Improved rock phosphate dissolution from organic acids is driven by nitrate assimilation of bacteria isolated from nitrate and CaCO3-rich soil

Abstract

Until now, the solubilization capacities of insoluble mineral P by soil microorganisms have been screened in vitro with media containing NH4+ as a nitrogen source. This presence of NH4+ will lead to an acidification of the medium responsible for the solubilization of the insoluble P. However, besides proton release, the production of organic acids can play a very important role in the release of free P. This physiological mechanism can largely depend on the source of nitrogen (NH4+vs NO3-) assimilated by the bacteria but the influence of the N source on the production of organic acids has yet to be studied. Our aim was to investigate if the N source assimilated by bacteria and the soil characteristics such as the dominant N source (NH4+vs NO3-) and CaCO3 contents might influence the bacterial capacities to solubilize rock phosphate. To fill this objective, we screened the capacity of bacteria isolated from 3 soils to solubilize rock phosphate in vitro in presence of NH4+or NO3-. Then, we selected the most efficient bacterial strains to identify and quantify the release of organic anions into the medium. Among the two hundred and forty-three bacterial strains isolated from the 3 soils, nine and seven isolates were identified with the highest % rock phosphate-solubilization values with NH4+ or NO3- as the sole N-source. Only one strain was able to release free Pi with NH4+ or NO3- as the sole N-source. The most predominant organic acids released by almost all isolates were gluconic acid, lactic acid, glycolic acid, acetic acid, formic acid and pyruvic acid regardless the N-source. However, with NO3- as source of N, the highest concentrations on those acids were found together with the highest release of free Pi into the medium. Molecular analysis of 16S rRNA indicated that almost all strains belonged to Bacillus and Paenibacillus genera. The PCA analysis between soil properties and bacterial capacities to release organic acids and free Pi also revealed that soil factors such as CaCO3 and soil NO3- content positively influenced the release of organic acids by bacteria grown in vitro. Our results concluded that the bacterial rock phosphate-solubilization was intimately related to organic acids production which in turn seemed to be driven by the assimilation of NO3- by bacteria. Therefore, the N-source might be considered a key factor to take into consideration during the screening and selection of suitable strains involved in the P-solubilization.

Phosphate-Solubilizing Bacteria: Advances in Their Physiology, Molecular Mechanisms and Microbial Community Effects

“…The production and secretion of organic acids are the main mechanisms of phosphorus solubilization by PSB, but the molecular mechanism of acidolysis has not been studied in depth, and there are only a few genes that can play a role in regulating phosphate solubilization (Table 6) such as gadγ (Gluconate 2-dehydrogenase), gdh (Glucose dehydrogenase), gcd (Glucose dehydrogenase) and ylil (Aldose sugar dehydrogenase) [103][104][105]. In addition to the proven gcd genes, pyruvate dehydrogenase (poxB), l-lactate dehydrogenase (ldh), glyoxylate reductase (gyaR) and glyoxylate/hydroxypyruvate/6-ketoglucuronate reductase (ghrB) are several genes associated with organic acid metabolism that may be relevant to phosphate solubilization by PSB [104].…”

“…The production and secretion of organic acids are the main mechanisms of phosphorus solubilization by PSB, but the molecular mechanism of acidolysis has not been studied in depth, and there are only a few genes that can play a role in regulating phosphate solubilization (Table 6) such as gadγ (Gluconate 2-dehydrogenase), gdh (Glucose dehydrogenase), gcd (Glucose dehydrogenase) and ylil (Aldose sugar dehydrogenase) [103][104][105]. In addition to the proven gcd genes, pyruvate dehydrogenase (poxB), l-lactate dehydrogenase (ldh), glyoxylate reductase (gyaR) and glyoxylate/hydroxypyruvate/6-ketoglucuronate reductase (ghrB) are several genes associated with organic acid metabolism that may be relevant to phosphate solubilization by PSB [104]. However, among the many genes, gluconic acid-related genes have been more intensively studied, and many Gram-negative bacteria use periplasmic glucose oxidation, mediated by pyrroloquinoline quinone (PQQ) and glucose dehydrogenase (GDH), to produce gluconic acid that encodes GCD; pyrroloquinoline quinone acts as an oxidative cofactor in glucose dehydrogenase and is mediated by the pqq manipulator genes (A, B, C, D, E, F) [2,105,106].…”