Abstract
There is an increasing interest in the use of N2-fixing bacteria for the sustainable biofertilization of crops. Genetically-optimized bacteria for ammonium release have an improved biofertilization capacity. Some of these strains also cross-feed ammonium into microalgae raising additional concerns on their sustainable use in agriculture due to the potential risk of producing a higher and longer-lasting eutrophication problem than synthetic N-fertilizers. Here we studied the dynamic algal cross-feeding properties of a genetically-modified Azotobacter vinelandii strain which can be tuned to over-accumulate different levels of glutamine synthetase (GS, EC 6.3.1.20) under the control of an exogenous inducer. After switching cells overaccumulating GS into a noninducing medium, they proliferated for several generations at the expense of the previously accumulated GS. Further dilution of GS by cell division slowed-down growth, promoted ammonium-excretion and cross-fed algae. The final bacterial population, and timing and magnitude of algal N-biofertlization was finely tuned in a deferred manner. This tuning was in accordance with the intensity of the previous induction of GS accumulation in the cells. This bacterial population behavior could be maintained up to about 15 bacterial cell generations, until faster-growing and nonammonium excreting cells arose at an apparent high frequency. Further improvements of this genetic engineering strategy might help to align efficiency of N-biofertilizers and safe use in an open environment.
Key points
• Ammonium-excreting bacteria are potential eutrophication agents
• GS-dependent deferred control of bacterial growth and ammonium release
• Strong but transient ammonium cross-feeding of microalgae
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Data not included within the manuscript is available upon written request from the corresponding author.
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Acknowledgements
The authors are very thankful to Dr. Ortiz-Marquez (Boston College, MA, USA) for critical reading of the Ms. R. A. and L.C. are doctoral fellows and career researchers at the CONICET, Argentina, respectively.
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This research was funded by the Agencia Nacional de Promoción Científica y Tecnológica, Argentina [grant numbers PICT2015-3559 and PICT2018-3382] to L.C.
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L.C. conceived the project, provided funding, designed the experiments, analyzed the results, provided overall guidance, and wrote the paper. R.A. designed some experiments, executed all the experiments, analyzed the results, and helped to prepare the draft manuscript. Both authors revised the manuscript and approved it for submission.
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Ambrosio, R., Curatti, L. Deferred control of ammonium cross-feeding in a N2-fixing bacterium-microalga artificial consortium. Appl Microbiol Biotechnol 105, 2937–2950 (2021). https://doi.org/10.1007/s00253-021-11210-4
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DOI: https://doi.org/10.1007/s00253-021-11210-4