Melatonin improves the lead tolerance in Plantago ovata by modulating ROS homeostasis, phytohormone status and expression of stress-responsive genes.

key messageMelatonin increases Pb tolerance in P. ovata seedlings via the regulation of growth and stress-related phytohormones, ROS scavenging and genes responsible for melatonin synthesis, metal chelation, and stress defense. Lead (Pb) is a highly toxic heavy metal that accumulates in plants through soil and air contamination and impairs its plant growth and development. Because of its pharmaceutical importance, improvements in Plantago ovata yield against abiotic stresses are necessary. Melatonin (MEL) is a stress-alleviating biostimulator and our results showed a reduction in Pb induced phytotoxicity by enhancing plant growth attributes and balancing protective osmolytes. Pb-induced reactive oxygen species accumulation, including superoxide and peroxide free radicals and their mitigation through enzymatic antioxidants, was demonstrated in presence of MEL. Cell viability and Pb bioaccumulation were determined to understand the extent of cellular damage. Moreover, MEL increased secondary metabolite (flavonoids and anthocyanins) contents by 2-3-fold at the lowest Pb concentrations. Similar increases in the relative expression of genes (PoPAL and PoPPO), which are responsible for the production of non-enzymatic antioxidants, were observed. Notably, the upregulation of the PoCOMT gene up to 4-fold indicates increased melatonin production, as manifested in the phytomelatonin level. MEL supplementation also increased the auxin (IAA) level by 3-fold in the 100 µM Pb treatment group, while the abscisic acid (ABA) level decreased (1.4-fold) and the expression of PoMYB (a stress-related transcription factor) increased (up to 2.66-fold). Additionally, we found extreme downregulation (up to 18-fold) in the relative expression of PoMT 2 (a metal binding thiol compound) with melatonin treatment, which is otherwise upregulated (by 6-fold) during Pb stress. In the current study, these effects collectively revealed that MEL contribute to enhanced plant growth and Pb stress tolerance.