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Imaging of early conifer embryogenic tissues with the environmental scanning electron microscope

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Biologia Plantarum

Abstract

This article describes the usage of non-commercial environmental scanning electron microscope (ESEM) for the visualization of plant extracellular matrix in Abies alba and Abies numidica. Non sputter-coated samples free of using the common fixation technique observed at the relatively low humidity of the air environment with the pressure 550 Pa and the low temperature of the sample from −18 to −22°C give surprisingly very good results that show the natural structure of the tissues. This seems to be generally applicable. Moreover, a specially designed ionization detector of secondary electrons and a YAG:Ce3+ detector of backscattered electrons were used for better comparison.

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Abbreviations

SH medium:

Schenk and Hildebrandt medium

AGPs:

arabinogalactan proteins

ECM:

extracellular matrix

ECMSN:

extracellular matrix surface network

SEM:

scanning electron microscope

ESEM:

environmental SEM

SE:

secondary electrons

BSE:

back secondary electrons

YAG: Ce3+ :

yttrium aluminum garnet activated with trivalent cerium

References

  • Bobák, M., Šamaj, J., Hlinková E., Hlavačka A., Ovečka M.: Extracellular matrix in early stages of direct somatic embryogenesis in leaves of Drosera spathulata. — Biol. Plant. 47: 161–166, 2003/4.

    Article  Google Scholar 

  • Danilatos, G.D.: Foundations of environmental scanning electron microscopy. — Adv. Electronics Electron Phys. 71: 109–250, 1988.

    Article  CAS  Google Scholar 

  • Dubois, T., Dubois, J., Guedira, M., Diop, A., Vasseur, J.: SEM characterization of an extracellular matrix around somatic proembryos in roots of Cichorium. — Ann. Bot. 70: 119–124, 1992.

    Google Scholar 

  • Hakman, I., Fowke, L.C.: Somatic embryogenesis in Picea glauca (white spruce) and Picea mariana (black spruce). — Can. J. Bot. 65: 656–659, 1987.

    Article  Google Scholar 

  • Hřib, J.: Study of somatic and zygotic embryos of Algerian fir (Abies numidica De Lann. Carrière) by AQUASEM. — In: Libiaková, G., Gajdošová, A. (ed.): Book of Abstract COST 843 Final Conference, COST 843 and COST 851 Joint Meeting. Pp. 269–271. Institute of Plant Genetics and Biotechnology SAS, Stará Lesná 2005.

    Google Scholar 

  • Jalonen, P., Von Arnold, S.: Characterization of embryogenic cell lines of Picea abies in relation to their competence for maturation. — Plant Cell Rep. 10: 384–387, 1991.

    Article  Google Scholar 

  • Mo, L.H., Egertsdotter, U., Von Arnold, S.: Secretion of specific extracellular proteins by somatic embryos of Picea abies is dependent on embryo morphology. — Ann. Bot. 77: 143–152, 1996.

    Article  CAS  Google Scholar 

  • Neděla, V.: Methods for additive hydratation allowing observation of fully hydrated state of wet samples in environmental SEM. — Microscope Res. Tech. 70: 95–100, 2007.

    Article  Google Scholar 

  • Neděla, V.: Controlled dehydration of a biological sample using an alternative form of environmental SEM. — J. Microscopy 237: 7–11, 2010.

    Article  Google Scholar 

  • Overvoode, P.J., Grimes, H.D.: The role of calcium and calmodulin in carrot somatic embryogenesis. — Plant Cell Physiol. 32: 135–144, 1994.

    Google Scholar 

  • Popielarska-Konieczna, M., Bohdanowicz, J., Starnawska, E.: Extracellular matrix of plant callus tissue visualized by ESEM and SEM. — Protoplasma 247: 121–125, 2010.

    Article  PubMed  Google Scholar 

  • Reid, D.A., Lott, J.N.A., Attree, S.M., Fowke, L.C.: Imbibition of white spruce seeds and somatic embryos: a study of morphological changes in an environmental scanning electron microscope and potassium leakage. — In Vitro cell. dev. Biol. Plant. 35: 303–308, 1999.

    Article  CAS  Google Scholar 

  • Ruoslahti, E., Pierschbacher, M.D.: New perspectives in cell adhesion: RGD and integrins. — Science 238:491–497, 1987.

    Article  PubMed  CAS  Google Scholar 

  • Šamaj, J., Bobák, M., Blehová, A., Krištin, J., AuxtováŠamajová, O.: Developmental SEM observations on an extracellular matrix in embryogenic calli of Drosera rotundifolia and Zea mays. — Protoplasma 186: 45–49, 1995.

    Article  Google Scholar 

  • Šamaj, J., Salaj, T., Matúšová, R., Salaj, J., Takáč, T., Šamajová, O., Volkmann, D.: Arabinogalactan-protein epitope Ga14 is differentially regulated and localized in cell lines of hybrid fir (Abies alba × Abies cephalonica) with different embryogenic and regeneration potential. — Plant Cell Rep. 27: 221–229, 2008.

    Article  PubMed  Google Scholar 

  • Schenk, R.U., Hildebrandt, A.C.: Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. — Can. J. Bot. 50: 199–204, 1972.

    Article  CAS  Google Scholar 

  • Sondahl, M.R., Salisbury, J.L., Sharp, W.R.: SEM characterization of embryogenic tissue and globular embryos during high frequency somatic embryogenesis in coffee callus cells. — Z. Pflanzenphysiol. 94: 185–188, 1979.

    Google Scholar 

  • Stokes, D.J.: Investigating biological ultrastructure using environmental scanning electron microscopy (ESEM). — In: Mendez-Vilas, A. (ed.): Science, Technology and Education of Microscopy: an Overview. Vol. 1. Pp. 564–570. Formatex Publications, Badajoz 2003.

    Google Scholar 

  • Timmers, A.C.J., De Vries. S.C., Schel, J.H.N.: Distribution of membrane-bound calcium and activated calmodulin during somatic embryogenesis of carrot (Daucus carota L.). — Protoplasma 153: 24–29, 1989.

    Article  Google Scholar 

  • Vooková, B., Kormuťák, A.: Improved plantlet regeneration from open-pollinated families of Abies alba trees of Dobroč primeval forest and adjoing managed stand via somatic embryogenesis. — Biologia (Bratislava) 64: 1136–1140, 2009.

    Article  Google Scholar 

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Authors and Affiliations

  1. Institute of Scientific Instruments, Academy of Sciences of the Czech Republic, Královopolská 147, CZ-61264, Brno, Czechoslovakia

    V. Neděla

  2. Institute of Scientific Instruments, Academy of Sciences of the Czech Republic, Ukrajinská 17, CZ-62500, Brno, Czechoslovakia

    J. Hřib

  3. Czech Republic Institute of Plant Genetics and Biotechnology, SAS, Akademická 2, SK-95007, Nitra, Slovak Republic

    B. Vooková

Authors
  1. V. Neděla
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  2. J. Hřib
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  3. B. Vooková
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Correspondence to V. Neděla.

Additional information

Acknowledgements: This work was supported by the Grant Agency of the Czech Republic, grant No. GAP 102/10/1410 and the Slovak Grant Agency for Science, Project No. 2/0076/09.

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Neděla, V., Hřib, J. & Vooková, B. Imaging of early conifer embryogenic tissues with the environmental scanning electron microscope. Biol Plant 56, 595–598 (2012). https://doi.org/10.1007/s10535-012-0062-x

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  • DOI: https://doi.org/10.1007/s10535-012-0062-x

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