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In vitro micropropagation and conservation of endangered medicinal plant Nepeta asterotricha Rech.f. (Lamiaceae): genetic fidelity, phytochemical and biological assessment

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Abstract

An efficient in vitro protocol was introduced for the conservation of Nepeta asterotricha, a vulnerable and endangered medicinal species found in the central of Iran for the first time. Growth, phytochemical, and biological traits of in vitro regenerated plant (RP) and acclimated plant (AP) were compared to the mother plant (MP). In addition, the genetic stability of AP was assessed by using inter-simple sequence repeats (ISSR) markers. The highest number of lateral branches (4.25) was obtained from the medium with 3 mg/mL kinetin (KIN), while the highest length of lateral branches (13.25 cm) was achieved on the medium culture fortified with 3 mg/mL thidiazuron (TDZ) and 6-benzylaminopurine (BAP). The highest number of leaves (20.25) and main branch length (12.25 cm) were obtained from the medium containing 3 mg/mL TDZ. The highest number of roots (46.25) and root length (2.25 cm) was measured from the medium fortified with 1 mg/mL indole-3-butyric acid (IBA) and 0.6 mg/mL indole-3-acetic acid (IAA), respectively. RP was successfully acclimated (85%) in vivo. Molecular analysis showed that the AP was true to the type of the MP. cis-Sabinene hydrate (26.8–57.7), 1,8-cineole (6.2–24.1), 4aα,7β,7aα-nepetalactone (4.1–12.3), and terpinene-4-ol (3.2–15.0) were the major essential oils compounds. The studied samples contained rosmarinic acid (2.55–5.97 mg/g DW), cichoric acid (1.68–12.7 mg/g DW), chlorogenic acid (1.91–64.21 mg/g DW), rutin (0.59–1.09 mg/g DW), apigenin (0.52–0.72 mg/g DW), betulinic acid (0.17–2.20 mg g DW), oleanolic acid (0.84–5.37 mg/g DW) and ursolic acid (3.46–15.70 mg/g DW). Acclimated plant exhibited the highest antioxidant activity (IC50 = 196.4 μg/mL), while the methanolic extract of MP displayed the highest antibacterial activity (MIC = 8 mg/mL) against Staphylococcus aureus.

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Acknowledgements

The authors thank the Shahid Beheshti University Research Council for financial support of this project. We also wish to thank Dr. Hassan Esmaeili and Dr. Hamid

Ahadi for their kind help in molecular and phytochemical analyses, respectively.

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  1. Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, 1983969411, Iran

    Mahdieh Zamani & Mohammad Hossein Mirjalili

  2. Department of Biology, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, 1983969411, Iran

    Ali Sonboli & Mostafa Goldansaz

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MZ contributed to the conception of the study, in vitro cultures establishment, data extraction, statistical analysis, and writing of the manuscript. AS helped in plant identification, statistical analysis, and editing the manuscript. SMG contributed in plant materials collection, formal analysis, and revising the manuscript. MHM supervised the whole experiments and wrote the manuscript. All authors read and approved the final manuscript.

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12298_2024_1416_MOESM1_ESM.tif

Fig. S1. ISSR marker analysis generated by the primer IS-5, Lane L corresponds to 3 kb DNA ladder; Lane M, DNA from mother plant; Lanes 1–4, DNA from randomly selected acclimated plantlets in the greenhouse (TIF 1063 KB)

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Zamani, M., Sonboli, A., Goldansaz, M. et al. In vitro micropropagation and conservation of endangered medicinal plant Nepeta asterotricha Rech.f. (Lamiaceae): genetic fidelity, phytochemical and biological assessment. Physiol Mol Biol Plants 30, 67–80 (2024). https://doi.org/10.1007/s12298-024-01416-x

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