海洋甲藻的单细胞PCR技术初步研究

甲藻是引发赤潮的重要种类。但是由于某些种类个体微小、形态近似,很难通过光学形态特征准确鉴定。单细胞PCR是一种快速、有效的分子鉴定方法。分别用戊二醛、鲁哥氏液和福尔马林固定塔玛亚历山大藻(Alexandrium tarmarense)。塔玛亚历山大藻单细胞全基因组扩增后进行单细胞PCR扩增,结果显示:1个月内,2%戊二醛和鲁格氏液固定甲藻均能用于单细胞PCR分析,而福尔马林固定甲藻则不适宜单细胞PCR分析。同时,2%戊二醛固定大连凌水湾海域甲藻样品,结合光学形态特征分析和单细胞PCR,成功鉴定了环境样品中的锥形多甲藻。

The preliminary study of single-cell PCR analysis of marine Dinoflagellates

Dinoflagellates is one of the important harmful algae bloom forming species.Because of the similarity in size and shape,micro-dinoflagellates were difficult to be identified exactly using morphological approaches.Single-cell PCR is probably the most rapid and convenient method that can identify marine dinoflagellates accurately.In this study,Alexandrium tarmarense were fixed with glutaraldehyde,Lugol's solution and formalin respectively.Before undertaken single-cell PCR,the whole genome of single cell was amplified.The results showed that the cells preserved in glutaraldehyde and Lugol's solution could be analyzed using sing-cell PCR in 30 days.While there were negative results in the A.tarmarense cells preserved in formalin.The dinoflagellate Peridinium conicum collected in Lingshui Bay in Dalian were fixed with glutaraldehyde at final concentration of 2%,and successfully identified by combining light microscopy and single cell PCR Amplification methods.