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Imaging the Dynamics of Cell Wall Polymer Deposition in the Unicellular Model Plant, Penium margaritaceum

  • David DomozychEmail author
  • Anna Lietz
  • Molly Patten
  • Emily Singer
  • Berke Tinaz
  • Sandra C. Raimundo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1563)

Abstract

The unicellular green alga, Penium margaritaceum, represents a novel and valuable model organism for elucidating cell wall dynamics in plants. This organism’s cell wall contains several polymers that are highly similar to those found in the primary cell walls of land plants. Penium is easily grown in laboratory culture and is effectively manipulated in various experimental protocols including microplate assays and correlative microscopy. Most importantly, Penium can be live labeled with cell wall-specific antibodies or other probes and returned to culture where specific cell wall developmental events can be monitored. Additionally, live cells can be rapidly cryo-fixed and cell wall surface microarchitecture can be observed with variable pressure scanning electron microscopy. Here, we describe the methodology for maintaining Penium for experimental cell wall enzyme studies.

Key words

Live-cell immunolabeling Penium margaritaceum Cell wall Monoclonal antibodies CLSM VP-SEM Enzymes 

Notes

Acknowledgments

This work was supported by NSF grant NSF-MCB-RUI-1517345.

References

  1. 1.
    Oikawa A, Lund CH, Sakuragi Y, Scheller HV (2013) Golgi-localized enzyme complexes for plant cell wall biosynthesis. Trends Plant Sci 18:49–58CrossRefPubMedGoogle Scholar
  2. 2.
    Li S, Lei L, Yingling YG, Gu Y (2015) Microtubules and cellulose biosynthesis: the emergence of new players. Curr Opin Plant Biol 28:76–82CrossRefPubMedGoogle Scholar
  3. 3.
    Anderson CT (2016) We be jammin’: an update on pectin biosynthesis, trafficking and dynamics. J Exp Bot 67:495–502CrossRefPubMedGoogle Scholar
  4. 4.
    Cosgrove DJ (2016) Plant cell wall extensibility: connecting plant cell growth with cell wall structure, mechanics, and the action of wall-modifying enzymes. J Exp Bot 67:463–476CrossRefPubMedGoogle Scholar
  5. 5.
    Keegstra K (2010) Plant cell walls. Plant Physiol 154:483–486CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Doblin MS, Pettolino F, Bacic A (2010) Plant cell walls: the skeleton of the plant world. Funct Plant Biol 37:357–381CrossRefGoogle Scholar
  7. 7.
    Cosgrove DJ, Jarvis MC (2012) Comparative structure and biomechanics of plant primary and secondary cell walls. Front Plant Sci3:204.http://www.frontiersin.org/Plant_Physiology/10.3389/fpls.2012.00204/full
  8. 8.
    Domozych DS, Sorensen I, Popper ZA, Ochs J, Andreas A, Fangel JU, Pielach A, Sachs C, Brechka H, Ruisi-Besares P, Willats WGT, Rose JKC (2014) Pectin metabolism and assembly in the cell wall of the charophyte green alga Penium margaritaceum. Plant Physiol 165:105–118CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Domozych DS, Sørensen I, Sacks C et al (2014) Disruption of the microtubule network alters cellulose deposition and causes major changes in pectin distribution in the cell wall of the green algaPenium margaritaceum. J Exp Bot 65:465–479CrossRefPubMedGoogle Scholar
  10. 10.
    Domozych DS, Lambiasse L, Kiemle SN, Gretz MR (2009) Cell-wall development and bipolar growth in the desmid Penium margaritaceum (Zygnematophyceae, Streptophyta). Asymmetry in a symmetric world. J Phycol 45:879–893CrossRefPubMedGoogle Scholar
  11. 11.
    Gilbert HJ, Knox JP, Boraston AB (2013) Advances in understanding the molecular basis of plant cell wall polysaccharide recognition by carbohydrate-binding modules. Curr Opin Struct Biol 23:669–677CrossRefPubMedGoogle Scholar
  12. 12.
    Mravec JJ, Kračun SK, Rydahl MG, Westereng B, Miart F, Clausen MH, Fangel JU, Daugaard M, Van Cutsem P, De Fine LHH, Höfte H, Malinovsky FG, Domozych DS, Willats WGT (2014) Tracking developmentally regulated post-synthetic processing of homogalacturonan and chitin using reciprocal oligosaccharide probes. Development 141:4841–4850CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • David Domozych
    • 1
    Email author
  • Anna Lietz
    • 1
  • Molly Patten
    • 1
  • Emily Singer
    • 1
  • Berke Tinaz
    • 1
  • Sandra C. Raimundo
    • 1
  1. 1.Department of BiologySkidmore CollegeSaratoga SpringsUSA

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