Alleviation of arsenic toxicity in pepper plants by aminolevulinic acid and heme through modulating its sequestration and distribution within cell organelles

Abstract

Aminolevulinic acid (ALA) is essential for chlorophyll and heme synthesis. However, whether heme interacts with ALA to elicit antioxidants in arsenic (As)-exposed plants is still unknown. ALA was applied daily to pepper plants for 3 days prior to beginning As stress (As-S). Then, As-S was initiated for 14 days by employing sodium hydrogen arsenate heptahydrate (0.1 mM AsV). Arsenic treatment decreased photosynthetic pigments (chl a by 38% and chl b by 28%), biomass by 24%, and heme by 47% content, but it elevated contents of malondialdehyde (MDA) by 3.3-fold, hydrogen peroxide (H₂O₂) by 2.3-fold, glutathione (GSH), methylglyoxal (MG), and phytochelatins (PCs) and electrolyte leakage (EL) by 2.3-fold along with enhanced subcellular As concentration in the pepper plant's roots and leaves. The supplementation of ALA to the As-S-pepper seedlings enhanced the amount of chlorophyll, heme content, and antioxidant enzyme activity as well as plant growth, while it reduced the levels of H₂O₂, MDA, and EL. ALA boosted GSH and phytochelates (PCs) in the As-S-seedlings by controlling As sequestration and rendering it harmless. The addition of ALA enhanced the amount of As that accumulated in the root vacuoles and reduced the poisonousness of the soluble As in the vacuoles. The ALA treatment facilitated the deposition and fixation of As in the vacuoles and cell walls, thereby reducing the transport of As to other cell organelles. This mechanism may have contributed to the observed decrease in As accumulation in the leaves. The administration of 0.5 mM hemin (H) (a source of heme) significantly enhanced ALA-induced arsenic stress tolerance. Hemopexin (Hx, 0.4 μg L^-1), a heme scavenger, was treated with the As-S plants along with ALA and ALA + H to observe if heme was a factor in ALA's increased As-S tolerance. Heme synthesis/accumulation in the pepper plants was reduced by Hx, which counteracted the positive effects of ALA. Supplementation of H along with ALA + Hx reversed the negative effects of Hx, demonstrating that heme is required for ALA-induced seedling As-S tolerance.

Keywords: 5-Aminolevulinic acid; Antioxidant system; Arsenic toxicity; Hemin; Pepper.