Localization of sucrose synthase and callose in freeze-substituted secondary-wall-stage cotton fibers |
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Authors: | Salnikov Vadim V Grimson Mark J Seagull Robert W Haigler Candace H |
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Institution: | (1) Department of Biological Sciences, Texas Tech University, Lubbock, Texas, US;(2) Department of Biology, Hofstra University, Hempstead, New Tork, US |
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Abstract: | Summary. Methods for cryogenic fixation, freeze substitution, and embedding were developed to preserve the cellular structure and
protein localization of secondary-wall-stage cotton (Gossypium hirsutum L.) fibers accurately for the first time. Perturbation by specimen handling was minimized by freezing fibers still attached
to a seed fragment within 2 min after removal of seeds from a boll still attached to the plant. These methods revealed native
ultrastructure, including numerous active Golgi bodies, multivesicular bodies, and proplastids. Immunolocalization in the
context of accurate structure was accomplished after freeze substitution in acetone only. Quantitation of immunolabeling identified
sucrose synthase both near the cortical microtubules and plasma membrane and in a proximal exoplasmic zone about 0.2 μm thick.
Immunolabeling also showed that callose (β-1,3-glucan) was codistributed with sucrose synthase within this exoplasmic zone.
Similar results were obtained from cultured cotton fibers. The distribution of sucrose synthase is consistent with its having
a dual role in cellulose and callose synthesis in secondary-wall-stage cotton fibers.
Received August 19, 2002; accepted November 12, 2002; published online June 13, 2003
RID="*"
ID="*" Correspondence and reprints: Department of Biological Sciences, Texas Tech University, Lubbock, TX 79409-3131, U.S.A.
E-mail: candace.haigler@ttu.edu |
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Keywords: | : Cell wall Cellulose synthesis Cryogenic specimen preparation Gossypium hirsutum Immunolabeling Transmission electron microscopy |
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