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Novel cyanobacterial bioreporters of phosphorus bioavailability based on alkaline phosphatase and phosphate transporter genes of Anabaena sp. PCC 7120

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Abstract

There is heterogeneity in the way cyanobacteria respond to P starvation and subsequently how they adapt to environments with low or fluctuating P concentrations. In this study, we have fused the promoterless lux operon luxCDABE to the promoter regions of Anabaena sp. PCC 7120 phoA genes putatively encoding alkaline phosphatases, phoA (all2843) and phoA-like (alr5291) and to the promoter region of one operon putatively encoding a high affinity phosphate transporter pst1 (all4575-4572). The self-bioluminescent strains constructed in this way, Anabaena AP (phoA promoter), Anabaena AP-L (phoA-like promoter), and Anabaena PST (pst1 promoter) have been used to study the expression of these genes in response to P starvation and P re-feeding with inorganic and organic phosphate sources. Our data showed that the pst1 promoter was activated at much higher level than the phoA-like promoter following P starvation; however, we did not observe activation of the phoA promoter. The P re-feeding experiments revealed that both strains, Anabaena (A.) PST and A. AP-L could be used as novel bioreporters of P availability in environmental samples. Both strains were used to estimate bioavailable P in environmental samples (fresh- and wastewaters) with a wide range of soluble P concentrations. The results indicated that most of the P in the water samples was in chemical forms available to the cyanobacterium; however there were some differences in the estimates given by both strains as A. PST appeared to be more adequate for the samples with the lowest P load while A. AP-L gave similar or even higher values of P concentrations than those chemically measured in samples with higher P load.

Schematic representation of the cellular elements involved in the Pho regulon in most cyanobacteria. The micrograph shows a filament of an Anabaena P-bioavailability bioreporter where P-responses are coupled to a luminescent signal

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Acknowledgments

This work was funded by Comunidad de Madrid grants S-0505/AMB/0321 and S-2009/AMB/1511 (Microambiente-CM) and by the Spanish Ministry of Science and Innovation [grant CGL2010-15675, sub-programme BOS] and [grant CGL2008-02397, sub-programme BOS]. M. Angeles Muñoz-Martín is recipient of a Post-doctoral contract from the Comunidad de Madrid. Water samples from La China STP were kindly supplied by the Confederación Hidrográfica del Tajo and those from the Guadalix River by Elvira Perona

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  1. Department of Biology, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain

    M. Ángeles Muñoz-Martín, Pilar Mateo, Francisco Leganés & Francisca Fernández-Piñas

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  1. M. Ángeles Muñoz-Martín
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  2. Pilar Mateo
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Correspondence to Francisca Fernández-Piñas.

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Muñoz-Martín, M.Á., Mateo, P., Leganés, F. et al. Novel cyanobacterial bioreporters of phosphorus bioavailability based on alkaline phosphatase and phosphate transporter genes of Anabaena sp. PCC 7120. Anal Bioanal Chem 400, 3573–3584 (2011). https://doi.org/10.1007/s00216-011-5017-0

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