Jilin Ginseng Research Articles

Identification of PgRg1-3 Gene for Ginsenoside Rg1 Biosynthesis as Revealed by Combining Genome-Wide Association Study and Gene Co-Expression Network Analysis of Jilin Ginseng Core Collection.

Ginseng, an important medicinal plant, is characterized by its main active component, ginsenosides. Among more than 40 ginsenosides, Rg1 is one of the ginsenosides used for measuring the quality of ginseng. Therefore, the identification and characterization of genes for Rg1 biosynthesis are important to elucidate the molecular basis of Rg1 biosynthesis. In this study, we utilized 39,327 SNPs and the corresponding Rg1 content from 344 core ginseng cultivars from Jilin Province. We conducted a genome-wide association study (GWAS) combining weighted gene co-expression network analysis (WGCNA), SNP-Rg1 content association analysis, and gene co-expression network analysis; three candidate Rg1 genes (PgRg1-1, PgRg1-2, and PgRg1-3) and one crucial candidate gene (PgRg1-3) were identified. Functional validation of PgRg1-3 was performed using methyl jasmonate (MeJA) regulation and RNAi, confirming that this gene regulates Rg1 biosynthesis. The spatial-temporal expression patterns of the PgRg1-3 gene and known key enzyme genes involved in ginsenoside biosynthesis differ. Furthermore, variations in their networks have a significant impact on Rg1 biosynthesis. This study established an accurate and efficient method for identifying candidate genes, cloned a novel gene controlling Rg1 biosynthesis, and identified 73 SNPs significantly associated with Rg1 content. This provides genetic resources and effective tools for further exploring the molecular mechanisms of Rg1 biosynthesis and molecular breeding.

Transcriptome analysis of MYB transcription factors family and PgMYB genes involved in salt stress resistance in Panax ginseng

BackgroundAs the king of all herbs, the medicinal value of ginseng is self-evident. The perennial nature of ginseng causes its quality to be influenced by various factors, one of which is the soil environment. During plant growth and development, MYB transcription factors play an important role in responding to abiotic stresses and regulating the synthesis of secondary metabolites. However, there are relatively few reports on the MYB transcription factor family in Panax ginseng.ResultsThis study identified 420 PgMYB transcripts under 117 genes ID in the Jilin ginseng transcriptome database. Phylogenetic analysis showed that PgMYB transcripts in Jilin ginseng were classified into 19 functional subclasses. The GO annotation result indicated that the functional differentiation of PgMYB transcripts was annotated to 11 functional nodes at GO Level 2 in ginseng. Expression pattern analysis of PgMYB transcripts based on the expression data (TPM) that PgMYB transcripts were revealed spatiotemporally specific in expression patterns. We performed a weighted network co-expression network analysis on the expression of PgMYB transcripts from different samples. The co-expression network containing 51 PgMYB transcripts was formed under a soft threshold of 0.85, revealing the reciprocal relationship of PgMYB in ginseng. Treatment of adventitious roots of ginseng with different concentrations of NaCl revealed four up-regulated expression of PgMYB transcripts that can candidate genes for salt resistance studies in ginseng.ConclusionsThe present findings provide data resources for the subsequent study of the functions of MYB transcription factor family members in ginseng, and provide an experimental basis for the anti-salt functions of MYB transcription factors in Panax ginseng.

Transcriptome-wide characterization, evolutionary analysis, and expression pattern analysis of the NF-Y transcription factor gene family and salt stress response in Panax ginseng

Jilin ginseng (Panax ginseng C. A. Meyer) has a long history of medicinal use worldwide. The quality of ginseng is governed by a variety of internal and external factors. Nuclear factor Y (NF-Y), an important transcription factor in eukaryotes, plays a crucial role in the plant response to abiotic stresses by binding to a specific promoter, the CCAAT box. However, the NF-Y gene family has not been reported in Panax ginseng. In this study, 115 PgNF-Y transcripts with 40 gene IDs were identified from the Jilin ginseng transcriptome database. These genes were classified into the PgNF-YA (13), PgNF-YB (14), and PgNF-YC (13) subgroups according to their subunit types, and their nucleotide sequence lengths, structural domain information, and amino acid sequence lengths were analyzed. The phylogenetic analysis showed that the 79 PgNF-Y transcripts with complete ORFs were divided into three subfamilies, NF-YA, NF-YB, and NF-YC. PgNF-Y was annotated to eight subclasses under three major functions (BP, MF, and CC) by GO annotation, indicating that these transcripts perform different functions in ginseng growth and development. Expression pattern analysis of the roots of 42 farm cultivars, 14 different tissues of 4-year-old ginseng plants, and the roots of 4 different-ages of ginseng plants showed that PgNF-Y gene expression differed across lineages and had spatiotemporal specificity. Coexpression network analysis showed that PgNF-Ys acted synergistically with each other in Jilin ginseng. In addition, the analysis of the response of PgNF-YB09, PgNF-YC02, and PgNF-YC07-04 genes to salt stress treatment was investigated by fluorescence quantitative PCR. The expression of these genes increased after salt stress treatment, indicating that they may be involved in the regulation of the response to salt stresses in ginseng. These results provide important functional genetic resources for the improvement and gene breeding of ginseng in the future.Conclusions: This study fills a knowledge gap regarding the NF-Y gene family in ginseng, provides systematic theoretical support for subsequent research on PgNF-Y genes, and provides data resources for resistance to salt stress in ginseng.

Six weeks of Jilin ginseng root supplementation attenuates drop jump-related muscle injury markers in healthy female college students.

Eccentric muscle contraction is an inherent component of numerous sporting movements but can result in muscle fatigue and injury, especially when engaging in unfamiliar exercise, which requires pharmacological intervention. Jilin ginseng root (GS) has been used to protect muscles and reduce the risk of exercise injury. In this study, we sought to examine and demonstrate the effectiveness of using GS in preventing muscle stiffness and reducing the risk of exercise injury in women. Twenty females were randomly assigned to GS and placebo groups. Body composition, serum biochemistry index, kinematics, and endurance exercise tests were measured at two time point presupplementation and 6 weeks after supplementation. The major compounds of GS were characterized using a high-performance liquid chromatograph with a gradient delivery system (HPLC). After 6 weeks of supplementation, the GS group exhibited significant increases in the serum levels of free fatty acids and glucose as well as greater maximum oxygen consumption (VO2 max, mL min-1 kg-1) compared with the placebo group in an exhaustive biking test. Following drop jump tests, the jump height and reactive strength index were increased in the GS group after completing 70 DJs. In addition, subjects in the GS group also showed decreased knee and ankle stiffness in DJs, leading to reduced fatigue associated with eccentric movement. GS supplementation leads to ameliorates drop jump muscle stiffness and fatigue in females and is to be used as a nutrient supplement to reduce the risk of musculoskeletal system injuries when performing drop jumps.

The bHLH gene family and its response to saline stress in Jilin ginseng, Panax ginseng C.A. Meyer.

Basic helix-loop-helix (bHLH) gene family is a gene family of transcription factors that plays essential roles in plantgrowth and development, secondary metabolism and response to biotic and abiotic stresses. Therefore, a comprehensive knowledge of the bHLH gene family is paramount to understand the molecular mechanisms underlying these processes and develop advanced technologies to manipulate the processes efficiently. Ginseng, Panax ginseng C.A. Meyer, is a well-known medicinal herb; however, little is known about the bHLH genes (PgbHLH) in the species. Here, we identified 137 PgbHLH genes from Jilin ginseng cultivar, Damaya, widely cultivated in Jilin, China, of which 50 are newly identified by pan-genome analysis. These 137 PgbHLH genes were phylogenetically classified into 26 subfamilies, suggesting their sequence diversification. They are alternatively spliced into 366 transcripts in a 4-year-old plant and involved in 11 functional subcategories of the gene ontology, indicating their functional differentiation in ginseng. The expressions of the PgbHLH genes dramatically vary spatio-temporally and across 42 genotypes, but they are still somehow functionally correlated. Moreover, the PgbHLH gene family, at least some of its genes, is shown to have roles in plant response to the abiotic stress of saline. These results provide a new insight into the evolution and functional differentiation of the bHLH gene family in plants, new bHLH genes to the PgbHLH gene family, and saline stress-responsive genes for genetic improvement in ginseng and other plant species.

Transcriptome-Wide Identification, Evolutionary Analysis, and GA Stress Response of the GRAS Gene Family in Panax ginseng C. A. Meyer.

GRAS transcription factors are a kind of plant-specific transcription factor that have been found in a variety of plants. According to previous studies, GRAS proteins are widely involved in the physiological processes of plant signal transduction, stress, growth and development. The Jilin ginseng (Panax ginseng C.A. Meyer) is a heterogeneous tetraploid perennial herb of the Araliaceae family, ginseng genus. Important information regarding the GRAS transcription factors has not been reported in ginseng. In this study, 59 Panax ginseng GRAS (PgGRAS) genes were obtained from the Jilin ginseng transcriptome data and divided into 13 sub-families according to the classification of Arabidopsis thaliana. Through systematic evolution, structural variation, function and gene expression analysis, we further reveal GRAS’s potential function in plant growth processes and its stress response. The expression of PgGRAS genes responding to gibberellin acids (GAs) suggests that these genes could be activated after application concentration of GA. The qPCR analysis result shows that four PgGRAS genes belonging to the DELLA sub-family potentially have important roles in the GA stress response of ginseng hairy roots. This study provides not only a preliminary exploration of the potential functions of the GRAS genes in ginseng, but also valuable data for further exploration of the candidate PgGRAS genes of GA signaling in Jilin ginseng, especially their roles in ginseng hairy root development and GA stress response.

Characterization of the complete chloroplast genome of Jilin ginseng (Panax ginseng C. A. Meyer) using next generation sequencing

Jilin ginseng, Panax ginseng C. A. Meyer, belongs to the Araliaceae family. It is known as the number one medicinal herb and one of the three native treasures in Northern China and has been cultivated in China for over 2000 years. Jilin ginseng is the staple ginseng in China, which contributes to 85% of the ginseng production in the country, thus becoming one of the most important, rapidly increasing industries in both the Province of Jilin and China. In this study, the complete chloroplast genome of Panax ginseng C. A. Meyer was determined by the next generation sequencing. The complete chloroplast genome of Jilin ginseng (P. ginseng C. A. Meyer) was 156,286 bp in length and displays a typical quadripartite structure of the large (LSC, 87,127 bp) and small (SSC, 18,329 bp) single-copy regions, separated by a pair of inverted repeat regions (IRs, 25,415 bp each). It harbors 132 functional genes, including 132 protein-coding genes, 37 transfer RNA, and 8 ribosomal RNA genes species. The overall nucleotide composition was: 30.6% A, 31.3% T, 19.4% C, and 18.7% G, with a total G + C content of 38.1%. Phylogenetic relationship analysis shows that P. ginseng closely related to Panax quinquefolius L.

Evolution, functional differentiation, and co-expression of the RLK gene family revealed in Jilin ginseng, Panax ginseng C.A. Meyer

Most genes in a genome exist in the form of a gene family; therefore, it is necessary to have knowledge of how a gene family functions to comprehensively understand organismal biology. The receptor-like kinase (RLK)-encoding gene family is one of the most important gene families in plants. It plays important roles in biotic and abiotic stress tolerances, and growth and development. However, little is known about the functional differentiation and relationships among the gene members within a gene family in plants. This study has isolated 563 RLK genes (designated as PgRLK genes) expressed in Jilin ginseng (Panax ginseng C.A. Meyer), investigated their evolution, and deciphered their functional diversification and relationships. The PgRLK gene family is highly diverged and formed into eight types. The LRR type is the earliest and most prevalent, while only the Lec type originated after P. ginseng evolved. Furthermore, although the members of the PgRLK gene family all encode receptor-like protein kinases and share conservative domains, they are functionally very diverse, participating in numerous biological processes. The expressions of different members of the PgRLK gene family are extremely variable within a tissue, at a developmental stage and in the same cultivar, but most of the genes tend to express correlatively, forming a co-expression network. These results not only provide a deeper and comprehensive understanding of the evolution, functional differentiation and correlation of a gene family in plants, but also an RLK genic resource useful for enhanced ginseng genetic improvement.

Functional differentiation and spatial-temporal co-expression networks of the NBS-encoding gene family in Jilin ginseng, Panax ginseng C.A. Meyer.

Ginseng, Panax ginseng C.A. Meyer, is one of the most important medicinal plants for human health and medicine. It has been documented that over 80% of genes conferring resistance to bacteria, viruses, fungi and nematodes are contributed by the nucleotide binding site (NBS)-encoding gene family. Therefore, identification and characterization of NBS genes expressed in ginseng are paramount to its genetic improvement and breeding. However, little is known about the NBS-encoding genes in ginseng. Here we report genome-wide identification and systems analysis of the NBS genes actively expressed in ginseng (PgNBS genes). Four hundred twelve PgNBS gene transcripts, derived from 284 gene models, were identified from the transcriptomes of 14 ginseng tissues. These genes were classified into eight types, including TNL, TN, CNL, CN, NL, N, RPW8-NL and RPW8-N. Seven conserved motifs were identified in both the Toll/interleukine-1 receptor (TIR) and coiled-coil (CC) typed genes whereas six were identified in the RPW8 typed genes. Phylogenetic analysis showed that the PgNBS gene family is an ancient family, with a vast majority of its genes originated before ginseng originated. In spite of their belonging to a family, the PgNBS genes have functionally dramatically differentiated and been categorized into numerous functional categories. The expressions of the across tissues, different aged roots and the roots of different genotypes. However, they are coordinating in expression, forming a single co-expression network. These results provide a deeper understanding of the origin, evolution and functional differentiation and expression dynamics of the NBS-encoding gene family in plants in general and in ginseng particularly, and a NBS gene toolkit useful for isolation and characterization of disease resistance genes and for enhanced disease resistance breeding in ginseng and related species.

Characterization of tissue expression of ginsenoside biosynthetic gene expression in Panax ginseng

The study is aimed to characterize the tissue expression of 10 key ginsenoside biosynthetic genes using bioinformatics method and real-time quantitative PCR. Heatmap and cluster analysis of 10 ginsenoside biosynthetic genes were performed in four-year-old Jilin ginseng. Using real-time quantitative PCR, the expression correlation of 10 key genes involved in ginsenoside biosynthesis was analyzed in different organs of four-year-old Jilin ginseng including, tissue culture seedling and adventitious root. Pearson correlation was used to analyze the relation between those 10 key genes involved in ginsenoside biosynthesis. The results showed that β-AS and CYP716A52v2 were expressed highly in root of Jilin ginseng and ginseng culture seedling, which was consistent with Ro distribution. In addition, CYP716A53v2 and CYP716A47 which involved in dammarane type ginsenoside biosynthesis were positively correlated, which revealed that the difference of ginsenoside distribution was caused by transport system.

Correlation of gene expression related to amount of ginseng saponin in 15 tissues and 6 kinds of ginseng saponin biosynthesis

Fifteen tissues of 4-year-old fruit repining stage Jilin ginseng were chosen as materials, six kinds of monomer saponins (ginsenosides Rg1, Re, Rb1, Rc, Rb2 and Rd) content in 15 tissues was measured by HPLC and vanillin-sulfuric acid method. The relative expression of FPS, SQS, SQE, OSC, β-AS and P450 genes in 15 tissues was analyzed by real-time PCR. The correlations between ginseng saponin content in 15 tissues of Jilin ginseng and biosynthetic pathway -related genes were obtained. The results showed that was a synergistic increase and decrease trend of positive linear correlation among six kinds of monomer saponin content, and there was a significantly (P < 0.01) positive correlation between monomer saponin content and total saponins content. Monomer saponin content and 6 kinds of enzyme gene correlation were different. Biosynthesis of ginseng total saponins and monomer saponin were regulated by six kinds of participation ginsenoside biosynthesis enzyme genes, the expression of these six kinds of genes in different tissues of ginseng showed collaborative increase and decrease trend, and regulated biosynthesis of ginseng ginsenoside by group coordinative manner.

A plant-transformation-competent BIBAC library of ginseng (Panax ginseng C. A. Meyer) for functional genomics research and characterization of genes involved in ginsenoside biosynthesis

Ginseng (Panax ginseng C. A. Mey.) is widely used as a major medicinal herb and as a feedstock for the medicine, beverage, food, cosmetic, etc. industries, in China and several other Asian countries. However, limited research has been accomplished into its genetics, genomics and breeding. To clone, characterize and utilize the genes of economic importance in the species, we have developed a large-insert plant-transformation-competent binary bacterial artificial chromosome (BIBAC) library for Jilin ginseng cv. Damaya. The library contains 141,312 clones, with an average insert size of 110 kb, each likely containing approximately 20–30 genes. The clones of the library have all been arrayed in 384-well microplates and permanently archived. We screened the library and identified BIBAC clones containing nine genes likely involved in the biosynthesis pathway of ginsenosides—the major medicinally effective compounds of ginseng—with approximately four BIBACs per gene. This result further verified the quality of the library and demonstrated its utility in cloning, characterization and utilization of economically important genes in ginseng. Furthermore, since the library is cloned in a plant-transformation-competent BIBAC vector (pCLD04541) that can be directly transformed in a variety of plants via both the Agrobacterium-mediated method and the particle bombardment method, we have also demonstrated the stability of large-insert ginseng DNA BIBACs in different Agrobacterium strains, which is crucial to large-insert BIBAC transformation in plants. Therefore, the Jilin ginseng BIBAC library provides resources and tools useful for functional genomics research, and cloning, characterization and utilization of economically important genes in the species.

Simultaneous Determination of Trace Antimony and Bismuth in Jilin Ginseng with Microwave Digestion Hydride Generation Atomic Fluorescence Spectrometry

A robust, accurate and sensitive analytical procedure was developed for the simultaneous determination of trace Antimony and Bismuth in Jilin ginseng by hydride generation atomic fluorescence spectrometry. The parameters were studied systematically, such as acid concentration of the reaction medium, flow rate of the carrier gas and shield gas, the atomizer of height, etc. Ascorbic acid and thiourea were used as reducer or masking agents to enhance the generation efficiency of the volatile species of Sb and Bi. In the presence of thiourea and ascorbic acid, the influences of some coexisting elements on the determination of antimony and bismuth were investigated.

Separation and determination of chemical constituents in the volatile oil of three traditional Chinese crude drugs

The parts of Jilin ginseng growing on the ground, Radix aucklandiae and the citrus tangerina peels were extracted with ether, and yellow volatile oil 0.31 wt.%, 2.1 wt.% and 1.6 wt.% were obtained respectively. Their chemical constituents were separated and determined by gas chromatography and interfaced with mass spectrometry in an electron impact mode.

A Comparison of Mineral Element Contents Among Different Types of Ginseng Using Inductively Coupled Plasma-Atomic Emission Spectroscopy

Contents of 24 mineral elements in different types of Jilin ginseng were determined for the first time by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The results indicate that the contents vary with ginseng type, location, and place of origin. The total amounts of the essential elements Ca, P, Mg, Fe, Sr, Mn, Zn, Cr, Cu, Ni, V, Co, and Mo in the types of ginseng tested are in the following order: Changbe > Zhujielu > Damaya > Ermaya > Yuanlu. This agrees with the total contents of elements detected. The amount of harmful microelements such as Pb and Cd in all types of ginseng is far below the toxic dose. In this study, the relative error is 1.76-15.38%, the precision is 0.3-11.0%, the detection power is 0.00 X- X × 10 3 μg/g, and the standard recovery is 90-106%. Thus a scientific foundation was provided for the optimum selection of superior types of ginseng.

Quantitative analysis of ginsenosides in fresh Panax ginseng.

TLC, DCC and HPLC were used to study the ginsenoside composition of the main root, lateral root, rhizome, leaves and seeds of Panax ginseng cultivated in Jilin, China. Each of these methods has advantages of its own and the ensemble reveal the special features of Jilin ginseng. Total saponin content of various plant parts in Jilin ginseng showed a mid-range value as compared to those in ginsengs reported in literature. Fresh as well as sun-dried specimens from the same batch possessed a high percentage of Rg1 in the main root and this might account for the traditional preference of this plant part despite its lowest percentage of saponin in the whole plant. Large amounts of polar saponins were also observed in roots and rhizome of fresh Jilin ginseng, the nature and significance of which remained to be investigated.