Ascorbate peroxidase (APX) is a heme-containing enzyme involved in hydrogen peroxide (H2O2) detoxification within the ascorbate–glutathione (AsA–GSH) cycle. In this study, the full-length genomic DNA and cDNA of an APX1 gene (VsAPX1) were cloned and characterized from Vicia sativa. The genomic sequence of VsAPX1 is 2425 bp in length and comprises 10 exons separated by nine introns, with the first intron located within the 5′ untranslated region (5′UTR). The corresponding cDNA is 1010 bp long and includes a 61 bp 5′UTR, a 753 bp open reading frame, and a 196 bp 3′UTR. VsAPX1 encodes a predicted cytosolic APX protein of 250 amino acids, with a molecular weight of 27.1 kDa and a theoretical isoelectric point (pI) of 5.60. Bioinformatics analysis revealed that the deduced VsAPX1 protein shares high sequence similarity with cytosolic APX1 proteins from other plant species, contains conserved APX domains, and clusters within the cytosolic APX clade in phylogenetic analysis. Quantitative real-time PCR analysis showed that VsAPX1 expression exhibits transient and moderate changes in response to abiotic stress and phytohormone treatments. Transcript levels increased at early time points following heat stress (42 °C), abscisic acid, and salicylic acid treatments, and after 4 h of jasmonic acid exposure, whereas hydrogen peroxide treatment resulted in a gradual down-regulation of expression. Overall, this study provides the first molecular and expression characterization of a cytosolic APX1 gene from Vicia sativa and establishes a foundation for future functional analyses of antioxidant genes in this species.
Siam et al. (Sat,) studied this question.