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A Candidate Gene Identified in Converting Platycoside E to Platycodin D by Correlating Main Metabolites Content with Gene Expression Variation between Callus and Leaf of Platycodon grandiflorus

ABSTRACT: Platycodon grandiflorus is a perennial traditional Chinese herb, which has more than 100 compounds including triterpenoid saponins, polysaccharides and phenolic acid etc. In this study, a (L9(34)) orthogonal experiment determined the optimal explants is stem with nodes and the optimum full medium is MS + NAA 1.0 mg/L + 6-BA 0.5 mg/L as best protocol for callus induction of P. grandiflorus.The content of platycodin D, platycoside E and total polysaccharides between callus and plant organs is significantly difference, and the distribution of platycodin D, platycoside E and total polysaccharide in whole plant has been clarified. Meanwhile, the content of platycodin D and total polysaccharide were found higher in calli than in leaves or the whole plant. While, the content of platycoside E in calli is very lower than in leaves. In order to screen candidate genes involved in platycoside E and platycodin D conversion, a RNA-Seq analysis between calli and leaves was performed. Correlating the content of platycodin D and platycoside E between calli and leaves with the gene expression level involving in triterpenoid saponin biosynthesis pathway, four cluster genes with high amino acid similarity were screened as the putative sequences of β-Glucosidase gene for converting platycoside E to platycodin D. It was found that the expression level of genes encoding enzymes involved in saponins and polysaccharides biosynthesis was co-expression with the content of the corresponding metabolite. Besides, some transcription factors predicted to regulate the expression of key enzymes involving in triterpernoid saponins and polysaccharides biosynthesis pathway of P. grandiflorus.

ORGANISM(S): Platycodon grandiflorus

PROVIDER: GSE153777 | GEO | 2020/07/15

REPOSITORIES: GEO

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Project description:Background: Dendrobium officinale, an endangered Chinese herb, has extensive therapeutic effects and contains bioactive ingredients including a large number of polysaccharides and alkaloids, and minimal flavonoids. Firstly, this study attempts to obtain the protocorm-like bodies of this plant through tissue culture to produce the main secondary metabolites whose distribution in each organelle and protocorm like bodies is analyzed. Then, analysis of the correlation between comparative transcriptome sequence and the metabolite content in different organs enables the discovery of putative genes encoding enzymes involved in the biosynthesis of polysaccharides and alkaloids, and flavonoids. Results: The optimum condition for protocorm-like bodies (PLBs) induction and propagation of D. officinale is established. For protocorm induction, we use the seed as the explant, and the optimum medium formula for PLBs propagation is 1/2 MS + α-NAA 0.5 mg·L-1 +6-BA 1.0 mg·L-1 + 2, 4-D 1.5-2.0 mg·L-1 + potato juice 100 g·L-1. The distribution of polysaccharides, alkaloids and flavonoids in D. officinale organs was clarified. Stems, PLBs and leaves have the highest content of polysaccharides, alkaloids and flavonoids, respectively. PLBs replace organs to produce alkaloids in D. officinale, and naringenin was only produced in stem. Hot water extraction (HWE) method was found outperforming the ultrasound-assisted extraction (UAE) method for polysaccharides from D. officinale. A comparative transcriptome analysis of the protocorm-like bodies and leaves of D. officinale showed genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids biosynthetic pathway were differentially expressed. Putative genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids synthetic pathway were identified. Notably, genes encoding enzymes of strictosidine beta-glucosidase, geissoschizine synthase and vinorine synthase in alkaloids biosynthesis of D. officinale are first reported. Conclusions: Our works, especially the identification of candidate genes encoding enzymes involved in metabolites biosynthesis will help to explore and protect the endangered genetic resources and will also facilitate further analysis of the molecular mechanism of secondary metabolites’ biosynthesis in D. officinale.

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A Candidate Gene Identified in Converting Platycoside E to Platycodin D by Correlating Main Metabolites Content with Gene Expression Variation between Callus and Leaf of Platycodon grandiflorus

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