Functional Characterization of Glutathione Peroxidase Genes Reveals Their Contribution to the Rapid Range Expansion of Amaranthus Palmeri Under Stress Conditions

Amaranthus palmeri is an aggressive, highly invasive weed that thrives across a wide range of adverse environments worldwide; nevertheless, the mechanisms underlying its rapid expansion remain largely unstudied. Glutathione peroxidase (GPX) is a crucial enzyme within the antioxidant defense system, belonging to the phylogenetic conserved family of oxidoreductases present in all living organisms. Despite its significance, the role of GPX in A. palmeri has not been reported. This study identified eight GPX genes (ApGPXs) in A. palmeri through comprehensive bioinformatics and gene expression analyses. The research examined the characteristics, evolutionary relationships, chromosomal mapping, gene structure, subcellular localization, conserved motifs, and cis-acting elements of these genes, as well as their evolutionary conserved functions in relation to Arabidopsis thaliana GPXs and RT-qPCR analysis under various stress conditions. The ApGPXs were distributed across scaffolds (2, 4, and 12) of the A. palmeri genome. Phylogenetic analysis grouped GPX genes into four subgroups, and conserved motifs were found within certain phylogenetic subgroups. We identified Actin8 as the most stable internal reference gene for A. palmeri under diverse stress conditions. Gene expression analysis revealed that ApGPXs participate in both early and late regulatory responses to oxidative stress induced by NaCl, high temperature, osmotic pressure, and glufosinate ammonium. The Arabidopsis GPX mutant (AT4G31870) exhibited a stronger flg22-induced oxidative burst than the wild type, and qPCR confirmed that AtGPXs contribute significantly to glufosinate ammonium stress responses. Evolutionary analysis found high sequence similarity between ApGPX4 and AT1G63460, as well as ApGPX3 and AT4G11600. Also, ApGPX3 and AT4G11600 shared similar expression patterns under glufosinate ammonium stress. This research presents the first gene family study in A. palmeri and provides foundational insights for future studies in this economically critical species. Our findings establish a framework for mitigating A. palmeri’s impact on crop production and exploring ApGPXs in developing herbicide- and stress-tolerant cultivars.