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Combinatorial Approach Through In Vitro Regeneration and Phytochemical Profiling of Ceropegia media (Huber) Ans.: A Potential Way Forward in the Conservation of an Endangered Medicinal Plant from the Western Ghats in India

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Abstract

Ceropegia media is an endemic and endangered plant as its propagation through seeds is unreliable due to low germination, slow growth and seedling decay under natural conditions. Also, tubers of this plant are edible serving as carbohydrate source with medicinal values leading to severe population decline in the natural habitat. To provide a sustainable solution, an efficient in vitro propagation protocol along with phytochemical profiling was developed for C. media. Callus cultures were induced from seedling and wild leaf tissues using the most effective Murashige and Skoog’s (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D; 2 µM) and sucrose (3%). Somatic embryos were acquired on MS medium with 1 µM 6-Benzylaminopurine (BAP) and 1 µM 2,4-D. Conversion into plantlets was attained only from tissue culture-derived seedling leaf (TCDSL) explant. Further, in vitro tuberization was achieved from TCDSL callus with BAP and Naphthalene acetic acid (NAA). AgNO3 as an elicitor had a positive effect on both fresh and dry weights of callus. Successful acclimatization (58%) was attained after two months resulting in normal phenotype in pots. Further, metabolite profiles of ten different tissues from wild and in vitro plants were compared. Total 82 compounds comprising alkaloids, fatty acids, fatty acid ester, steroids, terpenes and hydrocarbons were identified. Overall, results suggested enhanced production of selected metabolites with in vitro propagation and AgNO3, alleviating the problem of unavailability of planting materials. Thus, the current study might offer potential ways for the conservation of such RED enlisted species as C. media.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

AgNO3 :

Silver nitrate

ANOVA:

Analysis of variance

BAP:

6-Benzylaminopurine

BSI:

Botanical Survey of India

ESEM:

Environmental scanning electron microscopy

GC-FID:

Gas chromatography-flame ionization detector

GC–MS:

Gas chromatography-mass spectrometry

IAA:

Indole-3-acetic acid

Kin:

Kinetin

MS:

Murashige and Skoog’s medium

NAA:

Naphthalene acetic acid

PGR:

Plant growth regulator

PLS-DA:

Partial least square-discriminant analysis

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Acknowledgements

Financial assistance to MP from Department of Science and Technology, Government of India, New Delhi (Project No. SB/YS/LS-266/2013), is gratefully acknowledged.

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Authors and Affiliations

  1. Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune, Maharashtra, 411008, India

    Meena Pandey, Bhushan B. Dholakia, Ramesha H. Jayaramaiah & Ashok P. Giri

  2. Biospheres, Eshwari, 52/403, Lakshminagar, Parvati, Pune, Maharashtra, 411009, India

    Sachin A. Punekar

  3. Hawkesbury Institute for the Environment, Western Sydney University, Penrith, NSW, 2751, Australia

    Ramesha H. Jayaramaiah

  4. Indian Institute of Science Education and Research-Pune, Dr. Homi Bhabha Road, Pune, Maharashtra, 411008, India

    Bhushan B. Dholakia

  5. D Y Patil International University, D Y Patil Educational Complex, Sector 29, Nigdi Pradhikaran, Akurdi, Pune, Maharashtra, 411044, India

    Meena Pandey

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  1. Meena Pandey
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Contributions

APG and MP conceived and planned the work; APG supervised the work. SP Identified the species. All the authors were involved in all field work. MP conducted in vitro study, analyzed the data, while MP and RHJ performed GC–MS experiments and analyzed the data. MP prepared figures and initial draft of the manuscript with some inputs from BBD. Subsequently, BBD and APG suggested changes and edited entire manuscript. All authors read and approved the final manuscript.

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Correspondence to Meena Pandey, Bhushan B. Dholakia or Ashok P. Giri.

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Pandey, M., Dholakia, B.B., Jayaramaiah, R.H. et al. Combinatorial Approach Through In Vitro Regeneration and Phytochemical Profiling of Ceropegia media (Huber) Ans.: A Potential Way Forward in the Conservation of an Endangered Medicinal Plant from the Western Ghats in India. J Plant Growth Regul 40, 1139–1151 (2021). https://doi.org/10.1007/s00344-020-10173-6

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