Flow cytometric analysis of phytoplankton viability following viral infection

被引:80
作者
Brussaard, CPD
Marie, D
Thyrhaug, R
Bratbak, G
机构
[1] Univ Bergen, Dept Microbiol, N-5020 Bergen, Norway
[2] CNRS, Biol Stn, INSU, F-29682 Roscoff, France
[3] Univ Paris 06, F-29682 Roscoff, France
关键词
flow cytometry; live/dead assays; phytoplankton; viability; viral infection;
D O I
10.3354/ame026157
中图分类号
Q14 [生态学(生物生态学)];
学科分类号
071012 ; 0713 ;
摘要
Two flow cytometric assays using physiological probes were used on the phytoplankton species Phaeocystis pouchetii and Micromonas pusilla to examine the assays' utility in detecting viral infections. Dead cells were detected using the membrane impermeant nucleic-acid dye SYTOX-Green, which stains algal cells that have lost their membrane integrity. Live cells were detected using the membrane permeant dye Calcein-AM, which is hydrolyzed by intracellular esterases into a green fluorescent charged form, We found that both assays are easy to use, are reproducible and can indeed be used as markers of the viability of individual phytoplankton cells following infection by viruses. Cell death rates up to 0.8 d(-1) for P. pouchetii and 0.5 d(-1) for Al. pusilla were calculated. The first day postinfection, death rates determined by the Calcein-AM assay were typically twice as high as those determined by the SYTOX-Green assay. Both viability tests were found to assess the physiological status of noninfected P. pouchetii cells, independent of viral infection. The optimal choice of viability assay depended on the phytoplankton species studied. Compared with existing assays, the protocols described permit examination of infected phytoplankton in more detail, yielding insight into the heterogeneity of the algal population.
引用
收藏
页码:157 / 166
页数:10
相关论文
共 32 条
[1]   Physiological stress and cell death in marine phytoplankton: Induction of proteases in response to nitrogen or light limitation [J].
Berges, JA ;
Falkowski, PG .
LIMNOLOGY AND OCEANOGRAPHY, 1998, 43 (01) :129-135
[2]   VIRUSES AS PARTNERS IN SPRING BLOOM MICROBIAL TROPHODYNAMICS [J].
BRATBAK, G ;
HELDAL, M ;
NORLAND, S ;
THINGSTAD, TF .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (05) :1400-1405
[3]   VIRAL MORTALITY OF THE MARINE ALGA EMILIANIA-HUXLEYI (HAPTOPHYCEAE) AND TERMINATION OF ALGAL BLOOMS [J].
BRATBAK, G ;
EGGE, JK ;
HELDAL, M .
MARINE ECOLOGY PROGRESS SERIES, 1993, 93 (1-2) :39-48
[4]   Autolysis kinetics of the marine diatom Ditylum brightwellii (Bacillariophyceae) under nitrogen and phosphorus limitation and starvation [J].
Brussaard, CPD ;
Noordeloos, AAM ;
Riegman, R .
JOURNAL OF PHYCOLOGY, 1997, 33 (06) :980-987
[5]   Virus-like particles in a summer bloom of Emiliania huxleyi in the North Sea [J].
Brussaard, CPD ;
Kempers, RS ;
Kop, AJ ;
Riegman, R ;
Heldal, M .
AQUATIC MICROBIAL ECOLOGY, 1996, 10 (02) :105-113
[6]   Influence of bacteria on phytoplankton cell mortality with phosphorus or nitrogen as the algal-growth-limiting nutrient [J].
Brussaard, CPD ;
Riegman, R .
AQUATIC MICROBIAL ECOLOGY, 1998, 14 (03) :271-280
[7]   Flow cytometric analyses of viral infection in two marine phytoplankton species, Micromonas pusilla (Prasinophyceae) and Phaeocystis pouchetii (Prymnesiophyceae) [J].
Brussaard, CPD ;
Thyrhaug, R ;
Marie, D ;
Bratbak, G .
JOURNAL OF PHYCOLOGY, 1999, 35 (05) :941-948
[8]   Flow cytometric detection of viruses [J].
Brussaard, CPD ;
Marie, D ;
Bratbak, G .
JOURNAL OF VIROLOGICAL METHODS, 2000, 85 (1-2) :175-182
[9]   WIDE-SPREAD OCCURRENCE AND CLONAL VARIATION IN VIRUSES WHICH CAUSE LYSIS OF A COSMOPOLITAN, EUKARYOTIC MARINE PHYTOPLANKTER, MICROMONAS-PUSILLA [J].
COTTRELL, MT ;
SUTTLE, CA .
MARINE ECOLOGY PROGRESS SERIES, 1991, 78 (01) :1-9
[10]   DYNAMICS OF A LYTIC VIRUS INFECTING THE PHOTOSYNTHETIC MARINE PICOFLAGELLATE MICROMONAS-PUSILLA [J].
COTTRELL, MT ;
SUTTLE, CA .
LIMNOLOGY AND OCEANOGRAPHY, 1995, 40 (04) :730-739