A novel method for the measurement of dissolved adenosine and guanosine triphosphate in aquatic habitats:: applications to marine microbial ecology

被引:30
作者
Björkman, KM [1 ]
Karl, DM [1 ]
机构
[1] Univ Hawaii, Dept Oceanog, SOEST, Honolulu, HI 96822 USA
基金
美国国家科学基金会;
关键词
ATP; GTP; microbial ecology; nucleotides;
D O I
10.1016/S0167-7012(01)00301-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel method for the measurement of dissolved adenosine-5'-triphosphate and guanosine-5'-triphosphate (D-ATP and D-GTP, respectively) in marine and freshwater habitats was developed and applied to samples collected from the oligotrophic North Pacific Ocean. Both D-ATP and D-GTP are co-precipitated by authigenically formed Mg(OH), and can be concentrated by factors greater than 200-fold, for subsequent measurement by the firefly luciferin-luciferase bioluminescence reaction. The detection limit for this method was 2-3 pmol ml(-1) of concentrated sample (equivalent to an in situ concentration of 10 pM) with a 5% precision at concentrations of 10 pmol ml(-1) or above. A significant positive correlation (P < 0.001) was observed between particulate ATP (P-ATP) and D-ATP in water samples collected from Station ALOHA (22.75 degreesN, 158 degreesW, depth profiles 0-1000 m). The highest concentrations of dissolved nucleotides were found in the euphotic zone (0-175 m) below which the concentrations were low and relatively invariant. The dissolved nucleotide pools generally exceeded their corresponding particulate pools. Using radioisotopic tracer techniques and the new concentration method, turnover times for both particulate and dissolved nucleotides can be determined. The ability to measure concentrations and follow nucleotide tracers accurately in a very dilute environment provides a unique opportunity to address questions on microbial community metabolism, nutrient dynamics and energy flux. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:159 / 167
页数:9
相关论文
共 31 条
[1]   DISSOLVED ATP IN SEA AND ITS UTILIZATION BY MARINE-BACTERIA [J].
AZAM, F ;
HODSON, RE .
NATURE, 1977, 267 (5613) :696-698
[2]   Phosphorus dynamics in the North Pacific subtropical gyre [J].
Björkman, K ;
Thomson-Bulldis, AL ;
Karl, DM .
AQUATIC MICROBIAL ECOLOGY, 2000, 22 (02) :185-198
[3]  
ECELROY WD, 1956, METHOD ENZYMOL, V2, P851
[4]   SUSCEPTIBILITY OF SOME MARINE-PHYTOPLANKTON SPECIES TO CELL BREAKAGE DURING FILTRATION AND POST-FILTRATION RINSING [J].
GOLDMAN, JC ;
DENNETT, MR .
JOURNAL OF EXPERIMENTAL MARINE BIOLOGY AND ECOLOGY, 1985, 86 (01) :47-58
[5]   MICROBIAL UPTAKE OF DISSOLVED ORGANIC-MATTER IN MCMURDO-SOUND, ANTARCTICA [J].
HODSON, RE ;
AZAM, F ;
CARLUCCI, AF ;
FUHRMAN, JA ;
KARL, DM ;
HOLMHANSEN, O .
MARINE BIOLOGY, 1981, 61 (2-3) :89-94
[6]  
HODSON RE, 1977, 2ND P ATP METH S, P127
[7]   MEASUREMENT OF ADENOSINE TRIPHOSPHATE IN OCEAN AND ITS ECOLOGICAL SIGNIFICANCE [J].
HOLMHANSEN, O ;
BOOTH, CR .
LIMNOLOGY AND OCEANOGRAPHY, 1966, 11 (04) :510-+
[8]  
Karl D.M., 1979, METHODOLOGY BIOMASS, P5
[9]  
Karl David M., 1998, P29
[10]   EFFECTS OF LUCIFERIN CONCENTRATION ON THE QUANTITATIVE ASSAY OF ATP USING CRUDE LUCIFERASE [J].
WANG, WG ;
GU, JB .
JOURNAL OF LUMINESCENCE, 1988, 40-1 :846-847