Expression analysis of transcription factor ERF gene family of Panax ginseng / 中国中药杂志

Ethylene responsive factor(ERF), one of the largest families of transcriptional factors in plants, plays a key role in se-condary metabolism of herbal plants. To analyze the expression of ERF family genes, the heat map clustering method was used by analyzing the ginseng transcriptomes of different parts and different growth years. The contents of ginsenosides Rg_1, Re and Rb_1 in various concentrations of MeJA-treated ginseng adventitious roots were determined by UPLC-MS/MS method. The expression of key genes of ginsenoside biosynthesis(DDS, CYP716A47, CYP716A53v2) and ERF family genes in MeJA-treated ginseng adventitious roots were determined by using real-time quantitative PCR. Pearson correlation was adopted to analyze the gene expression pattern of DDS, CYP716A47, CYP716A53v2 gene and ERF family. The results showed that the content of ginseng diol ginsenoside Rb_1 in ginseng adventitious roots treated with different concentrations of MeJA increased, and the content of ginseng triol ginsenoside Rg_1 and Re decreased. It is consistent with the increase of DDS and CYP716A47 expression and the decrease of CYP716A53v2 gene expression. The expression of ERF003, ERF118 and ERF012 genes was significantly positively correlated with CYP716A53v2, but negatively correlated with DDS. While the expression of ERF1B was significantly negatively correlated with CYP716A47.It is proved that ERF003, ERF118 and ERF012 were likely to inhibit the expression of DDS and promote the expression of CYP716A53v2, and ERF1B was likely to inhibit CYP716A47. This work could provide theoretical basis of ERF functional verification of regulating the biosynthesis of ginsenosides.

Expression of saponin biosynthesis related genes in different tissues of / 中国中药杂志

The relationship between saponin content of in different parts of the organization and expression of ginsenoside biosynthesis related gene was obtained by the correlation analysis between saponin content and gene expression. The 14 tissue parts of were studied, six saponins in Samples (ginsenoside Rg₁, Re, Rb₁, Rc, Rb₂ and Rd), group saponins and total saponins were determined by high performance liquid chromatography and vanillin-sulfuric acid colorimetric method. Simultaneously, the expression levels of 7 ginsenoside biosynthesis related genes ( and ) in different tissues of were determined by Real-time fluorescence quantitative PCR. Although 7 kinds of ginsenoside biosynthesis related enzyme gene in the involved in ginsenoside synthesis, the expression of and P450 genes had no significant effect on the content of monosodium saponins, grouping saponins and total saponins, and had significant or extremely significant on the contents of single saponins Re, Rg1, Rb1, Rd, group saponin PPD and PPT, total saponin TMS and total saponin TS (<0.05 or <0.01). The biosynthesis of partial saponins, grouping saponins and total saponins in was affected by the interaction of multiple enzyme genes in the saponin synthesis pathway, the content of saponins in different tissues of was determined by the differences in the expression of key enzymes in the biosynthetic pathway. Therefore, this study further clarified that and was the key enzyme to control the synthesis of saponins in by correlation analysis, the biosynthesis of ginsenosides in was regulated by these five kind of enzymes in cluster co-expression of interaction mode.

Research of genes expression related to ginsenosides biosynthesis by methyl jasmonic acid in ginseng hair roots / 中国中药杂志

In order to obtain the expression of ginsenoside biosynthetic pathway related enzyme gene in ginseng hairy root under the control of elicitors, methyl jasmonate (MeJA) was added exogenously as elicitors. Ginseng hairy root clones induced by 4-year-old ginseng root was used as material, total saponin content in ginseng hairy root before and after MeJA treatment was determined by vanillin-sulfuric acid colorimetry, Meanwhile, relative expression of squalene synthase genes, squalene epoxidase genes, oxidized squalene cyclase genes, dammarenediol synthase genes, β-amyrin synthase genes, cycloartenol synthase genes before and after MeJA treatment were determined by Real-time PCR. The optimum conditions of MeJA which added to ginseng hairy root were obtained, the optimum additional concentration was 6×10⁻⁴ μmol•L⁻¹, the optimum additional time was 22 d, and the optimum action time was 5 d. The addition of MeJA could improve the enzymatic activity of peroxidase (PPD), catalase (CAT) and peroxidase (PPD) in ginseng hairy root. The expression of SQS，SQE，OSC，DS and β-AS genes of ginsenoside biosynthetic pathway increased significantly after MeJA treatment, while the change of CAS gene expression were not significant. The expression of key enzyme SQS，SQE，OSC，DS and β-AS genes in ginsenoside biosynthetic pathway was consistent with the changes of PPD，CAT，PPO enzymatic activity.

Advances in Transcriptomic Studies and Ginsenoside Biosynthesis of American Ginseng / 中草药·英文版

American ginseng (. Panax quinquefolius L.), belonging to the Araliaceae family, is one of the most widely used traditional herbs in the world. Its major bioactive constituents are triterpene saponins known as ginsenosides. Up to date, it is still a big challenge to sequence and assemble the large and repeat-enriched genome of tetraploid American ginseng, using whole genome shotgun (WGS) sequencing strategy. The lack of American ginseng genome information has significantly impeded its genetic and functional genomic studies. With the development of next-generation sequencing (NGS) technologies, sequencing and analysis of transcriptomes have become powerful tools for the discovery of novel genes and elucidation of specific biosynthetic pathways of secondary metabolites. Here we summarized the recent advances in the transcriptomic studies of American ginseng, including high-throughput transcriptome sequencing, assembly, and functional gene annotation and classification. Based on the results of transcriptomic data mining and co-expression analyses, many candidate genes possibly involved in the biosynthetic pathway of ginsenosides have been found, thereby providing an unparalleled opportunity to fully understand the mechanism of ginsenoside biosynthesis and its regulations in American ginseng. Advances in transcriptomic studies will contribute to the molecular breeding and planting management of American ginseng and to the development of novel ginsenoside-type drugs.