Engineering of a photosynthetic diazotroph as biofertilizer for efficient nitrogen fixation powered by sunlight

Abstract

Background Photosynthetic diazotrophs expressing iron-only (Fe-only) nitrogenase can be developed into a promising biofertilizer, as it can accomplish the energy-intensive process of nitrogen fixation by light energy and is independent on the molybdenum availability in the soil. However, the expression of Fe-only nitrogenase in diazotrophs is repressed by the fixed nitrogen of the soil, limiting the efficiency of nitrogen fixation in farmland with low ammonium concentrations that are inadequate for sustainable crop growth. Results We succeeded in constitutively expressing the Fe-only nitrogenase even in the presence of ammonium by controlling the transcription of Fe-only nitrogenase gene cluster (anfHDGK) with the transcriptional activator of Mo nitrogenase (NifA*) in several different ways, indicating that the engineered NifA* strains can be used as promising chassis cells for efficient expression of different types of nitrogenases. When applied as a biofertilizer, the engineered Rhodopseudomonas palustris effectively stimulated rice growth, contributing to the reduced use of chemical fertilizer and the development of sustainable agriculture. Conclusion We developed the engineered R. palustris strains that constitutively express Fe-only nitrogenase in the presence of ammonium for the first time. These R. palustris strains driven by sunlight can be used as efficient nitrogen biofertilizers.