Phylogenetic study of diatoms based on kinetic morphology formed during valve ontogeny and molecular associated with bio-silica mineralization

基于瓣膜个体发育过程中形成的动力学形态和与生物硅矿化相关的分子的硅藻系统发育研究

基本信息

批准号：
15570075
负责人：
MAYAMA Shigeki
金额：
$ 2.37万
依托单位：
Tokyo Gakugei University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Optimal method for extructing organic matters from valve inside of two Melosira, Biddulphia was studied. SDS-PAGE showed that two Merosira species did not include silaffin but had matter, which appeared by CBB staining below the popsition of 3.5kDa. Whereas Biddulphia showed CBB band positioned between 31-36.5kDa and also below 3.5kDa. The matter obtained from Melosira make sediment by mixing with silicic acid solution imediately. EDS analysis indicated that the sediment matter containig Si and O. The sediment was compposed of many granula. The sizes were different between two Melosira species, but it changed by addition of phosphate acid, 25 to 400nm. Pureified matter by HPLC contained a fraction which showed strong reaction with Ninhydryn. Sedimentation assay with this fraction generated silica-nanosphere, ca. 50nm in diameter. Control with commercial product of polyamin showed similar silica-nanosphere, so that the matter in the flaction was supposed to be LCPA. In addition, some othere flaction also made sediments, and some of them showed mesh-like structure, which was quite similar to that of real Melosira valve.Pennate Caloneis bacillum was cultered and proceedure of its valve development was clarified by SEM observations. The result showed that after complete development of virgae, their apices were fused together and marginal ring of silica was formed.

研究了两种Melosira、Biddulphia瓣膜内有机物提取的最佳方法。 SDS-PAGE显示，两种Merosira属不含有硅蜡，但有物质，CBB染色显示其分子量低于3.5kDa。而 Biddulphia 显示 CBB 带位于 31-36.5kDa 之间，也低于 3.5kDa。从Melosira获得的物质立即与硅酸溶液混合形成沉淀。 EDS分析表明沉积物中含有Si和O。沉积物由许多颗粒组成。两种Melosira 物种之间的尺寸不同，但通过添加磷酸而改变，25 至400nm。通过HPLC纯化的物质含有显示出与茚三酮强烈反应的级分。用该部分进行沉降测定产生二氧化硅纳米球，约。直径50纳米。用市售产品聚胺进行对照，显示出类似的二氧化硅纳米球，因此馏分中的物质被认为是LCPA。此外，其他一些部分也产生沉积物，其中一些呈网状结构，与真实的Melosira瓣膜非常相似。对Pennate Caloneis bacillum进行了培养，并通过SEM观察阐明了其瓣膜的发育过程。结果表明，菖蒲发育完全后，其顶端融合在一起，并形成二氧化硅边缘环。