Letter

Origin and function of stomata in the moss Physcomitrella patens

  • Nature Plants 2, Article number: 16179 (2016)
  • doi:10.1038/nplants.2016.179
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Abstract

Stomata are microscopic valves on plant surfaces that originated over 400 million years (Myr) ago and facilitated the greening of Earth's continents by permitting efficient shoot–atmosphere gas exchange and plant hydration1. However, the core genetic machinery regulating stomatal development in non-vascular land plants is poorly understood2,​3,​4 and their function has remained a matter of debate for a century5. Here, we show that genes encoding the two basic helix–loop–helix proteins PpSMF1 (SPEECH, MUTE and FAMA-like) and PpSCREAM1 (SCRM1) in the moss Physcomitrella patens are orthologous to transcriptional regulators of stomatal development in the flowering plant Arabidopsis thaliana and essential for stomata formation in moss. Targeted P. patens knockout mutants lacking either PpSMF1 or PpSCRM1 develop gametophytes indistinguishable from wild-type plants but mutant sporophytes lack stomata. Protein–protein interaction assays reveal heterodimerization between PpSMF1 and PpSCRM1, which, together with moss–angiosperm gene complementations6, suggests deep functional conservation of the heterodimeric SMF1 and SCRM1 unit is required to activate transcription for moss stomatal development, as in A. thaliana7. Moreover, stomata-less sporophytes of ΔPpSMF1 and ΔPpSCRM1 mutants exhibited delayed dehiscence, implying stomata might have promoted dehiscence in the first complex land-plant sporophytes.

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Acknowledgements

We thank R. Haas and T. Fulton for excellent technical assistance. R.S.C. was supported by a NERC studentship. D.C.B is a GBMF investigator of the Howard Hughes Medical Institute. D.J.B. acknowledges funding through an ERC Advanced Grant (CDREG, 322998). R.R. acknowledges funding through the Excellence Initiative of the German Federal and States Governments (EXC294). A.C.C. and Y.K. acknowledge support from BBSRC (Grant numbers BB/F001797/1 and BB/I006710/1).

Author information

Author notes

    • Caspar C. Chater
    •  & Robert S. Caine

    These authors contributed equally to this work.

Affiliations

  1. Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de Mexico, Cuernavaca 62210, Mexico

    • Caspar C. Chater
  2. Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK

    • Robert S. Caine
    • , Andrew Fleming
    •  & David J. Beerling
  3. Plant Biotechnology, Faculty of Biology, University of Freiburg, Schaenzlestraße 1, 79104 Freiburg, Germany

    • Marta Tomek
    • , Daniel Lang
    • , Eva L. Decker
    •  & Ralf Reski
  4. Royal College of Veterinary Surgeons, Belgravia House, 62–64 Horseferry Road, London SW1P 2AF, UK

    • Simon Wallace
  5. Centre for Plant Sciences, University of Leeds, Leeds LS2 9JT, UK

    • Yasuko Kamisugi
    •  & Andrew C. Cuming
  6. Department of Molecular Cellular and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109-1048, USA

    • Cora A. MacAlister
  7. Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK

    • Stuart Casson
    •  & Julie E. Gray
  8. HHMI and Department of Biology, Stanford University, Stanford, California 94305-5020, USA

    • Dominique C. Bergmann
  9. Plant Molecular Cell Biology, Faculty of Biology, Ludwig-Maximilians-Universität München, LMU Biocenter, Großhaderner Straße 2, 82152 Planegg-Martinsried, Germany

    • Wolfgang Frank
  10. BIOSS – Centre for Biological Signalling Studies, 79104 Freiburg, Germany

    • Ralf Reski

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Contributions

C.C.C., R.S.C., W.F., J.E.G., A.F., D.J.B. and R.R. designed the study, C.C.C., R.S.C., D.L. and M.T. undertook the experiments with contributions from S.W., Y.K. and A.C.C., C.A.M., S.C., D.C.B., D.L., E.L.D and W.F. contributed materials and advice. A.C.C. constructed vectors for moss targeted knockout (SMF1 and SMF2), Y.K. carried out moss transformation (SMF2-KO) and yeast two-hybrid analysis, A.C.C. and Y.K. carried out Southern blot hybridization of the knockout mutant lines. D.J.B., R.R, C.C.C. and R.S.C. wrote the paper with contributions from D.C.B. All authors read, commented on and approved the final version of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Ralf Reski or David J. Beerling.

Supplementary information

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    Supplementary Information

    Supplementary Figures 1–15, Supplementary Methods, Supplementary Discussion, Supplementary Tables 1 and 2, Supplementary References, Supplementary Data Files 1–4.

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    Supplementary Dataset 1.

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    Supplementary Data 2

    Supplementary Dataset 2.

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    Supplementary Data 4

    Supplementary Dataset 4.