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Histochemical study of detailed laticifer structure and rubber biosynthesis-related protein localization in Hevea brasiliensis using spectral confocal laser scanning microscopy

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Abstract

In Hevea brasiliensis, laticifers produce and accumulate rubber particles. Despite observation using histochemical methods, development stage structure and structures with ceasing functions have rarely been described. Spectral confocal laser scanning microscopy with Nile red staining simplifies laticifer structure observation in tangential sections while enhancing the resolution. Laticifer and ray images were extracted from unmixed images and used to monitor changes during growth. A laticifer network structure developed from increased anastomoses between adjoining laticifers outside of the conducting phloem, but because of increased radial division and growth of rays, the network structure ruptured and disintegrated. We also investigated immunohistochemical localization of two rubber particle-associated proteins in the laticifers: small rubber particle protein (SRPP) and rubber elongation factor (REF). Mature bark test results show that SRPP is localized only in the laticifer layers in the conducting phloem; REF is localized in all laticifer layers. Because SRPP plays a positive role in rubber biosynthesis, results show that the rubber biosynthesis capability of laticifers is concentrated where rays and the sieve tube actively transport metabolites.

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Abbreviations

CLSM:

Confocal laser scanning microscopy

SCLSM:

Spectral confocal laser scanning microscopy

LRP:

Large rubber particle

SRP:

Small rubber particle

REF:

Rubber elongation factor

SRPP:

Small rubber particle protein

References

  • Archer BL (1980) Polyisoprene. In: Bell EA, Charlwood BV (eds) Encyclopedia of plant physiology, vol 8. Springer, Berlin, pp 309–327

    Google Scholar 

  • Archer BL, Audrey BG, Cockbain EG, McSweeney GP (1963) Biosynthesis of Rubber. Incorporation of mevalonate and isopentenyl pyrophosphate into rubber by Hevea brasiliensis latex fractions. Biochem J 89:565–574

    PubMed  CAS  Google Scholar 

  • Attanyaka DPSTG, Kekwick RGO, Franklin FCH (1991) Molecular cloning and nucleotide sequencing of the rubber elongation factor gene from Hevea brasiliensis. Plant Mol Biol 16:1079–1081

    Article  PubMed  CAS  Google Scholar 

  • Bahri ARS, Hamzah S (1996) Immunocytochemical localisation of rubber membrane protein in Hevea latex. J Nat Rubb Res 11:88–95

    CAS  Google Scholar 

  • d’Auzac J, Jacob JL, Chrestin H (1989) Physiology of rubber tree latex. CRC, Boca Raton, pp 3–55

    Google Scholar 

  • Davis W (1997) The rubber industry’s biological nightmare. Fortune 4:86–95

    Google Scholar 

  • De Bary A (1877) Vergleichende anatomie der vegetationsorgane der phanerogamen und farne. Wilhelm Engelmann, Leipzig

    Google Scholar 

  • Dickinson ME, Bearman G, Tille S, Lanford R, Fraser SE (2001) Multi-spectral imaging and linear unmixing add a whole new dimension to laser scanning fluorescence microscopy. Biotechniques 31:1272–1278

    PubMed  CAS  Google Scholar 

  • Fahn A (1979) Secretory tissues in plants. Academic, London

    Google Scholar 

  • Fowler SD, Greenspan P (1985) Application of Nile red, a fluorescent hydrophobic probe, for the detection of neutral lipid deposits in tissue sections. J Histochem Cytochem 33:833–836

    PubMed  CAS  Google Scholar 

  • Frey-Wyssling A (1930) Investigation into the relation between the diameter of the latex tubes and the rubber production of Hevea brasiliensis Arch. Voor Rubber cultuur Nederlandsch-indie 74:135–166

    Google Scholar 

  • Goyvaerts E, Dennis M, Light D, Chua NH (1991) Cloning and sequencing of the cDNA encoding the rubber elongation factor of Hevea brasiliensis. Plant Physiol 97:317–321

    Article  PubMed  CAS  Google Scholar 

  • Hamzah S, Gomez JB, Ho LH (1988) A refinement of the staining techniques for Hevea latex vessels. J Nat Rubb Res 3:163–166

    CAS  Google Scholar 

  • Hebant C, Fay E (1980) Functional organization of the bark of Hevea brasiliensis (rubber tree): a structural and histoenzymological study. Z Pflanzenphysiol 97:391–398

    Google Scholar 

  • Hebant C, Devic C, Fay E (1981) Ontogenie des laticiferes du systeme primaire de l’Hevea brasiliensis: une etude ultrastructurale et cytochimique. Can J Bot 59:974–985

    Article  Google Scholar 

  • Larson JM (2006) The Nikon C1si combines high spectral resolution, high sensitivity, and high acquisition speed. Cytometry A 69:825–834

    PubMed  Google Scholar 

  • Lynen F (1969) Biochemical problems of rubber biosynthesis. J Rubber Res Inst Malays 21:389–406

    CAS  Google Scholar 

  • Nicole M, Geiger JP, Nandris D (1986) Ultrastructure of laticifers modifications in Hevea brasiliensis infected with root rot fungi. J Phytopathol 116:259–268

    Article  Google Scholar 

  • Oh SK, Kang H, Shin DH, Yang J, Chow KS, Yeang HY, Wagner B, Breiteneder H, Han KH (1999) Isolation, characterization, and functional analysis of a novel cDNA clone encoding a small rubber particle protein from Hevea brasiliensis. J Biol Chem 274:17132–17138

    Article  PubMed  CAS  Google Scholar 

  • Ohya N, Tanaka Y, Wititsuwannakul R, Koyama T (2000) Activity of rubber transferase and rubber particle size in Hevea latex. J Rubb Res 3:214–221

    CAS  Google Scholar 

  • Omman P, Reghu CP (2003) Staining procedure for laticiferous system of Hevea brasiliensis using Oil red O. Indian J Nat Rubb Res 16:41–44

    CAS  Google Scholar 

  • Postberg J, Alexandrova O, Cremer T, Lipps HJ (2005) Exploiting nuclear duality of ciliates to analyse topological requirements for DNA replication and transcription. J Cell Sci 118:3973–3983

    Article  PubMed  CAS  Google Scholar 

  • Richardson F (1983) Technique for staining rubber in plant tissues. In: Metcalfe CR, Chalk L (eds) Anatomy of the dicotyledons, vol 2. Clarendon Press, Oxford, pp 79–80

    Google Scholar 

  • Riches JP, Gooding EGB (1952) Studies in the physiology of latex. I. Latex flow on tapping—theoretical considerations. New Phytol 51:1–10

    Article  Google Scholar 

  • Sakdapipanich JT, Suksujaritporn S, Tanaka Y (1999) Structural characterization of the small rubber particles in fresh Hevea latex. J Rubb Res 2:160–168

    CAS  Google Scholar 

  • Singh AP, Wi SG, Chung GC, Kim YS, Kang H (2003) The micromorphology and protein characterization of rubber particles in Ficus carica, Ficus benghalensis and Hevea brasiliensis. J Exp Bot 54:985–992

    Article  PubMed  CAS  Google Scholar 

  • Tan BL, Sarafis V, Beattie GAC, White R, Darley EM, Spooner-Hart R (2005) Localization and movement of mineral oil in plants by fluorescence and confocal microscopy. J Exp Bot 56:2755–2763

    Article  PubMed  CAS  Google Scholar 

  • Tarachiwin L, Sakdapipanich JT, Tanaka Y (2005) Relationship between particle size and molecular weight of rubber from Hevea brasiliensis. Rubber Chem Technol 78:694–704

    CAS  Google Scholar 

  • Wan Q, Kong L, Hao H, Wang X, Lin J, Samaj J, Baluska F (2005) Effects of brefeldin A on pollen germination and tube growth. Antagonistic effects on endocytosis and secretion. Plant Physiol 139:1692–1703

    Article  Google Scholar 

  • Wimalaratna SD (1973) A staining procedure for latex vessels of Hevea. Stain Technol 48:219–221

    PubMed  CAS  Google Scholar 

  • Wu R, Terry AV, Singh PB, Gilbert DM (2005) Differential subnuclear localization and replication timing of histone H3 lysine 9 methylation states. Mol Biol Cell 16:2872–2881

    Article  PubMed  CAS  Google Scholar 

  • Zhao XQ (1987) The significance of the structure of laticifer with relation to the exudation and latex in Hevea brasiliensis. J Nat Rubb Res 2:94–98

    Google Scholar 

Download references

Acknowledgments

Part of this work was performed in the project “Development of Fundamental Technologies for Controlling the Production of Industrial Materials by Plants”, supported by the New Energy and Industrial Technology Development Organization of Japan (NEDO). We are very grateful to Badan Pengkajian dan Penerapan Teknologi (BPPT) of Indonesia for provision of plant materials.

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Author notes

  1. Tomoki Sando

    Present address: Bridgestone Corporation, 3-1-1, Ogawahigashi, Kodaira, Tokyo, 187-8531, Japan

Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Tomoki Sando, Tatsushi Hayashi, Tsuyoshi Takeda, Yoshihisa Nakazawa, Eiichiro Fukusaki & Akio Kobayashi

  2. Bridgestone Corporation, 3-1-1, Ogawahigashi, Kodaira, Tokyo, 187-8531, Japan

    Yasunori Akiyama

Authors
  1. Tomoki Sando
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  2. Tatsushi Hayashi
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  3. Tsuyoshi Takeda
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  4. Yasunori Akiyama
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  5. Yoshihisa Nakazawa
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Correspondence to Yoshihisa Nakazawa.

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Sando, T., Hayashi, T., Takeda, T. et al. Histochemical study of detailed laticifer structure and rubber biosynthesis-related protein localization in Hevea brasiliensis using spectral confocal laser scanning microscopy. Planta 230, 215–225 (2009). https://doi.org/10.1007/s00425-009-0936-0

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  • DOI: https://doi.org/10.1007/s00425-009-0936-0

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