Histochemical study of detailed laticifer structure and rubber biosynthesis-related protein localization in <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using spectral confocal laser scanning microscopy |
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Authors: | Tomoki Sando Tatsushi Hayashi Tsuyoshi Takeda Yasunori Akiyama Yoshihisa Nakazawa Eiichiro Fukusaki Akio Kobayashi |
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Institution: | (1) Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita Osaka, 565-0871, Japan;(2) Present address: Bridgestone Corporation, 3-1-1, Ogawahigashi, Kodaira Tokyo, 187-8531, Japan |
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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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Keywords: | Hevea brasiliensis Laticifer structure Rubber biosynthesis Nile red Spectral confocal laser scanning microscopy (SCLSM) Rubber particle |
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