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A viable alternative in vitro system and comparative metabolite profiling of different tissues for the conservation of Ceropegia karulensis

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Abstract

Ceropegia karulensis is an endemic and critically endangered plant of the Western Ghats from India. Exploitation of the tubers and poor regeneration from seed has narrowed distribution and propagation of the species. There is a need to develop in vitro propagation methods for C. karulensis to alleviate these problems. Here, we optimized callus induction, somatic embryogenesis and microtuberization from different seedling explants viz. cotyledonary leaf and root. The environmental scanning electron microscopy was used to observe somatic embryonic origin and their developmental stages. Highest callus proliferation was recorded with 2 µM 6-benzylaminopurine and 1 µM 2,4-dichlorophenoxyacetic acid. Somatic embryos derived from cotyledonary leaf explants were more proliferative than root explants. The combination of 2 µM 6-benzylaminopurine, 2 µM naphthalene acetic acid and 7% sucrose in MS media resulted in highest microtuberization. Further, gas chromatography-mass spectrometry based metabolic profiling was carried out from native wild plants and in vitro callus tissues which identified various phytochemicals such as alkaloids, fatty acids, esters alcohols, etc. Multivariate analysis revealed the chemical disparities, where considerable variations were observed between native wild type and in vitro tissues, but no significant differences were found among in vitro callus from both root and cotyledonary explants. Overall, our results suggested that the production of various secondary metabolites found in C. karulensis was not affected by in vitro propagation and could be utilized in the conservation strategies for this plant.

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Abbreviations

BAP:

6-benzylaminopurine

2,4-D:

2,4-dichlorophenoxyacetic acid

NAA:

Naphthalene acetic acid

MS:

Murashige and Skoog’s medium

PGR(s):

Plant growth regulator(s)

HF:

Hormone-free

SEs:

Somatic embryos

ESEM:

Environmental scanning electron microscopy

GC-MS:

Gas chromatography-mass spectrometry

GC-FID:

Gas chromatography-flame ioinzation detector

CAL:

Central National Herbarium

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Acknowledgements

MP would like to thank the CSIR-National Chemical Laboratory, Pune (Maharashtra) India, for providing laboratory facilities. Financial assistance from Department of Science and Technology (Project No. SB/YS/LS-266/2013) and from CSIR network project under XII five-year plan (BSC0124), Government of India, New Delhi, is gratefully acknowledged.

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  1. Ramesha H. Jayaramaiah

    Present address: Department of Plant Bioscience, Life and Industry Convergence Research Institute, Pusan National University, Miryang, 627–707, South Korea

Authors and Affiliations

  1. Plant Molecular Biology Laboratory, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune, Maharashtra, 411008, India

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

  2. Biospheres, Eshwari, 52/403, Lakshminagar, Parvati, Pune, Maharashtra, 411 009, India

    Sachin A. Punekar

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Contributions

APG and MP conceived and planned the work; SAP identified the species; all authors were involved in all field work; MP and RHJ designed and performed the experiments and analyzed the data; all authors wrote and approved the final manuscript.

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Correspondence to Ashok P. Giri.

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Communicated by Ali R. Alan.

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Pandey, M., Jayaramaiah, R.H., Dholakia, B.B. et al. A viable alternative in vitro system and comparative metabolite profiling of different tissues for the conservation of Ceropegia karulensis . Plant Cell Tiss Organ Cult 131, 391–405 (2017). https://doi.org/10.1007/s11240-017-1292-6

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