Abstract
Early studies in cyanobacteria have found that few genes induced by short-term salt shock (15–60 min) display a stable induction in the long-term (>1 day) salt-acclimated cells; meanwhile, most of the genes responsive to long-term salt stress were different from those by short-term salt shock, suggesting that different regulatory mechanisms may be involved for short-term and long-term salt stress responses. In our previous work using the model cyanobacterium Synechocystis sp. PCC 6803, sll1734 encoding CO2 uptake-related protein (CupA) and three genes encoding hypothetical proteins (i.e., ssr3402, slr1339, and ssr1853) were found induced significantly after a 3-day salt stress, and the corresponding gene knockout mutants were found salt sensitive. To further decipher the mechanisms that these genes may be involved, in this study, we performed a comparative metabolomic analysis of the wild-type Synechocystis and the four salt-sensitive mutants using a gas chromatography-mass spectrometry (GC-MS) approach. A metabolomic data set that consisted of 60 chemically classified metabolites was then subjected to a weighted correlation network analysis (WGCNA) to identify the metabolic modules and hub metabolites specifically related to each of the salt-stressed mutants. The results showed that two, one, zero, and two metabolic modules were identified specifically associated with the knockout events of sll1734, ssr3402, slr1339, and ssr1853, respectively. The mutant-associated modules included metabolites such as lysine and palmitic acid, suggesting that amino acid and fatty acid metabolisms are among the key protection mechanisms against long-term salt stresses in Synechocystis. The metabolomic results were further confirmed by quantitative reverse-transcription PCR analysis, which showed the upregulation of lysine and fatty acid synthesis-related genes. The study provided new insights on metabolic networks involved in long-term salt stress response in Synechocystis.
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Acknowledgments
This study was supported by funds from the National High-tech RD Program (National “863” program) (No. 2012AA02A707), the National Basic Research Program of China (National “973” program) (Nos. 2011CBA00803, 2012CB721101, and 2014CB745101), and the Tianjin Municipal Science and Technology Commission (No. 12HZGJHZ01000). The authors would also like to thank Tianjin University and the “985 Project” of the Ministry of Education for their financial support in establishing the research laboratory.
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Wang, J., Zhang, X., Shi, M. et al. Metabolomic analysis of the salt-sensitive mutants reveals changes in amino acid and fatty acid composition important to long-term salt stress in Synechocystis sp. PCC 6803. Funct Integr Genomics 14, 431–440 (2014). https://doi.org/10.1007/s10142-014-0370-7
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DOI: https://doi.org/10.1007/s10142-014-0370-7