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Novel incubation‐free approaches to determine phytoplankton net primary productivity, growth, and biomass based on flow cytometry and quantification of ATP and NAD(H)
Author(s) -
Jones Bethan M.,
Halsey Kimberly H.,
Behrenfeld Michael J.
Publication year - 2017
Publication title -
limnology and oceanography: methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.898
H-Index - 72
ISSN - 1541-5856
DOI - 10.1002/lom3.10213
Subject(s) - thalassiosira pseudonana , phytoplankton , biomass (ecology) , incubation , diatom , algae , nad+ kinase , biology , botany , productivity , photosynthesis , environmental chemistry , chemistry , biochemistry , ecology , nutrient , macroeconomics , economics , enzyme
Recent advances in sorting flow cytometry (SFCM) and sensitive biochemical techniques have opened the door for a variety of new incubation‐free measurements to assess cell physiology. We adapted highly sensitive bioluminescent methodologies to measure cell NAD(H) and adenosine triphosphate (ATP) contents as predictors of phytoplankton primary production and growth rate. We first applied these methods to algae in taxonomically distinct groups: the chlorophyte Dunaliella tertiolecta , the diatom Thalassiosira pseudonana and a model cyanobacterium Synechococcus sp. WH8102. We found that NAD(H) content [NAD(H) phyto ] increased significantly with increasing net primary productivity (NPP) and growth rate. In contrast, and in agreement with previous studies, phytoplankton ATP content (ATP phyto ) was highly correlated with carbon biomass. Furthermore, ATP phyto was not correlated with NPP or growth rate. As a field test of the method, we then measured NAD(H) phyto in phytoplankton samples collected by SFCM in the North Pacific. These measurements yielded an estimate of NPP closely matching that from the traditional incubation‐dependent 14 C uptake technique. The methods described here enable low‐biomass, incubation‐free estimates of phytoplankton NPP, growth rate, and biomass.