Label-free quantitative proteomic analysis of Panax ginseng leaves upon exposure to heat stress

Abstract

BackgroundGinseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above 25°C. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level.MethodsWe used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress.ResultsThe results showed a reduction in photosynthetic efficiency on heat treatment (35°C) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated.ConclusionThese results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community.

Spatial protein expression of Panax ginseng by in-depth proteomic analysis for ginsenoside biosynthesis and transportation

“…Ginseng has remarkable and powerful medicinal effects, which can relieve fatigue, regulate blood pressure and endocrine, as well as protect the heart. It can also be used to restore vitality, regulate the body and prolong life for the weak patients [ [1] , [2] , [3] , [4] , [5] ]. Ginseng usually takes 4–6 yrs to mature, and the requirements for the soil, temperature, precipitation, and illumination of the growing environment are high, which further reflected their preciousness and rarity.…”

Optimization of Protein Isolation and Label-Free Quantitative Proteomic Analysis in Four Different Tissues of Korean Ginseng

“…The TCA/acetone method was carried out as described previously [ 7 , 41 ]. Briefly, the OS was incubated with 4 volumes of 12.5% ( w / v ) TCA/acetone containing 0.07% ( v / v ) β -mercaptoethanol for 1 h at −20 °C and then centrifuged at 16,000× g for 10 min at 4 °C to obtain protein pellets.…”

“…In addition, efforts have also been made to improve our understanding of ginseng at the protein level by utilizing proteomics approaches. Studies have focused on identifying stress-responsive and ginsenoside biosynthesis-related proteins, while some studies have concentrated on comparing and analyzing proteins from different ginseng parts and species [ 5 , 6 , 7 , 8 ]. However, a number of these studies used one or two tissues and were based on two-dimensional gel electrophoresis (2-DE) analysis, limiting the comprehensiveness of their proteome data [ 5 , 6 , 9 ].…”

“…Advancements in proteomics approaches have facilitated the proteome analysis of various plants; however, difficulties in extracting relatively pure ginseng proteins have remained a primary obstacle [ 15 ]. Up to now, TCA/acetone method has been extensively used for extracting total ginseng proteins [ 7 ] while TCA precipitation and phenol extraction have been moderately employed to isolate ginseng proteins for 2-DE analysis [ 16 ]. Nonetheless, the efficiency of these methods has been tested on one or two ginseng tissues only, hindering their wide acceptability in ginseng proteome analysis [ 7 , 17 , 18 ].…”

“…Up to now, TCA/acetone method has been extensively used for extracting total ginseng proteins [ 7 ] while TCA precipitation and phenol extraction have been moderately employed to isolate ginseng proteins for 2-DE analysis [ 16 ]. Nonetheless, the efficiency of these methods has been tested on one or two ginseng tissues only, hindering their wide acceptability in ginseng proteome analysis [ 7 , 17 , 18 ]. Therefore, the development of a universal ginseng protein isolation method is a prerequisite for high-throughput ginseng proteome analysis.…”