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

研究了从两种木耳中提取有机物的最佳方法。SDS-PAGE显示两种Merosira不含silaffin，但有物质，CBB染色在3.5kDa位置以下出现物质。而Biddulphia显示CBB波段位于31-36.5kDa之间，也低于3.5kDa。从黄芪中获得的物质与硅酸溶液混合后立即形成沉积物。能谱分析表明，沉积物中含有硅和氧，沉积物由多个颗粒组成。两种木耳的大小不同，但随着磷酸的加入而改变，大小在25 ~ 400nm之间。高效液相色谱法纯化后的物质中含有与茚三酮反应强烈的部分。沉淀法用该组分生成直径约50nm的二氧化硅纳米球。与多胺商品化产品的对照显示相似的二氧化硅纳米球，因此在flactin中的物质应该是LCPA。此外，一些其他的弯曲也形成了沉积物，其中一些沉积物呈网状结构，与真实的Melosira阀非常相似。培养了Pennate Caloneis bacillum，并通过扫描电镜观察阐明了其瓣膜发育过程。结果表明：在微晶发育完全后，微晶顶端融合在一起，形成边缘硅环；

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.