| 摘要: |
| 以1997年2－3月采自东海的自然海水为样品，采用SYBRGreenⅠ染色剂对微型生物细胞DNA染色后进行流式细胞仪分析，研究了SYBRGreenⅠ区分真核微型浮游植物、蓝细菌、原绿球藻以及异养细菌的效果。结果表明，该方法可较好地分离四类微型生物，同时对于一般性细胞分类和计数，可省去以前作者采用的DNA染色前处理步骤，大大提高分析速度。与传统方法相比，采用流式细胞仪对四类超微型生物进行同步监测，可避免以往不同类群生物不同方法观测所造成的系统误差，并可快速批量处理样品。 |
| 关键词: SYBRGreenⅠ 流式细胞仪 超微型生物 同步监测 |
| DOI： |
| 分类号: |
| 基金项目:国家杰出青年基金资助项目，39625008号;国家自然科学基金重点资助项目，39630060号;国家自然科学基金资助项目，39570143号 |
附件 |
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| SIMULTANEOUS MONITORING OF AUTOTROPHIC PICOPLANKTON AND HETEROTROPHIC BACTERIA |
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JIAO Nian-zhi,YANG Yan-hui
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Institute of Oceanology, The Chinese Academy of Sciences,Qingdao, 266071
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| Abstract: |
| Picoplankton is a major contributor to the total biomass and production not only in the oligotrophic oceans but also in most of the coastal waters. As different kinds of picoplankton may have different energy flow pathways and play different ecological roles in ecosyetems, detailed information on picoplankton community structure is desired for a better understanding of the function of the marine ecosystem. However, most of the data on picoplankton community structure in the China Seas were from microscopy by which an abundant tiny autotrophic picoplankton, Prochlorococcus could not be detected but miscounted as heterotrophic bacteria, leading to a significant error in biomass trophic pool budgets. Feasible analysis methods are needed for the determination of picoplankton structure in these areas. Flow cytometry (FCM) is a necessity for studying microbial ecology. The autotrophs can be easily discriminated by FCM due to their different pigment composition and cell size, while the heterotrophic bacteria can only be determined when their DNA were stained with fluorescence material. We applied flow cytometry to the simultaneous monitoring of the four principal picoplankton populations, Prochlorococcus Synechococcus, picoeukaryotes and heterotrophic bacteria in the East China Sea. We simplified the method of previous authors who used RNase, potassium citrate and Edta modifiers in pretreatment before dying and running the samples. DNA stain SYBRGreen I was added directly to a concentration of 10-4 of commercial solution 10–15 minutes before running the samples. Forward scatter, side scatter, orange fluorescence, red fluorescence and green fluorescence were used for the discrimination of these four populations. Results of the field samples by the simplified were similar to those by the previous classical method. The simple stain procedures without any pretreatment promised a feasible approach of rapid batch analysis of field picoplankton samples. |
| Key words: SYBRGreenⅠ, Flow cytometry (FCM), Picoplankton, Simultaneous monitoring |
