Advertisement

Observation of Phototropic Responses in the Liverwort Marchantia polymorpha

  • Aino Komatsu
  • Ryuichi Nishihama
  • Takayuki KohchiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1924)

Abstract

The liverwort species, Marchantia polymorpha, shows environment-dependent morphological plasticity throughout its life cycle. Thalli, representing the predominant body form throughout most of this bryophyte’s life cycle, grow with repeated dichotomous branching at the apex and develop horizontally under sufficient light intensity. Spores, after germination, produce a mass of cells, called sporelings, which then grow into thalli. Both thalli and sporelings, if grown under weak light conditions, form narrow shapes, and their apices grow toward the light source. These phototropic responses are specific to blue light and dependent on the blue-light receptor phototropin. This chapter provides several basic procedures, along with some tips, for designing and performing experiments with M. polymorpha to observe their phototropic responses, as well as methods for observing the localization of the phototropin “Mpphot” with a fluorescent protein tag.

Key words

Liverwort Marchantia polymorpha Phototropic responses Blue light Sporeling Gemmaling Phototropin 

Notes

Acknowledgments

We thank Prof. Masamitsu Wada (Tokyo Metropolitan University), Dr. Noriyuki Suetsugu (Kyoto University), and Dr. Akane Kubota (Kyoto University) for technical advice and helpful discussions.

References

  1. 1.
    Qiu YL, Li L, Wang B, Chen Z, Knoop V, Groth-Malonek M, Dombrovska O, Lee J, Kent L, Rest J, Estabrook GF, Hendry TA, Taylor DW, Testa CM, Ambros M, Crandall-Stotler B, Duff RJ, Stech M, Frey W, Quandt D, Davis CC (2006) The deepest divergences in land plants inferred from phylogenomic evidence. Proc Natl Acad Sci U S A 103:15511–15516CrossRefGoogle Scholar
  2. 2.
    Bowman JL, Floyd SK, Sakakibara K (2007) Green genes—comparative genomics of the green branch of life. Cell 129:229–234CrossRefGoogle Scholar
  3. 3.
    Bowman JL (2015) A brief history of Marchantia from Greece to genomics. Plant Cell Physiol 57:210–229CrossRefGoogle Scholar
  4. 4.
    Shimamura M (2016) Marchantia polymorpha: taxonomy, phylogeny and morphology of a model system. Plant Cell Physiol 57:230–256CrossRefGoogle Scholar
  5. 5.
    O'Hanlon ME (1926) Germination of spores and early stages in development of gametophyte of Marchantia polymorpha. Bot Gaz 82:215–222CrossRefGoogle Scholar
  6. 6.
    Nakazato T, Kadota A, Wada M (1999) Photoinduction of spore germination in Marchantia polymorpha L. is mediated by photosynthesis. Plant Cell Physiol 40:1014–1020CrossRefGoogle Scholar
  7. 7.
    Fredericq H, De Greef J (1968) Photomorphogenic and chlorophyll studies in the bryophyte Marchantia polymorpha. Physiol Plant 21:346–359CrossRefGoogle Scholar
  8. 8.
    De Greef J, Fredericq H (1969) Photomorphogenic and chlorophyll studies in the bryophyte Marchantia polymorpha. Physiol Plant 22:462–468CrossRefGoogle Scholar
  9. 9.
    Fredericq H, De Greef J (1966) Red (R), far-red (FR) photoreversible control of growth and chlorophyll content in light-grown thalli of Marchantia polymorpha L. Naturwissenschaften 53:337–337CrossRefGoogle Scholar
  10. 10.
    Rethy R, Fredericq H, De Greef J (1990) Light and the diagravitropic growth of Marchantia polymorpha L. thalli: phytochrome-controlled epinasty. Mem Soc R Bot Belg 12:77–88Google Scholar
  11. 11.
    Christie JM, Salomon M, Nozue K, Wada M, Briggs WR (1999) LOV (light, oxygen, or voltage) domains of the blue-light photoreceptor phototropin (nph1): binding sites for the chromophore flavin mononucleotide. Proc Natl Acad Sci U S A 96:8779–8783CrossRefGoogle Scholar
  12. 12.
    Christie JM, Suetsugu N, Sullivan S, Wada M (2018) Shining light on the function of NPH3/RPT2-like proteins in phototropin signaling. Plant Physiol 176:1015–1024CrossRefGoogle Scholar
  13. 13.
    Sakai T, Wada T, Ishiguro S, Okada K (2000) RPT2. A signal transducer of the phototropic response in Arabidopsis. Plant Cell 12:225–236CrossRefGoogle Scholar
  14. 14.
    Motchoulski A, Liscum E (1999) Arabidopsis NPH3: a NPH1 photoreceptor-interacting protein essential for phototropism. Science 286:961–964CrossRefGoogle Scholar
  15. 15.
    de Wit M, Galvão VC, Fankhauser C (2016) Light-mediated hormonal regulation of plant growth and development. Annu Rev Plant Biol 67:513–537CrossRefGoogle Scholar
  16. 16.
    Ishizaki K, Johzuka-Hisatomi Y, Ishida S, Iida S, Kohchi T (2013) Homologous recombination-mediated gene targeting in the liverwort Marchantia polymorpha L. Sci Rep 3:1532CrossRefGoogle Scholar
  17. 17.
    Chiyoda S, Ishizaki K, Kataoka H, Yamato KT, Kohchi T (2008) Direct transformation of the liverwort Marchantia polymorpha L. by particle bombardment using immature thalli developing from spores. Plant Cell Rep 27:1467–1473CrossRefGoogle Scholar
  18. 18.
    Kubota A, Ishizaki K, Hosaka M, Kohchi T (2013) Efficient Agrobacterium-mediated transformation of the liverwort Marchantia polymorpha using regenerating thalli. Biosci Biotechnol Biochem 77:167–172CrossRefGoogle Scholar
  19. 19.
    Sugano SS, Shirakawa M, Takagi J, Matsuda Y, Shimada T, Hara-Nishimura I, Kohchi T (2014) CRISPR/Cas9-mediated targeted mutagenesis in the liverwort Marchantia polymorpha L. Plant Cell Physiol 55:475–481CrossRefGoogle Scholar
  20. 20.
    Bowman JL, Kohchi T, Yamato KT, Jenkins J, Shu S, Ishizaki K, Yamaoka S, Nishihama R, Nakamura Y, Berger F, Adam C, Aki SS, Althoff F, Araki T, Arteaga-Vazquez MA, Balasubrmanian S, Barry K, Bauer D, Boehm CR, Briginshaw L, Caballero-Perez J, Catarino B, Chen F, Chiyoda S, Chovatia M, Davies KM, Delmans M, Demura T, Dierschke T, Dolan L, Dorantes-Acosta AE, Eklund DM, Florent SN, Flores-Sandoval E, Fujiyama A, Fukuzawa H, Galik B, Grimanelli D, Grimwood J, Grossniklaus U, Hamada T, Haseloff J, Hetherington AJ, Higo A, Hirakawa Y, Hundley HN, Ikeda Y, Inoue K, Inoue SI, Ishida S, Jia Q, Kakita M, Kanazawa T, Kawai Y, Kawashima T, Kennedy M, Kinose K, Kinoshita T, Kohara Y, Koide E, Komatsu K, Kopischke S, Kubo M, Kyozuka J, Lagercrantz U, Lin SS, Lindquist E, Lipzen AM, Lu CW, De Luna E, Martienssen RA, Minamino N, Mizutani M, Mochizuki N, Monte I, Mosher R, Nagasaki H, Nakagami H, Naramoto S, Nishitani K, Ohtani M, Okamoto T, Okumura M, Phillips J, Pollak B, Reinders A, Rövekamp M, Sano R, Sawa S, Schmid MW, Shirakawa M, Solano R, Spunde A, Suetsugu N, Sugano S, Sugiyama A, Sun R, Suzuki Y, Takenaka M, Takezawa D, Tomogane H, Tsuzuki M, Ueda T, Umeda M, Ward JM, Watanabe Y, Yazaki K, Yokoyama R, Yoshitake Y, Yotsui I, Zachgo S, Schmutz J (2017) Insights into land plant evolution garnered from the Marchantia polymorpha genome. Cell 171:287–304.e15CrossRefGoogle Scholar
  21. 21.
    Kubota A, Kita S, Ishizaki K, Nishihama R, Yamato KT, Kohchi T (2014) Co-option of a photoperiodic growth-phase transition system during land plant evolution. Nat Commun 5:3668CrossRefGoogle Scholar
  22. 22.
    Inoue K, Nishihama R, Kataoka H, Hosaka M, Manabe R, Nomoto M, Tada Y, Ishizaki K, Kohchi T (2016) Phytochrome signaling is mediated by phytochrome interacting factor in the liverwort Marchantia polymorpha. Plant Cell 28:1406–1421CrossRefGoogle Scholar
  23. 23.
    Kato H, Ishizaki K, Kouno M, Shirakawa M, Bowman JL, Nishihama R, Kohchi T (2015) Auxin-mediated transcriptional system with a minimal set of components is critical for morphogenesis through the life cycle in Marchantia polymorpha. PLoS Genet 11:e1005084CrossRefGoogle Scholar
  24. 24.
    Komatsu A, Terai M, Ishizaki K, Suetsugu N, Tsuboi H, Nishihama R, Yamato KT, Wada M, Kohchi T (2014) Phototropin encoded by a single-copy gene mediates chloroplast photorelocation movements in the liverwort Marchantia polymorpha. Plant Physiol 166:411–427CrossRefGoogle Scholar
  25. 25.
    Fujii Y, Tanaka H, Konno N, Ogasawara Y, Hamashima N, Tamura S, Hasegawa S, Hayasaki Y, Okajima K, Kodama Y (2017) Phototropin perceives temperature based on the lifetime of its photoactivated state. Proc Natl Acad Sci U S A 114:9206–9211CrossRefGoogle Scholar
  26. 26.
    Kodama Y (2016) Time gating of chloroplast autofluorescence allows clearer fluorescence imaging in planta. PLoS One 11:e0152484CrossRefGoogle Scholar
  27. 27.
    Suetsugu N, Takemiya A, Kong SG, Higa T, Komatsu A, Shimazaki K, Kohchi T, Wada M (2016) RPT2/NCH1 subfamily of NPH3-like proteins is essential for the chloroplast accumulation response in land plants. Proc Natl Acad Sci U S A 113:10424–10429CrossRefGoogle Scholar
  28. 28.
    Flores-Sandoval E, Eklund DM, Bowman JL (2015) A simple auxin transcriptional response system regulates multiple morphogenetic processes in the liverwort Marchantia polymorpha. PLoS Genet 11:e1005207CrossRefGoogle Scholar
  29. 29.
    Kato H, Nishihama R, Weijers D, Kohchi T (2018) Evolution of nuclear auxin signaling: lessons from genetic studies with basal land plants. J Exp Bot 69:291–301CrossRefGoogle Scholar
  30. 30.
    Okada S, Fujisawa M, Sone T, Nakayama S, Nishiyama R, Takenaka M, Yamaoka S, Sakaida M, Kono K, Takahama M, Yamato KT, Fukuzawa H, Brennicke A, Ohyama K (2000) Construction of male and female PAC genomic libraries suitable for identification of Y-chromosome-specific clones from the liverwort, Marchantia polymorpha. Plant J 24:421–428CrossRefGoogle Scholar
  31. 31.
    Ishizaki K, Chiyoda S, Yamato KT, Kohchi T (2008) Agrobacterium-mediated transformation of the haploid liverwort Marchantia polymorpha L., an emerging model for plant biology. Plant Cell Physiol 49:1084–1091CrossRefGoogle Scholar
  32. 32.
    Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158CrossRefGoogle Scholar
  33. 33.
    Takenaka M, Yamaoka S, Hanajiri T, Shimizu-Ueda Y, Yamato KT, Fukuzawa H, Ohyama K (2000) Direct transformation and plant regeneration of the haploid liverwort Marchantia polymorpha L. Transgenic Res 9:179–185CrossRefGoogle Scholar
  34. 34.
    Sugano SS, Nishihama R, Shirakawa M, Takagi J, Matsuda Y, Ishida S, Shimada T, Hara-Nishimura I, Osakabe K, Kohchi T (2018) Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLOS ONE 13:e0205117Google Scholar
  35. 35.
    Sugano SS, Nishihama R (2018) CRISPR/Cas9-based genome editing of transcription factor genes in Marchantia polymorpha. Methods Mol Biol 1830:109–126Google Scholar
  36. 36.
    Ishizaki K, Nishihama R, Yamato KT, Kohchi T (2016) Molecular genetic tools and techniques for Marchantia polymorpha research. Plant Cell Physiol 57:262–270CrossRefGoogle Scholar
  37. 37.
    Nishihama R, Ishida S, Urawa H, Kamei Y, Kohchi T (2016) Conditional gene expression/deletion systems for Marchantia polymorpha using its own heat-shock promoter and Cre/loxP-mediated site-specific recombination. Plant Cell Physiol 57:271–280CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Aino Komatsu
    • 1
  • Ryuichi Nishihama
    • 1
  • Takayuki Kohchi
    • 1
    Email author
  1. 1.Graduate School of BiostudiesKyoto UniversityKyotoJapan

Personalised recommendations