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J. Takahashi, S. Fujiwara, M. Kikyo, Y. Hirokawa and M. Tsuzuki, “Discrimination of the Cell Surface of the Coccolithophorid Pleurochrysis haptonemofera from Light Scattering and Fluorescein-Isothiocyanate-Labeled Lectin Staining Measured by Flow Cytometry,” Marine Biotechnology, Vol. 4, No. 1, 2002, pp. 94-101. doi:10.1007/s10126-001-0083-5

has been cited by the following article:

• TITLE: Localization and Associative Strength of Acid Polysaccharides in Coccoliths of Pleurochrysis haptonemofera (Haptophyta) Predicted from Their Extractability from Partially Decalcified Coccoliths

  AUTHORS: Yasutaka Hirokawa, Satoru Matsuzuka, Sho Itayama, Tatsuya Uchida, Shoko Fujiwara, Noriaki Ozaki, Hiromichi Nagasawa, Mikio Tsuzuki

  KEYWORDS: Acid Polysaccharide; Calcification; Coccolithophorid; Localization; Pleurochrysis haptonemofera; Urea-Extraction

  JOURNAL NAME: Open Journal of Marine Science, Vol.3 No.1, January 28, 2013

  ABSTRACT: The coccolithophorids have calcified scales called coccoliths on their surface that include abundant acid polysaccharides. To determine the localization and associative strength of acid polysaccharides Ph-PS-1, -2, and -3 incoccoliths of Pleurochrysis haptonemofera, we analyzed the acid polysaccharides extracted with urea from partially decalcified coccoliths. On treatment of coccoliths with8.0 Murea at pH8.0 inboiling water, Ph-PS-2 was not extracted, but parts of Ph-PS-1 and -3 were from the surface without the crystal morphology being affected. When coccoliths were partially decalcified at various pHs (pH 8.0 - 5.0), Ph-PS-1, -3, and -2 were extracted with urea in that order as the calcite crystals dissolved. Detection of the acid polysaccharides using fluorescein-isothiocyanate-labeled lectin (ConA) and anti-Ph-PS-2 antibodies demonstrated that Ph-PS-2 exists on the surfaces of both untreated and urea-treated coccoliths. Moreover, NaClO-treatment under moderate conditions extracted only Ph-PS-1, -3, and part of Ph-PS-2 from coccoliths, without separation of the crystal units, suggesting that at least some part of Ph-PS-2 acts as a glue connecting crystal units and/or a crystal unit and a base plate inside coccoliths. These results suggest that Ph-PS-2 exists not only on the surface but also inside of the coccoliths, most of it being strongly bound to coccoliths, while Ph-PS-1 and -3 are weakly associated close to the coccolith surface, in that order from the surface. This strongly supports a scheme in which PS-2 type acid polysaccharides play an important role in the crystal nucleation and PS-3 type functions during crystal growth.