ACC Deaminase Produced by PGPR Mitigates the Adverse Effect of Osmotic and Salinity Stresses in Pisum sativum through Modulating the Antioxidants Activities

Abstract

Salinity-induced ethylene production and reactive oxygen species (ROS) inhibit agricultural productivity. The plant synthesizes ethylene directly from aminocyclopropane-1-carboxylic acid (ACC). By using ACC as a nitrogen source, bacteria with ACC deaminase (ACCD) inhibit the overproduction of ethylene, thereby maintaining the ROS. The present study investigated the ACCD activity of previously identified rhizobacterial strains in Dworkin and Foster (DF) minimal salt media supplemented with 5 mM ACC (as N-source). Bacterial isolates GKP KS2_7 (Pseudomonas aeruginosa) and MBD 133 (Bacillus subtilis) could degrade ACC into α-ketobutyrate, exhibiting ACCD activity producing more than ~257 nmol of α-ketobutyrate mg protein−1 h−1, and were evaluated for other plant growth-promoting (PGP) traits including indole acetic acid production (>63 µg/mL), phosphate solubilization (>86 µg mL−1), siderophore (>20%) ammonia and exopolysaccharide production. Furthermore, Fourier Transform Infrared analysis also demonstrated α-ketobutyrate liberation from ACC deamination in DF minimal salt media, thereby confirming the ACCD activity. These isolates also showed enhanced tolerance to salinity stress of 3% w/v NaCl in vitro, in addition to facilitating multifarious PGP activities. Seed bacterization by these ACCD-producing bacterial isolates (GKP KS2_7 and MBD 133) revealed a significant decline in stress-stimulated ethylene levels and its associated growth inhibition during seedling germination. They also mitigated the negative effects of salt stress and increased the root-shoot length, fresh and dry weight of root and shoot, root-shoot biomass, total sugar, protein, reducing sugar, chlorophyll content, and antioxidants enzymes in Pisum sativum. As a result, these strains (GKP KS2_7 and MBD 133) might be applied as biofertilizers to counteract the negative effects of soil salinity.