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

研究了从两种根结线虫体内阀门中提取有机物的最佳方法。SDS-PAGE显示两种Merosira不含Slaffin，但有物质存在，CBB染色出现在3.5 kDa以下。而比都鱼的CBB条带在31-36.5 kDa之间，也在3.5 kDa以下。从直链霉菌中提取的物质通过立即与硅酸溶液混合而形成沉淀物。EDS分析表明，沉积物中含有硅和氧。沉积物由许多颗粒组成。两种直链球藻的大小不同，但在25~400 nm范围内加入磷酸后，大小发生了变化。经高效液相色谱分离纯化的物质中含有一组分，该组分与宁海因有较强的反应。用该组分进行沉淀实验，得到直径约50 nm的二氧化硅纳米球。以商品多胺为对照，显示了类似的二氧化硅纳米球，推测反应中的物质为LCPA。此外，一些其他的作用也形成了沉积物，其中一些呈网状结构，与真实的Melosira瓣非常相似。通过扫描电子显微镜观察，培养出羽状芽孢杆菌，并阐明了其瓣的发育过程。结果表明，在童虫发育完全后，其顶端融合在一起，形成了硅胶边缘环。

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.