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Alteration in the callogenesis, tropane alkaloid formation, and gene expression in Hyoscyamus niger under clinorotation

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Abstract

This study aimed to investigate the effects of clinorotation induced by 2-D clinostat on the growth, tropane alkaloid production, gene expression, antioxidant capacity, and cellular defense responses in the callus tissue of Hyoscyamus niger. Callus induction was conducted by putting hypocotyl explants in the MS culture medium supplemented with 1 mgL−1 2,4-D and 1 mgL−1 BAP growth regulators. The sub-cultured calli were placed on a clinostat for 0, 3, 7, and 10 days (2.24 × 10–5 g on the edge of the callus ring). Clinorotation significantly increased callus fresh weight, dry weight, protein, carbohydrate, and proline contents compared to the control, and their maximum contents were obtained after 7 and 10 days. H2O2 level enhanced under clinorotation with a 76.3% rise after 10 days compared to control and positively affected the atropine (77.1%) and scopolamine (69.2%) productions. Hyoscyamine 6-beta hydroxylase and putrescine N-methyltransferase gene expression involved in the tropane alkaloid biosynthesis were upregulated markedly with 14.2 and 17.1-folds increase after 10 days of clinorotation, respectively. The expressions of jasmonic acid, mitogen-activated protein kinase, and ethylene-responsive element-binding transcription factor were upregulated, and the activity of peroxidase and catalase showed a 72.7 and 80% rise after 10 days. These findings suggest that microgravity can enhance callogenesis by stimulating the ROS level, which can impact the antioxidant enzymes, tropane alkaloid formation, and gene expression.

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Data Availability

The data that support the findings of this study are available from the corresponding author, upon reasonable request.

Abbreviations

JA:

Jasmonic acid

H6H :

Hyoscyamine 6-beta hydroxylase

PMT :

Putrescine N-methyltransferase

MAPK :

Mitogen-activated protein kinase

EREB :

Ethylene-responsive element binding

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Authors and Affiliations

  1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Sara Pourhabibian, Alireza Iranbakhsh & Azadeh Hekmat

  2. Department of Biology, Damghan Branch, Islamic Azad University, Tehran, Iran

    Mostafa Ebadi

  3. Aerospace Research Institute, Ministry of Science Research and Technology, Tehran, 14665-834, Iran

    Halimeh Hassanpour

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  1. Sara Pourhabibian
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Correspondence to Alireza Iranbakhsh.

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Pourhabibian, S., Iranbakhsh, A., Ebadi, M. et al. Alteration in the callogenesis, tropane alkaloid formation, and gene expression in Hyoscyamus niger under clinorotation. Protoplasma 261, 293–302 (2024). https://doi.org/10.1007/s00709-023-01894-y

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