Proteomic analysis of Korean ginseng (Panax ginseng C.A. Meyer)

doi:10.1016/j.jchromb.2004.10.063

Journal of Chromatography B

Volume 815, Issues 1–2, 5 February 2005, Pages 147-155

https://doi.org/10.1016/j.jchromb.2004.10.063 Get rights and content

Abstract

Although many reports have been published regarding the pharmacological effects of ginseng, little is known about the biochemical pathways operant in ginsenoside biosynthesis, or the genes involved therein. Proteomics analysis is an approach to elucidate the physiological characteristics and biosynthetic pathways of ginsenosides, main components of ginseng. In this review, we introduced the recent progress in proteomics studies of ginseng (Panax ginseng C.A. Meyer). We briefly reference the genomic analyses of P. ginseng, without which proteomics approaches would have been impossible. Functional genomics studies regarding secondary metabolism in P. ginseng are also introduced here, in order to introduce possible future prospects for further study.

Introduction

Ginseng is a perennial herb of the family Araliaceae. Seven major species of ginseng are distributed throughout East Asia, Central Asia, and North America [1], [2]. Most studies of ginseng have been carried out in Panax ginseng (Asian ginseng), Panax quinquefolius (American ginseng), or Panax japonicus (Japanese ginseng). The roots of P. ginseng have been commonly used in the formulation of tonics in Eastern Asia for over 2000 years. It is believed that the root of P. ginseng is a panacea, i.e., it is both a universal cure and it also promotes longevity. Various clinical and pharmacological effects associated with its use have been reported, such as anti-cancer activity, anti-circulatory shock effects, promotion of hematopoiesis, and modulation of immune functions and cellular metabolic processes on carbohydrates, fats and proteins [3], [4], [5].

It is believed that the pharmacological effects of P. ginseng vary according to the age and species of P. ginseng [6]. For example, it was reported that aged P. ginseng had a more pronounced anti-carcinogenic effect on lung tumors in mice than the unaged plant [7]. It is also believed that different parts of P. ginseng exhibit different properties and medical values [8].

Because P. ginseng must grow for at least 3 years before it begins to exert medically valuable effects, the productivity of P. ginseng can be significantly affected by various environmental factors such as temperature, condition of soil, light intensity, content of water and disease. Therefore, extensive studies have been carried out to elucidate the physiological responses of P. ginseng to environmental changes [9], [10], [11], [12]. Methods for the differentiation and authentication of P. ginseng species and products have been developed in response to commercial demand. Those methods include product analysis using HPLC/mass spectrometry (MS) [6], PCR-restriction fragment length polymorphism [13], [14], [15], [16], [17], and proteome analysis [8].

Ginsenosides are considered to be the main active pharmacological compounds in P. ginseng. Thirty-one ginsenosides have, so far, been isolated from natural and processed P. ginseng roots, and novel ginsenosides continue to be reported [18], [19]. The distribution of ginsenosides varies from species to species [2]. Each ginsenoside has different pharmacological effects, and even one single ginsenoside has been demonstrated to produce multiple effects in the same tissue [1], [20], [21].

Although many reports have been published regarding the pharmacological effects of ginsenosides, little is known about the biochemical pathways operant in ginsenoside biosynthesis, or the genes involved therein. In the effort to delineate these biosynthesis pathways and their regulation, a variety of analytical techniques have been employed, including P. ginseng EST sequence analysis [18].

Though there are many technical challenges and pitfalls, proteomics is becoming an essential technology in the field of biological science. Recent technical improvements in two-dimensional polyacrylamide gel electrophoresis (2-DE) and MS have made it possible to rapidly identify hundreds of proteins and investigate levels of protein expression, subcellular localization, and post-translational modification. Proteomics is applicable to the characterization of individual cells, estimation of genetic variability, phylogeny, characterization of mutants, screening of useful proteins, and the determination of the effects of different factors (e.g., light, heat, cold, and hormones) on plant growth [22], [23].

In this review, we will discuss the proteomics challenges regarding P. ginseng, and recent proteomics studies of P. ginseng. For this purpose, we shall also briefly refer to the genomic analyses of P. ginseng, without which proteomics approaches would have been impossible. Functional genomics studies regarding secondary metabolism in P. ginseng are also introduced here in order to introduce possible future prospects for further study.

Section snippets

Genomics of P. ginseng

Genomic studies of P. ginseng were initiated by the Plant Diversity Research Center (PDRC), which was established in 2000 by the Ministry of Science and Technology in Korea. The PDRC aims at the promotion of systematic management and utilization of plant resources indigenous to the Korean peninsula. P. ginseng has been selected as one of Korea's most important plant resources due to the richness and utility of its secondary metabolites like ginsenosides. However, the amount of genetic

Sample preparation for 2DE

Recent technical improvements in proteomic analysis have made it possible to identify hundreds of proteins, and thus provide information regarding protein expression and cellular regulation. But the application of proteomic analysis to plant samples still has some limitations. Acquiring a smart and reproducible 2-DE gel image remains a difficult proposition, largely due to the interference of compounds such as lignins, polyphenols, tannins, alkaloids and pigments [25]. Therefore, the removal of

Functional genomics for secondary metabolism analysis in P. ginseng

Functional genomic studies of P. ginseng have also been carried out in order to elucidate the genes involved with the secondary metabolism of saponin biosynthesis in P. ginseng. In order to select the hairy root lines of P. ginseng that exhibit better production of specific ginsenosides, DNA microarray and metabolic profiling have been undertaken. In parallel, several laboratories have studied the generation of P. ginseng transgenic plants through genetic manipulation.

Conclusions

Cultivation of ginseng is difficult, and requires a growth period of 4–6 years before roots can be harvested for the use as medicine. This plant should be cultivated under shadowy conditions, and consecutive cultivations in the same soil has to be avoided. Root rot disease caused by fungi, and red skin disease of unknown etiology are serious constraints in the cultivation of P. ginseng.

The root of P. ginseng has been used for thousands of years as a folk medicine in Asian countries. The major

Acknowledgements

This work was supported by a grant (PF003101-03) to YMP and a grant (PF003101-02) to DCY and YPL from the Plant Diversity Research Center of the 21st Century Frontier Research Program, funded by the Ministry of Science and Technology of the Korean government. JRL acknowledges Kyung Hee University Plant Metabolism Research Center funded by the Korea Science and Engineering Foundation and the Genetic Resources and Information Network Center funded by the Ministry of Science and Technology of

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The genus Panax belongs to the taxonomic family Araliaceae and consists of many species that are commonly referred to as ginseng. The plants are perennial herbs that grow mostly in the wild and only a few are cultivated. Geographically, they are mainly distributed in North America and northeast Asia. The species contain bioactive chemicals including ginsenosides that have some clinical evidence for the treatment of medical ailments ranging from the common cold, diabetes, and cancer to neurological disorders. Ginseng is mainly propagated by seeds that can be difficult to germinate and often produce plants that are of variable characteristics and quality. In the natural habitat, the Panax species have a production cycle of 3–7 years from seedling to harvest. Research into the in vitro culture of ginseng cells, tissues, and organs began in the 1960s, and by 1980 methods for large-scale propagation were established. With in vitro culture methodologies, plant production can be controlled in terms of both quality and quantity. In vitro methods of micropropagation can reduce the lengthy plant production cycle and can preserve unique genotypes and phenotypes. Micropropagation also allows a more rapid multiplication of plants of superior quality and genetic uniformity than seedling production. This chapter provides a general overview of ginseng growth characteristics, development, and an update on the major scientific research contributions of the in vitro culture techniques for improving the cultivation of Panax species.
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ReviewProteomic analysis of Korean ginseng (Panax ginseng C.A. Meyer)

Abstract

Introduction

Section snippets

Genomics of P. ginseng

Sample preparation for 2DE

Functional genomics for secondary metabolism analysis in P. ginseng

Conclusions

Acknowledgements

Biochem. Pharmacol.

Phytochemistry

Curr. Opin. Plant Biol.

Methods

Biotech. Adv.

Curr. Opin. Biotech.

Phytochemistry

Plant Physiol. Biochem.

J. Plant Physiol.

Comp. Biochem. Phys. B

Trends Biotechnol.

Plant Cell Rep.

Biochem. Pharmacol.

Int. J. Biochem. Cell Biol.

Jpn. J. Pharmacol.

The Pharmacology of Chinese Herbs

Cancer Causes Control

J. Ethnopharmacol.

Ann. Acad. Med. Singapore

Anal. Chem.

J. Korean Med. Sci.

Proteomics

Planta

Zhong Yao Cai

Zhong Yao Tong Bao

J. Ginseng Res.

Acta Pharmacol. Sin.

Biol. Pharm. Bull.

J. Ginseng Res.

Acta Hortic.

Plant Cell Rep.

Chem. Pharm. Bull. (Tokyo)

Planta Med.

Biochem. Biophys. Res. Commun.

Electrophoresis

Proteomics

Mol. Gen. Genomics

Methods Enzymol.

Review
Proteomic analysis of Korean ginseng (Panax ginseng C.A. Meyer)