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使用Percoll梯度离心法从瓦登海沉积物中分离硅藻;硅藻产量、物种回收率和种群多样性。

Use of Percoll gradient centrifugation for the isolation of diatoms from Wadden Sea sediments; diatom yields, species recoveries and population diversity.

作者信息

Sevecke S, Morawietz T, Krumbein W E, Rhiel E

机构信息

ICBM, Carl-von-Ossietzky-Universität Oldenburg, Germany.

出版信息

Microbiologia. 1997 Jun;13(2):173-84.

PMID:9253757
Abstract

The technique of Percoll gradient centrifugation was applied to samples from Wadden Sea sediments of the shoreside near Dangast (German Bight). Experiments were also performed with Wadden Sea sediment inoculated with diatoms and with axenic and mixed diatom cultures. For axenic and mixed cultures, total recoveries ranged from 6 to 70% of the diatoms loaded onto the gradients, whereas the yields were less than 5% in most of the experiments with Wadden Sea sediments. The extraction of diatoms from mixed cultures and from sediment samples was not quantitative, neither in terms of biodiversity nor of total individuals extracted, because the rates of recovery varied for the different species. Diatom cells extracted by Percoll gradient centrifugation were devoid of sand grains and debris. The obtained material was suitable to be used in biochemical experiments, in cultural studies and for electron microscopy. The overall cell morphology and subcellular organelle organization was excellent and appeared to be unaffected by the purification procedure. The results and advantages and disadvantages of the technique are discussed.

摘要

将Percoll梯度离心技术应用于德国湾丹加斯特附近海岸边瓦登海沉积物的样本。还对接种了硅藻的瓦登海沉积物以及无菌和混合硅藻培养物进行了实验。对于无菌和混合培养物,总回收率为加载到梯度上的硅藻的6%至70%,而在大多数使用瓦登海沉积物的实验中,产量低于5%。从混合培养物和沉积物样本中提取硅藻,无论是在生物多样性方面还是在提取的个体总数方面,都不是定量的,因为不同物种的回收率不同。通过Percoll梯度离心提取的硅藻细胞没有沙粒和碎片。所获得的材料适用于生化实验、培养研究和电子显微镜观察。整体细胞形态和亚细胞器组织良好,似乎不受纯化过程的影响。讨论了该技术的结果以及优缺点。

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