Melatonin-mediated methylglyoxal homeostasis and regulation of autophagy during seed germination under PEG-induced drought stress in upland cotton

Methylglyoxal (MGO), toxic byproduct of glycolysis, acts as a signaling molecule at low levels, but its over-accumulation during drought stress disrupts redox balance and accelerates cell death. Contrarily, melatonin maintains redox balance, particularly during stress. The redox status and MGO level might differ in drought-sensitive and drought-tolerant varieties, so shall the melatonins effect. The study evaluated the effect of melatonin-priming on MGO detoxification and autophagy during polyethylene glycol (PEG)-induced drought stress during seed germination in drought-sensitive (L-799) and drought-tolerant (Suraj) varieties of upland cotton. Melatonin-priming increased endogenous melatonin content, reduced MGO accumulation and advanced glycation end-products (AGEs), and downregulated the expression of MGO biosynthesis genes in L-799 under stress. The expression and activities of glyoxalases and non-glyoxalases were upregulated, showing melatonins effectiveness in MGO detoxification. Additionally, priming upregulated the expression of TPI1, PGK5, and PK1 and downregulated HK3 expression, allowing better conversion of glucose to pyruvate, leading to reduced MGO in L-799. The downregulation of necrosis-related genes with reduced cell death in L-799 shows the potential of priming in maintaining cell viability under stress. Furthermore, upregulated expression of SnRK1.1, SnRK2.6 genes and KIN10 protein levels, with enhanced autophagy markers (ATGs, MDC-stained bodies, lipidated-ATG8), confirmed improved autophagy in melatonin-primed L-799 under stress. Despite lowered ABA, melatonin-mediated MGO homeostasis likely activated MAPK6, inducing autophagy independent of ABA in stressed plants. Conversely, Suraj, with higher endogenous melatonin and inherent stress tolerance, showed limited response to priming. Thus, the study illustrates melatonins role in regulating MGO homeostasis and autophagy under drought stress in cotton.