Analysis of differentially expressed proteins and related metabolic pathways in response to lead stress in the leaves of Pogonatherum crinitum

Abstract

Proteomics provides an essential means of explaining the mechanisms underlying gene expression regulation. The proteomic mechanisms by which heavy metal hyperaccumulators respond to lead (Pb) stress remain largely unclear. To this end, we examined Pogonatherum crinitum (Thunb.) Kunth and employed proteomic sequencing technology to screen for differential proteins that respond to Pb stress. The connection between Pb-tolerant proteins in metabolic pathways and their functions were analyzed. Differences in the downstream molecules of Pb-resistant proteins in P. crinitum were also assessed. Furthermore, we utilized Parallel Reaction Monitoring (PRM) technology to validate the selected Pb-tolerant differential proteins across various stress concentration gradients. A total of 5275 protein families were identified, and 118 DEPs were observed between the stressed and control groups, including 76 upregulated and 42 downregulated proteins. Functional annotation analysis using Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes revealed that Pb stress led to the upregulation of 16 proteins within P. crinitum leaves. These proteins were primarily involved in the metabolic processes of energy and carbohydrate metabolism (PcCht1, PcSPS5, PcGME-1, and PcPEP4) as well as protein translation and oxidative stress (PcHSP26.7, PcHSP18, PcCAT3, and PcCAT1). Bioinformatic analysis indicated that DEPs responding to Pb stress were primarily related to the MAPK signaling pathway, amino sugar and nucleotide sugar metabolism, and starch and sucrose metabolism. Pathway analysis revealed maltose, acetylcholine, N-acetylglucosamine, and oxalic acid as the downstream products. Moreover, the levels of these indicators all increased with increasing Pb concentrations. PRM of the 16 DEPs revealed that nine proteins were upregulated under different Pb concentrations. PRM and data-independent acquisition results for the upregulation of these nine DEPs were identical, suggesting the reliability of our analytical outcomes. In conclusion, the upregulation of specific proteins in P. crinitum enables the regulation of glucose metabolism and antioxidant balance within the plant and represents a mechanism underlying its Pb stress response.

Keywords: Lead stress; Metabolic pathways; Parallel reaction monitoring; Pogonatherum crinitum (Thunb.) Kunth; Proteomics.