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海洋科学  2010 

样品保藏方式与时间对海洋底栖细菌及原生生物荧光计数效能的影响

, PP. 13-20

Keywords: 冷冻,冷藏,保存时间,底栖细菌,原生生物,DAPI,荧光计数

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Abstract:

DAPI荧光染色技术已广泛应用于浮游细菌及原生生物的定量研究,但对底栖生物的定量效能尚缺必要的研究。比较了冷冻和冷藏两种保藏方式以及保存时间对DAPI荧光计数底栖细菌、蓝细菌、硅藻、不同粒级自养小鞭毛虫(PNF)和异养小鞭毛虫(HNF)的影响。对黄海冷水团三个站位表层2cm底栖样品进行的4℃冷藏与?20℃冷冻避光保存的比较研究表明,两种保藏方式下两个站位的所有研究对象均无显著差异,但另一站位的PNF(2~5μm和5~10μm)的冷藏保存显著优于冷冻保存。对选取的另外两个站位(0~2cm和2~5cm分层)的样品经1个月和4个月冷藏保存后的分析表明,对于底栖细菌、蓝细菌、PNF(5~10μm)、PNF(>10μm)、HNF(>10μm)和硅藻在保存1个月和4个月后的计数没有显著差异,而对于PNF(2~5μm)、HNF(2~5μm)、HNF(5~10μm)保存4个月的数量明显低于保存1个月的样品,如其中一个站位的0~2cm分层的PNF(2~5μm)丰度减少了47.4%,2~5cm分层的丰度减少了59.6%,HNF(2~5μm)和HNF(5~10μm)经4个月后丰度降为0。本研究表明,对底栖细菌、蓝细菌以及原生生物的定量计数可因样品、保藏方式及保存时间的不同而产生差异,因此对于底栖样品短期内宜采用避光、冷藏保存,并在带回实验室后尽快分析。

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