Abstract
The nitrate assimilation pathway and its regulation in the high-protein neutraceutical cyanobacterium, Arthrospira (Spirulina), were studied. A complete characterization of the genes of the nitrate uptake and assimilatory pathway in Arthrospira platensis strain PCC 7345 was done including cloning, sequencing, phylogenetic analysis and expression studies. Genomic localization studies revealed that their clustering is different from the operons known in other cyanobacteria; only nrtP and narB are organized together, while nirA, glnA and gltS exist in separate genomic locations. The presence of both types of nitrate transporters (nrtP/ABC types) in A. platensis is rare, as their occurrence is usually specific to marine and freshwater microorganisms, respectively. The positive effect of nitrate on transcript accumulation of narB, nirA and nrtP genes in N-depleted and N-restored cultures confirmed nitrate induction, which is abolished by the addition of ammonium ions into the medium. Gene expression studies in response to nitrate, nitrite, ammonium and glutamine provided the first evidence of differential regulation of multiple genes of nitrate assimilatory pathway in Arthrospira.
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Acknowledgments
We thank Prof. KJ Mukherjee of JNU, New Delhi for providing the ZAP Express Cloning kit and Dr. Arun Sharma of UDSC, New Delhi & Dr. Kiran of NRCPB (IARI), New Delhi for their initial help with the radioisotope work. This work was supported by research grant to N.R. (32-540/2006) (SR) and fellowship and research grant to S.L. from the University Grants Commission and Department of Biotechnology (BT/Bio-CARe/02/788/2010-11), New Delhi respectively.
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Communicated by Erko Stackebrandt.
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Lochab, S., Kumar, P.A. & Raghuram, N. Molecular characterization of nitrate uptake and assimilatory pathway in Arthrospira platensis reveals nitrate induction and differential regulation. Arch Microbiol 196, 385–394 (2014). https://doi.org/10.1007/s00203-014-0973-3
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DOI: https://doi.org/10.1007/s00203-014-0973-3