Thiamethoxam, a broad spectrum, neonicotinoid insecticide, is used on various crops including Brassica juncea L. to protect from intruding insects such as leaf-hoppers, aphids, thrips and white-flies. Exposure to thiamethoxam causes acute malady such as tumour development, cell apoptosis, liver damage and neurotoxicity. Melatonin is entailed in umpteen developmental processes of plants, including stress responses. The pleiotropic effects of melatonin in modulating plant growth validate it's imperative contribution as multi-regulatory substance. Exiguous information is known about the role of Pseudomonas putida in improving plant growth under thiamethoxam stress. Taking these aspects into consideration the contemporary study investigates the role of melatonin and Pseudomonas putida strain MTCC 3315 in alleviating the thiamethoxam induced toxicity in B. juncea plant. Fourier Transform Infrared Spectroscopy (FTIR) analysis uncloaked that thiamethoxam induced stress primarily affects the protein content of plant as compared to lipids, carbohydrates and cell wall components. Organic acid profiling of the treated samples carried-out by High-Performance Liquid Chromatography (HPLC), reported an upregulation in the level of organic acids, malic acid (110%), citric acid (170%), succinic acid (81%), fumaric acid (40%) and ascorbic acid (55%) in thiamethoxam treated plants compared to the investigational untreated plants. The melatonin treated seedlings grown under thiamethoxam stress, exhibit increased level of malic acid, citric acid, succinic acid, fumaric acid and ascorbic acid by 81%, 0.94%, 11%, 21% and 6% respectively. Further, thiamethoxam stressed plants inoculated with Pseudomonas putida showed stupendous up-regulation by 161% (malic acid), by 14% (citric acid), by 33% (succinic acid), by 30% (fumaric acid), by 100% (oxalic acid) respectively. Lastly, the combinatorial application of melatonin and Pseudomonas putida resulted in prodigious upsurge of malic acid by 165%, succinic acid by 69%, fumaric acid by 42% respectively in contrast to distinct melatonin and Pseudomonas putida treatments. The accumulation of organic acids ascertains the defence against thiamethoxam stress and corresponds to meet the energy generation requirement to skirmish thiamethoxam mediated abiotic stress in Brassica juncea plant.
