An immunological approach to detect phosphate stress in populations and single cells of photosynthetic picoplankton

被引:90
作者
Scanlan, DJ [1 ]
Silman, NJ [1 ]
Donald, KM [1 ]
Wilson, WH [1 ]
Carr, NG [1 ]
Joint, I [1 ]
Mann, NH [1 ]
机构
[1] PLYMOUTH MARINE LAB, NERC, CTR COASTAL & MARINE SCI, PLYMOUTH PL1 3DH, DEVON, ENGLAND
关键词
D O I
10.1128/AEM.63.6.2411-2420.1997
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In the marine cyanobacterium Synechococcus sp. strain WH7803, PstS is a 32-kDa cell wall-associated phosphate-binding protein specifically synthesized under conditions of restricted inorganic phosphate (P-i) availability (D. J. Scanlan, N. H. Mann, and N. G. Carr, Mol; Microbiol. 10:181-191, 1993). We have assessed its use as a potential diagnostic marker for the P status of photosynthetic picoplankton. Expression of PstS in Synechococcus sp, strain WH7803 was observed when the P-i concentration fell below 50 nM, demonstrating that the protein is induced at concentrations of P-i typical of oligotrophic conditions. PstS expression could be specifically detected by use of standard Western blotting (immunoblotting) techniques in natural mesocosm samples under conditions in which the N/P ratio was artificially manipulated to force P depletion. In addition, we have developed an immunofluorescence assay that can detect PstS expression in single Synechococcus cells both in laboratory cultures and natural samples, We show that antibodies raised against PstS cross-react with P-depleted Prochlorococcus cells, extending the use of these antibodies to both major groups of prokaryotic photosynthetic picoplankton. Furthermore, DNA sequencing of a Prochlorococcus pstS homolog demonstrated high amino acid sequence identity (77%) with the marine Synechococcus sp. strain WH7803 protein, including those residues in Escherichia coli PstS known to be directly involved in phosphate binding.
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页码:2411 / 2420
页数:10
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