Skip to main content
SpringerLink
Log in

Atomic force microscopy and laser confocal scanning microscopy analysis of callose fibers developed from protoplasts of embryogenic cells of a conifer

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Efficiency of novel fiber formation was much improved in protoplast culture of embryogenic cells (ECs) of a conifer, Larix leptolepis (Sieb. et Zucc.) Gord., by pre-culturing ECs in a medium containing a high concentration of glutamine (13.7 mM). The fibrillar substructures of large and elongated fibers of protoplasts isolated from Larix ECs were investigated by laser confocal scanning microscopy (LCSM) after Aniline Blue staining and atomic force microscopy (AFM) using a micromanipulator without any pre-treatment. Fibers were composed of bundles of fibrils and subfibrils, whose diameters were defined as 0.7 and 0.17 μm, respectively, by image analysis after LCSM and AFM. These fibers were proven to be composed of callose by using specific degrading enzymes for β-1,4-glucan and β-1,3-glucan.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Cheadle VI, Gifford EM, Esau K (1953) A staining technique for phloem and contiguous tissue. Stain Technol 28:49–53

    PubMed  CAS  Google Scholar 

  • Fukumoto T, Sasamoto H, Hayashi N, Ogita S, Wakita Y, Yokota S, Yoshizawa N (2004) Development of novel large elongated fiber-structure in plant protoplast cultures. In Vitro Cell Dev Biol 40:73A

    Google Scholar 

  • Hahne G, Herth W, Hoffmann F (1983) Wall formation and cell division in fluorescence-labelled plant protoplasts. Protoplasma 115:217–221

    Article  Google Scholar 

  • Hayashi N, Sugiyama J, Okano T, Ishihara M, Shimizu K (1998a) Selective degradation of cellulose Ia component in Cladophora cellulose with Trichoderma viride cellulase. Carbohydrate Res 305:109–116

    Article  Google Scholar 

  • Hayashi N, Sugiyama J, Okano T, Ishihara M, Shimizu K (1998b) The enzymatic susceptibility of cellulose microfibrils of the algal-bacterial type and the cotton-ramie type. Carbohydrate Res 305:261–269

    Article  Google Scholar 

  • Hoffmann F (1996) Laser microbeams for the manipulation of plant cells and subcellular structures. Plant Sci 113:1–11

    Article  CAS  Google Scholar 

  • Murashige T and Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Ogita S, Kubo T, Fushitani M (1997) Anatomical characteristics in early embryogenesis from immature embryo of Larix leptolepis. For Res Environ 35:45–51

    Google Scholar 

  • Ogita S, Sasamoto H, Kubo T (1999) Selection and microculture of single embryogenic cell clusters in Japanese conifers: Picea jezoensis, Larix leptolepis and Cryptomeria japonica. In Vitro Cell Dev Biol Plant 35:428–431

    Article  Google Scholar 

  • Ogita S, Sasamoto H, Yeung EC, Thope TA (2001) The effects of glutamine on the maintenance of embryogenic cultures of Cryptomeria japonica. In Vitro Cell Dev Biol Plant 37:268–273

    Article  CAS  Google Scholar 

  • Pesacreta TC, Carlson LC, Triplett BA (1997) Atomic force microscopy of cotton fiber cell wall surfaces in air and water: quantitative and qualitative aspects. Planta 202:435–442

    Article  CAS  Google Scholar 

  • Sasamoto H, Ogita S, Yeung, EC, Thorpe TA (2001) Protoplast culture of Larix kaempferi and Cryptomeria japonica: Regulatory factors for protoplast fiber formation and early somatic embryogenesis (in Japanese). Trans Jpn For Soc (ISSN 1341–6960) 112:660

  • Sasamoto H, Ogita S, Fukui M (2002) Cell engineering of Japanese conifers, Cryptomeria japonica and Larix kaempferi. In Vitro Cell Dev Biol 38: p56A

    Google Scholar 

  • Sasamoto H, Ogita S, Hayashi N, Wakita Y, Yokota S, Yoshizawa N (2003) Development of novel elongated fiber-structure in protoplast cultures of Betula platyphylla and Larix leptolepis. In Vitro Cell Dev Biol Plant 39:223–228

    Article  Google Scholar 

  • Yeung EC (1984) Histological and histochemical staining procedures. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants, vol. 1. Academic Press, Orlando, FL, pp 689–697

    Google Scholar 

Download references

Acknowledgements

Thanks are due to Drs. A. Takebe and N. Oba for their kind help with image analysis using LCSM. T.F. is a research fellow of the Japan Society for the Promotion of Science (JSPS) for Young Scientists.

Author information

Authors and Affiliations

  1. Faculty of Environment and Information Sciences, Yokohama National University, Yokohama, 240-8501, Kanagawa, Japan

    Takeshi Fukumoto & Hamako Sasamoto

  2. Forestry and Forest Products Research Institute, Tsukuba, 305-8687, Ibaraki, Japan

    Noriko Hayashi

Authors
  1. Takeshi Fukumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Noriko Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hamako Sasamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hamako Sasamoto.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fukumoto, T., Hayashi, N. & Sasamoto, H. Atomic force microscopy and laser confocal scanning microscopy analysis of callose fibers developed from protoplasts of embryogenic cells of a conifer. Planta 223, 40–45 (2005). https://doi.org/10.1007/s00425-005-0065-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-005-0065-3

Keywords

Search

Navigation