To colonize the cystic fibrosis lung, Pseudomonas aeruginosa establishes sessile communities referred to as biofilms. Although the signaling molecule c-di-GMP governs the transition from motile to sessile growth, the environmental signal(s) required to modulate biofilm formation remain unclear. Using relevant in vivo concentrations of the 19 amino acids previously identified in cystic fibrosis sputum, we demonstrated that arginine, ornithine, isoleucine, leucine, valine, phenylalanine and tyrosine robustly promoted biofilm formation in vitro. Among the seven biofilm-promoting amino acids, only arginine also completely repressed the ability of P. aeruginosa to swarm over semi-solid surfaces, suggesting that arginine may be an environmental cue favoring a sessile lifestyle. Mutating two documented diguanylate cyclases required for biofilm formation (SadC and RoeA) reduced biofilm formation and restored swarming motility on arginine-containing medium. Growth on arginine increased the intracellular levels of c-di-GMP, and this increase was dependent on the SadC and RoeA diguanylate cyclases. Strains mutated in sadC, roeA or both also showed a reduction in biofilm formation when grown with the other biofilm-promoting amino acids. Taken together, these results suggest that amino acids can modulate biofilm formation and swarming motility, at least in part, by controlling the intracellular levels of c-di-GMP.

This research was designed to elucidate the role of exopolysaccharides (EPS) producing bacterial strains for the amelioration of drought stress in wheat. Bacterial strains were isolated from a farmer’s field in the arid region of Pakistan. Out of 24 isolated stains, two bacterial strains, Bacillus subtilis (Accession No. MT742976) and Azospirillum brasilense (Accession No. MT742977) were selected, based on their ability to produce EPS and withstand drought stress. Both bacterial strains produced a good amount of EPS and osmolytes and exhibited drought tolerance individually, however, a combination of these strains produced higher amounts of EPS (sugar 6976 µg/g, 731.5 µg/g protein, and 1.1 mg/g uronic acid) and osmolytes (proline 4.4 µg/mg and sugar 79 µg/mg) and significantly changed the level of stress-induced phytohormones (61%, 49% and 30% decrease in Indole Acetic Acid (IAA), Gibberellic Acid (GA), and Cytokinin (CK)) respectively under stress, but an increase of 27.3% in Abscisic acid (ABA) concentration was observed. When inoculated, the combination of these strains improved seed germination, seedling vigor index, and promptness index by 18.2%, 23.7%, and 61.5% respectively under osmotic stress (20% polyethylene glycol, PEG6000). They also promoted plant growth in a pot experiment with an increase of 42.9%, 29.8%, and 33.7% in shoot length, root length, and leaf area, respectively. Physiological attributes of plants were also improved by bacterial inoculation showing an increase of 39.8%, 61.5%, and 45% in chlorophyll a, chlorophyll b, and carotenoid content respectively, as compared to control. Inoculations of bacterial strains also increased the production of osmolytes such asproline, amino acid, sugar, and protein by 30%, 23%, 68%, and 21.7% respectively. Co-inoculation of these strains enhanced the production of antioxidant enzymes such as superoxide dismutase (SOD) by 35.1%, catalase (CAT) by 77.4%, and peroxidase (POD) by 40.7%. Findings of the present research demonstrated that EPS, osmolyte, stress hormones, and antioxidant enzyme-producing bacterial strains impart drought tolerance in wheat and improve its growth, morphological attributes, physiological parameters, osmolytes production, and increase antioxidant enzymes.

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