Isolation and characterization of phosphate-solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization

The objectives of this work were to isolate and characterize walnut phosphate-solubilizing bacteria (PSB) and to evaluate the effect of inoculation with the selected PSB stains to walnut seedlings fertilized with or without insoluble phosphate. Thirty-four PSB strains were isolated and identified under the genera Pseudomonas, Stenotrophomonas, Bacillus, Cupriavidus, Agrobacterium, Acinetobacter, Arthrobacter, Pantoea, and Rhodococcus through a comparison of the 16S ribosomal DNA sequences. All isolated PSB strains could solubilize tricalcium phosphate (TCP) in solid and liquid media. Phosphate-solubilizing activity of these strains was associated with a drop in the pH of medium. A significantly negative linear correlation was found between culture pH and phosphorus (P) solubilized from inorganic phosphate. Three isolates Pseudomonas chlororaphis (W24), Bacillus cereus (W9), and Pseudomonas fluorescens (W12) were selected for shade house assays because of their higher phosphate-solubilizing abilities. Under shade house conditions, application of W24 or W12 remarkably improved plant height, shoot and root dry weight, and P and nitrogen (N) uptake of walnut seedlings. These increases were higher on combined inoculation of PSB with TCP addition. The most pronounced beneficial effect on growth of walnut plants was observed in the co-inoculation of the three PSB strains with TCP addition. In comparison, the isolate of W9 failed to increase available soil P, nutrient levels in plants, or to promote plant growth, suggesting that more insoluble phosphate compounds than tricalcium phosphate should be used as substrates to assess the phosphate-solubilizing ability of PSB under greenhouse conditions. The present results indicated that strains P. chlororaphis or P. fluorescens could be considered for the formulation of new inoculants of walnut, even of more woody plants.