Abstract
Main conclusion
Two trans-isopentenyl diphosphate synthase and one squalene synthase genes were identified and proved to be involved in the triterpenoid biosynthesis in Platycodon grandiflorus.
Platycodon grandiflorus is a commonly used traditional Chinese medicine. The main bioactive compounds of P. grandiflorus are triterpenoid saponins. The biosynthetic pathway of triterpenoid saponins in P. grandiflorus has been preliminarily explored. However, limited functional information on related genes has been reported. A total of three trans-isopentenyl diphosphate synthases (trans-IDSs) genes (PgFPPS, PgGGPPS1 and PgGGPPS2) and one squalene synthase (SQS) gene (PgSQS) in P. grandiflorus were screened and identified from transcriptome dataset. Subcellular localization of the proteins was defined based on the analysis of GFP-tagged. The activity of genes was verified in Escherichia coli, demonstrating that recombinant PgFPPS catalysed the production of farnesyl diphosphate. PgGGPPS1 produced geranylgeranyl diphosphate, whereas PgGGPPS2 did not exhibit catalytic activity. By structural identification of encoding genes, a transmembrane region was found at the C-terminus of the PgSQS gene, which produced an insoluble protein when expressed in E. coli but showed no apparent effect on the enzyme function. Furthermore, some triterpenoid saponin synthesis-related genes were discovered by combining the component content and the gene expression assays at the five growth stages of P. grandiflorus seedlings. The accumulation of active components in P. grandiflorus was closely associated with the expression level of genes related to the synthesis pathway.
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Data availability
Transcript dataset for gene screening is e available in the NCBI Sequence Read Archive (SRA repository), under the accession number PRJNA688328.
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Funding
This study was supported by the National Natural Science Foundation of China (Grant No. U21A20406) and the Key project at the central government level: The ability establishment of sustainable use for valuable Chinese medicine resources (Grant 2060302-2203-02). Excellent Young Scholars Project of Natural Science Foundation of Anhui Province in China (grant numbers 2208085Y30). Science Research Project at the Universities of Anhui Province for Distinguished Young Scholars (2023AH020036).
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HY designed the experiments, completed partial experiments, and completed the manuscript. BC, JL, and ND provided experimental help. XC, JW, and HP performed partial data and literature searches. LZand SG provided experimental design ideas and modified the manuscript to ensure the precision of the entire experiment. All authors reviewed the final manuscript before submission.
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Yu, H., Chen, B., Li, J. et al. Identification and functional characterization of two trans-isopentenyl diphosphate synthases and one squalene synthase involved in triterpenoid biosynthesis in Platycodon grandiflorus. Planta 258, 115 (2023). https://doi.org/10.1007/s00425-023-04273-2
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DOI: https://doi.org/10.1007/s00425-023-04273-2