Proteomic analysis of Potentilla fruticosa L. leaves by iTRAQ reveals responses to heat stress.

Abstract

High temperature is an important environmental factor that affects plant growth and crop yield. Potentilla fruticosa L. has a developed root system and characteristics of resistance to several stresses (e.g., high temperature, cold, drought) that are shared by native shrubs in the north and west of China. To investigate thermotolerance mechanisms in P. fruticosa, 3-year-old plants were subjected to a high temperature of 42°C for 1, 2, and 3 days respectively before analysis. Then, we studied changes in cell ultrastructure using electron microscopy and investigated physiological changes in the leaves of P. fruticosa. Additionally, we used isobaric tags for relative and absolute quantification (iTRAQ) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to study proteomic changes in P. fruticosa leaves after 3 d of 42°C heat stress. we found that the cell membrane and structure of chloroplasts, especially the thylakoids in P. fruticosa leaves, was destroyed by a high temperature stress, which might affect the photosynthesis in this species. We identified 35 up-regulated and 23 down-regulated proteins after the heat treatment. Gene Ontology (GO) analysis indicated that these 58 differentially abundant proteins were involved mainly in protein synthesis, protein folding and degradation, abiotic stress defense, photosynthesis, RNA process, signal transduction, and other functions. The 58 proteins fell into different categories based on their subcellular localization mainly in the chloroplast envelope, cytoplasm, nucleus, cytosol, chloroplast, mitochondrion and cell membrane. Five proteins were selected for analysis at the mRNA level; this analysis showed that gene transcription levels were not completely consistent with protein abundance. These results provide valuable information for Potentilla thermotolerance breeding.