FLOW CYTOMETRIC DETERMINATION OF PHYTOPLANKTON DNA IN CULTURES AND OCEANIC POPULATIONS
FLOW CYTOMETRIC DETERMINATION OF PHYTOPLANKTON DNA IN CULTURES AND OCEANIC POPULATIONS
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DOI：
10.3354/meps071075
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发表时间：
1991-03-01
影响因子：
2.5
通讯作者：
PARTENSKY, F
中科院分区：
文献类型：
--
作者：
BOUCHER, N;VAULOT, D;PARTENSKY, F
建立了一种同时测量单个浮游植物细胞DNA和叶绿素荧光的方法，并在养殖和自然种群上进行了测试。样品用1%的戊二醛固定，在液氮中冷冻后，保存在-80℃下，用DAPI(4‘，6-二氨基-2-苯基吲哚)染色，用流式细胞仪分析。在培养物中，用流式细胞仪测量的细胞DAPI-DNA荧光与用DAPI荧光法测量的细胞DNA含量相关，几乎超过4个数量级。在自然种群中，该方法很容易将光合作用细胞与其他有生命和无生命的颗粒区分开来，并允许计算光合作用微浮游生物中含有的颗粒DNA的比例。在夏季的地中海西北部，这一比例在浅水区高于中上层站，后者在中层达到最大。在未来，这种方法将允许(1)更好地估计不同营养舱之间的生物量分配，以及(2)海洋浮游生物种群的细胞循环研究。
A method to measure DNA and chlorophyll fluorescence simultaneously in single phytoplankton cells was developed and tested on cultured and natural populations. Samples were stored at -80-degrees-C following fixation with 1% glutaraldehyde and freezing in liquid nitrogen, stained with DAPI (4',6-diamino-2-phenylindole), and analyzed by flow cytometry. In cultures, cell DAPI-DNA fluorescence measured by flow cytometry was related to cell DNA content measured by fluorometry with DAPI, over almost 4 orders of magnitude. In natural populations, the method easily discriminated photosynthetic cells from other living and non-living particles and permitted computation of the fraction of particulate DNA contained in photosynthetic picoplankton. In the northwestern Mediterranean Sea in summer, this fraction was higher at neritic than at pelagic stations, and at the latter was maximum at mid-depth. In the future, this method should permit (1) better estimates of biomass partitioning among the different trophic compartments, and (2) studies of cell cycling of oceanic plankton populations.