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四类海洋超微型浮游生物的同步监测
SIMULTANEOUS MONITORING OF AUTOTROPHIC PICOPLANKTON AND HETEROTROPHIC BACTERIA
【摘要】 以1997年2-3月采自东海的自然海水为样品,采用SYBRGeenⅠ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SYBRGeenⅠ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。
【Abstract】 Picoplankton is a major contributor to the total biomass and production not only in theoligotrophic oceans but also in most of the coastal waters. As different kinds of Picoplankton mayhave different energy flow pathways and play different ecological roles in ecosystems, detailedinformation on picoplankton community structure is desired for a better understanding of the functionof the marine ecosystem. However, most of the data on picoplankton community structurein theChina Seas were from microscopy by which an abundant tiny autotrophjc picoplankton,Prochlorococcus could not be detected but miscounted as heterotrophic bacteria, leading to asighificant error in biomass trophic pool budgets. Feasible analysis methods are needed for thedeterminahon of picoplankton structure in these areas. Flow cytomeny (FCM)is a necessity forstudying microbial ecology. The autotrophs can be easily discriminated by FCM due to theirdifferent pigment composition and cell size, while the heterotrophjc bacteria can only be determinedwhen their DNA were stained with fluorescence material. We applied flow cytometry to thesimultaneous monitoring of the four principal picoplankton Populahons, Prochlorococcus,Synechococcus, picoeukaryotes and heterotrophic bacteria in the East China Sea. We simplified themethod of previous authors who used RNase, Potassitum citrate and EDTA modifiers in pretreatmentbe fore dying and running the samples. DNA snin SYBRGreen I was added direchy to aconcentration of 10 -4 of commercial solution 10- 15 minutes before running the samples. Forwardscader, side scatter, orange fluorescence, red fluorescence and green fluorescence were used for thediscrimination of these four populations. Results of the field samples by the simplified were similarto those by the previous classical method. The simple strin procedures without any preteatmentpromised a feasible approach of rapid batch analysis of field picoplankton samples.
【Key words】 SYBRGreen Ⅰ Flow cytometry (FCM) Picoplankton Simultaneous monitoring;
- 【文献出处】 海洋与湖沼 ,OCEANOLOGIA ET LIMNOLOGIA SINICA , 编辑部邮箱 ,1999年05期
- 【分类号】Q-331
- 【被引频次】76
- 【下载频次】496