ABSTRACT: Plant growth regulators (PGRs) and plant growth promoting rhizobacteria (PGPRs) play an important role in mitigating abiotic stresses. However, little is known about the parallel changes in physiological processes coupled with metabolic changes induced by PGRs and PGPRs that help to cope with drought stress in chickpeas. The present investigation was carried out to study the integrative effects of PGRs and PGPRs on the physiological and metabolic changes, and their association with drought tolerance in two chickpea genotypes. Inoculated seeds of two chickpea genotypes, Punjab Noor-2009 (drought sensitive) and 93127 (drought tolerance), were planted in greenhouse condition at the University of Florida. Prior to sowing, seeds of two chickpea varieties were soaked for 3 h in 24 h old cultures of PGPRs (Bacillus subtilis, Bacillus thuringiensis, and Bacillus megaterium), whereas, some of the seeds were soaked in distilled water for the same period of time and were treated as control. Plant growth regulators, salicylic acid (SA) and putrescine (Put), were applied on 25 days old seedlings just prior to the induction of drought stress. Drought stress was imposed by withholding the supply of water on 25-day-old seedlings (at the three-leaf stage) and continued for the next 25 days until the soil water content reached 14%. Ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) analysis concomitant with physiological parameters were carried out in chickpea leaves at two-time points i.e. 14 and 25 d after imposition of drought stress. The results showed that both genotypes, treated with PGRs and PGPRs (consortium), performed significantly better under drought condition through enhanced leaf relative water content (RWC), greater biomass of shoot and root, higher Fv/FM ratio and higher accumulation of protein, sugar and phenolic compounds. The sensitive genotype was more responsive than tolerant one. The results revealed that the accumulation of succinate, leucine, disaccharide, saccharic acid and glyceric acid was consistently higher in both genotypes at both time points due to PGRs and PGPRs treatment. Significant accumulation of malonate, 5-oxo-L-proline, and trans-cinnamate occurred at both time points only in the tolerant genotype following the consortium treatment. Aminoacyl-tRNA, primary and secondary metabolite biosynthesis, amino acid metabolism or synthesis pathways, and energy cycle were significantly altered due to PGRs and PGPRs treatment. It is inferred that changes in different physiological and metabolic parameters induced by PGRs and PGPRs treatment could confer drought tolerance in chickpeas.
