Abstract
Plant tissue culture is controllable and sustainable production system of functional ingredients. Pueraria candollei is used as an herbal medicine to alleviate postmenopausal symptoms. However, conventional sources are limited by prolonged cultivation periods and inconsistent quality. Cell suspension culture of P. candollei was aimed at enhancing and controlling the production of isoflavonoids and deoxymiroestrol. This was achieved by employing 2-hydroxypropyl-β-cyclodextrin (HPBCD) and cellulase as elicitors. The effect of elicitation on estrogenic and anti-inflammatory properties of P. candollei cell extracts was assessed. HPBCD elicitation increases three times the total isoflavonoid accumulation, while cellulase elicitation resulted in 2.5-fold increase in deoxymiroestrol production compared with controls, observed at day 20. Furthermore, HPBCD results in the release of isoflavonoids particularly daidzein (61.30 ± 0.26 µg/mL) into the medium. In both the elicitation groups, the upregulation of genes associated with isoflavonoid and deoxymiroestrol biosynthetic pathways was observed. Specifically, CHS gene transcription was induced within six days. On days 12 and 20, IFS and HID, which convert chalcones to isoflavonoids, were significantly upregulated. Additionally, the upregulation of CYP81E, IFR, and PT-1, which are involved in chromene production, was observed on day 3. HPBCD treatment exhibited a more potent effect on the expression of all the examined genes than cellulase. HPBCD treated extract led to increased estrogenic and anti-inflammatory activities, as evidenced by modulation of gene expression related to these pathways. In summary, HPBCD and elicitation techniques can be strategically employed to improve both quality and productivity of P. candollei cells as a source of functional ingredients.
Key Message
Cellulase increases deoxymiroestrol production in Pueraria candollei cells and 2-hydroxypropyl-β-cyclodextrin has affected cells to release isoflavonoids into the media. Both elicitors enhance the pharmacological activities of P. candollei cells extracts.
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The supporting documentation showing the findings of this study is available from the corresponding author upon reasonable request.
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Acknowledgements
This research project is funded by National Research Council of Thailand (NRCT):
(Contract No. N41A650080) and Faculty of Pharmaceutical Sciences, Khon Kaen University.
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All authors contributed to the study conception and design. WSF carried out the plant, HPLC, ELISA, cells culture experiments and statistical analysis. WSF wrote original draft and visualization. GY and WP reviewed and edited the manuscript. Supervision and critical revision were done by WP. All authors read and approved the final manuscript.
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Sae-Foo, W., Yusakul, G. & Putalun, W. Enhancement of isoflavonoid production and release in Pueraria candollei cell suspension culture using elicitors for improving pharmacological activities. Plant Cell Tiss Organ Cult 156, 99 (2024). https://doi.org/10.1007/s11240-024-02725-5
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DOI: https://doi.org/10.1007/s11240-024-02725-5