Abstract
Piper longum L. is a well known spice plant belonging to the family Piperaceae with high pharmacognosy potential, but it is becoming threatened due to overexploitation. Thus, this investigation aims to standardize a cost effective protocol for in vitro propagation of this economically important plant. Internodal segments were used as explant for callogenesis in Murashige and Skoog medium with 3% sucrose and 0.8% agar, with NAA or 2,4 D. Optimum callus induction was observed in MS medium with 5.0 mg/L NAA. Calli were subcultured on shoot regeneration media containing different concentrations of cytokinin (KIN/BAP) along with 0.1 mg/L NAA. Best shoot regeneration was obtained on MS media supplemented with 2.0 mg/L KIN and 0.1 mg/L NAA. Induced shoots were rooted in either NAA or IBA and highest rooting was induced in MS medium enriched with 0.5 mg/L NAA. Rooted plantlets were acclimatized and 88% of hardened plants survived. Field emission scanning electron microscopic showed that regeneration from callus had occurred by somatic embryogenesis. A comparative study on identification and quantification of piperine (the chief alkaloid of the cultivar) were done from root and fruit of both in vitro and in vivo grown plants through Reverse Phase-High Performance Liquid Chromatography method. In vitro grown fruit was found to have the maximum amount of piperine.
Key message
For the first time, a comparative study in identification and quantification of piperine was done from both in vitro and in vivo grown Piper longum through RP-HPLC method.
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Abbreviations
- MS:
-
Murashige and Skoog
- NAA:
-
1-Napthaleneacetic acid
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- KIN:
-
Kinetin
- BAP:
-
N6-Benzylaminopurine
- IBA:
-
Indole-3-butyric acid
- mg/L:
-
Milligram/Litre
- FE-SEM:
-
Field emission scanning electron microscope
- RP-HPLC:
-
Reverse Phase-High Performance Liquid Chromatography
- Rt :
-
Retention time
- ANOVA:
-
Analysis of variance
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Acknowledgements
Authors are thankful to the University of Burdwan, Department of Biotechnology for providing us the infrastructural facilities and opportunity to done this research work. Authors also want to thank and convey their sincere gratitutude to Mr. Kaushik Sarkar (Technical assistant of Dept. of Biotechnology, The university of Burdwan, WB, India) for helping in HPLC analysis.
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The experiment was designed by MC and Dr. IC Collection of plant, sample preparation and whole research work was executed and statistically analyzed by MC. Dr. SC monitored the research significantly. This experimental work is a part of Ph.D. thesis of MC. All authors read the manuscript, revised critically and approved the final version.
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Chatterjee, M., Chatterjee, S. & Chandra, I. In vitro regeneration of Piper longum L. and comparative RP-HPLC analysis of piperine production of in vitro and in vivo grown plants. Plant Cell Tiss Organ Cult 149, 205–212 (2022). https://doi.org/10.1007/s11240-022-02237-0
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DOI: https://doi.org/10.1007/s11240-022-02237-0