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Differential gene expression levels, chemical profiles, and biological activities of Pueraria candollei var. mirifica callus cultures at different growth stages

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Abstract

In vitro callus culture may provide a practical alternative for biomass production and medicinal compounds. The study of gene expression, chemical profile, and bioactivity at different growth stages of callus was conducted to determine the most suitable stage of callus based on the production requirements and provided important data for further study of the biosynthetic pathway. In this study, green (G), yellow (Y), orange (O), and brown (B) callus cultures of Pueraria candollei var. mirifica (PM) were collected at 30, 45, 60, and 75 days after subculture, respectively, and their gene expression levels, chemical profiles, and bioactivities were compared. The complexity and amount of constituents increased with culture time and increasing daidzein and genistein levels until the O callus stage, which was correlated with the constitutive expression of genes involved in the general isoflavonoid biosynthetic pathway at all growth stages. In contrast, the downstream biosynthetic genes encoding cytochrome P450 subfamily 81E and a putative prenyltransferase, which are responsible for diversifying the isoflavones to PM-specific secondary products, were significantly upregulated in the O callus. The observed color changes of the PM callus cultures were attributed to increases in daidzein and genistein levels. The callus cultures exhibited increased antioxidant activity with increasing age, reaching a highest half maximal inhibitory concentration of 0.30 mg/mL in the O callus. The cultures also exhibited cytotoxic activity, with the O and Y callus extracts (each at 3.13 μg/mL) showing significant activity against HepG2 cells (43.3% cell death) and MDA-MD-231 cells (40.1% cell death). These findings demonstrate that age significantly affects the chemical profiles and bioactivities of PM callus cultures.

Key message

Age of Pueraria candollei var. mirifica callus affects its chemical profiles and bioactivities, and is related to isoflavonoids biosynthetic pathway.

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Acknowledgements

This research was supported by the Rachadapisek Sompote Fund for Postdoctoral Fellowships from the Graduate School of Chulalongkorn University, CU’s Natural Product Biotechnology Research Unit, and PMU-C of the Office of the National Higher Education Science Research and Innovation Policy Council. The authors would like to thank Enago (www.enago.com) for the English language review.

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Authors and Affiliations

  1. Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Dolly Rani, Wanchai De-Eknamkul & Sornkanok Vimolmangkang

  2. Graduate Program in Pharmaceutical Science and Technology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Khwanlada Kobtrakul

  3. Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Jittima Amie Luckanagul

  4. Research Unit for Plant-Produced Pharmaceuticals, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Jittima Amie Luckanagul & Sornkanok Vimolmangkang

  5. Department of Food and Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Wuttinont Thaweesest & Pornchai Rojsitthisak

  6. Research Unit for Natural Product Biotechnology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Wanchai De-Eknamkul

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  1. Dolly Rani
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Contributions

DR performed the experiments, analyzed the data, and wrote the manuscript. KK performed the experiments. JAL, WT, and PR executed the cytotoxic experiment. WD helped in the analysis of data and revised the manuscript. SV designed the experiment, analyzed the data, and wrote the manuscript.

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Correspondence to Sornkanok Vimolmangkang.

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Communicated by Sergio Rosales-Mendoza.

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Rani, D., Kobtrakul, K., Luckanagul, J.A. et al. Differential gene expression levels, chemical profiles, and biological activities of Pueraria candollei var. mirifica callus cultures at different growth stages. Plant Cell Tiss Organ Cult 147, 61–72 (2021). https://doi.org/10.1007/s11240-021-02105-3

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