material

425_2009_922_MOESM1_ESM.pdf (32 kb)
Schematic plot of the four different phytochrome genes and the corresponding knock-out constructs. Exon sequences are highlighted in orange. Restriction sites relevant for Southern-blot analysis are indicated above the gene. Red bars indicate size and position of the labeled PCR probe used for Southern-blot analysis. Inserted hygromycin and G418 resistance cassettes are visualized green. a CpPHY1 and cpphy1 knock-out; b CpPHY2 and cpphy2 knock-out; c CpPHY3 and cpphy3 knock-out; d CpPHY4 and cpphy4 knock-out, respectively (PDF 31 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Franz Mittmann
    • 1
  • Sven Dienstbach
    • 1
    • 2
  • Andrea Weisert
    • 1
  • Christoph Forreiter
    • 1
  1. 1.Department of Plant PhysiologyJustus Liebig UniversityGiessenGermany
  2. 2.Department of GeneticsJustus Liebig UniversityGiessenGermany

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