Nitrogen-fixing bacteria with multiple plant growth-promoting activities enhance growth of tomato and red pepper

As a suitable alternative to chemical fertilizers, the application of plant growth-promoting rhizobacteria has been increasing in recent years due to their potential to be used as biofertilizers. In the present work, 13 nitrogen-fixing bacterial strains belonging to 11 different genera were tested for their PGP attributes. All of the strains were positive for 1-aminocyclopropane-1-carboxylate deaminase (ACCD), indole-3-acetic acid (IAA), salicylic acid, and ammonia production while negative for cellulase, pectinase, and hydrocyanic acid production. The strains Pseudomonas sp. RFNB3 and Serratia sp. RFNB14 were the most effective in solubilizing both tri-calcium phosphate and zinc oxide. In addition, all strains except Pseudomonas sp. RFNB3 were able to oxidize sulfur, and six strains were positive for siderophore synthesis. Each strain tested in this study possesses at least four PGP properties in addition to nitrogen fixation. Nine strains were selected based on their multiple PGP potential, particularly ACCD and IAA production, and evaluated for their effects on early growth of tomato and red pepper under gnotobiotic conditions. Bacterial inoculation considerably influenced root and shoot length, seedling vigor, and dry biomass of the two crop plants. Three strains that demonstrated substantial effects on plant performance were further selected for greenhouse trials with red pepper, and among them Pseudomonas sp. RFNB3 resulted in significantly higher plant height (26%) and dry biomass (28%) compared to control. The highest rate of nitrogen fixation, as determined by acetylene reduction assay, occurred in Novosphingobium sp. RFNB21 inoculated red pepper root (49.6nM of ethylene/h/g of dry root) and rhizosphere soil (41.3nM of ethylene/h/g of dry soil). Inoculation with nitrogen-fixing bacteria significantly increased chlorophyll content, and the uptake of different macro- and micro-nutrient contents enhancing also in red pepper shoots, in comparison with uninoculated controls. The population estimation studies showed that nitrogen-fixing as well as total heterotrophic bacteria were also noticeably increased in soil and plant samples. The findings of this study suggest that certain nitrogen-fixing strains possessing multiple PGP traits could be applied in the development of biofertilizers.