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

研究了两种直链藻（Melosira，Biddulaeae）瓣内有机物的最佳提取方法. SDS-聚丙烯酰胺凝胶电泳显示两种短链藻均不含硅亲蛋白，但在3.5kDa以下有CBB染色所见的物质。而Biddulipine显示CBB条带位于31-36.5kDa之间，也低于3.5kDa。从直链藻中提取的物质与柠檬酸溶液混合后沉淀。EDS分析表明，沉积物中含有Si和O。沉积物由许多颗粒组成。两种直链藻的大小不同，但加入磷酸后大小发生变化，为25 - 400 nm。通过HPLC纯化的物质含有显示与茚三酮强烈反应的级分。用该级分的沉降测定产生二氧化硅纳米球，约。直径50 nm。与商业产品聚胺的对照显示出类似的二氧化硅纳米球，因此认为反应物中的物质是LCPA。此外，还有一些碎屑也形成了沉积物，有些碎屑呈网状结构，与真实的直链藻的结构十分相似。结果表明，原球茎发育完全后，其顶端融合在一起，形成硅质边缘环。

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.