Development of novel elongated fiber-structure in protoplast cultures of Betula platyphylia and Larix leptolepis |
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| Authors: | Sasamoto Hamako Ogita Shinjiro Hayashi Noriko Wakita Yohichi Yokota Shinso Yoshizawa Nobuo |
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| Institution: | (1) Faculty of Environment and Information, Sciences, Yokohama National University, 240-8501 Yokohama, Japan;(2) Research Association for Biotechnology, Nara Institute of Science and Technology, 630-0101 Nara, Japan;(3) Forestry and Forest Products Research Institute, 305-8687 Tsukuba, Ibaraki, Japan;(4) Greenery Research and Information Center, Hokkaido Forestry Research Institute, Bibai, 079-0198 Hokkaido, Japan;(5) Department of Forest Science, Faculty of Agriculture, Utsunomiya University, Utsunomiya, 321-8505 Tochigi, Japan |
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| Abstract: | Summary Novel elongated fiber-structures were repeatedly found both in leaf protoplast culture of two clones of Betula platyphylla and in protoplast culture of embryogenic cells of Larix leptolepis. Suboptimum culture conditions for cell division appeared to lead to fiber formation when using multi-well plate culture
with varying medium compositions The suboptimum conditions for cell divisions were brought about by (1) plant growth regulators:
auxins and cytokinins; (2) pH: 3.5, 4.5, 5.8; (3) divalent cations: CaCl2 and MgCl2; and (4) sugars: sucrose and mannitol. Divalent cations had the most profound effect on fiber formation. Calcium ions were
preferred by Betula and magnesium ions were preferred by Larix. Single fiberpurification and micro-staining methods using a micromanipulator were developed. The fibers fluoresced when
stained with Calcofluor White and Aniline Blue, which suggested that they were composed of cell wall component(s), including
callose (β-1,3-glucan). Electron microscopy showed that fiber bundles of Larix fibers had helical substructures. |
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| Keywords: | cell wall conifer fiber formation forest tree micromanipulation protoplasts |
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