Department of Biological Sciences, Ajou University, Suwon, 443-749, South Korea.
J Microbiol Methods. 2010 Apr;81(1):61-8. doi: 10.1016/j.mimet.2010.01.025. Epub 2010 Feb 6.
To establish molecular monitoring for the phytoplankton community in aquatic ecosystems, we analysed the terminal restriction fragment length polymorphism (T-RFLP) of small subunit ribosomal RNA gene (18S rDNA) sequences of nuclear genomes from the algal strains of culture collections and environmental samples of two freshwater reservoirs (Sangcheon reservoir and Seoho reservoir, Korea). Terminal restriction fragment (T-RF) length database was also constructed from twelve strains of algal culture collections to annotate and identify the phytoplankton species from T-RFLP profiles. Algal species in reservoirs were identified and monitored through the colony sequencing and T-RF length patterns of 18S rRNA. In this study, 41 unique clones were identified from two reservoirs including Chlorophyta, Cryptophyta, and Alveolata. In the case of Cryptomonas sp., we found significant linear relationships between T-RF peak areas and biovolumes by cell counting. Our results suggest that T-RFLP analysis can be a fast and quantitative monitoring tool for species changes in phytoplankton communities.
为了建立水生生态系统中浮游植物群落的分子监测,我们分析了藻类培养物和两个淡水水库(韩国 Sangcheon 水库和 Seoho 水库)环境样本的核基因组小亚基核糖体 RNA 基因(18S rDNA)序列的末端限制性片段长度多态性(T-RFLP)。还从 12 株藻类培养物中构建了末端限制片段(T-RF)长度数据库,以注释和鉴定 T-RFLP 图谱中的浮游植物种类。通过对 18S rRNA 的菌落测序和 T-RF 长度模式,对水库中的藻类进行鉴定和监测。在本研究中,从两个水库中鉴定出包括绿藻、隐藻和有孔虫在内的 41 个独特克隆。在 Cryptomonas sp.的情况下,我们发现 T-RF 峰面积与细胞计数的生物量之间存在显著的线性关系。我们的结果表明,T-RFLP 分析可以成为浮游植物群落物种变化的快速定量监测工具。