Abstract
Objective: The aim of the present study was to develop a callus induction protocol and comparative study of therapeutic phytochemicals present in in vivo leaf and in vitro callus extracts through Gas Chromatography-Mass Spectrometry analysis.Methods: Murashige and Skoog media was used as culture media for callus induction. In vitro callus induction protocol was developed by studying the effects of various plant growth regulators like auxin, 2, 4-D (2,4-dichlorophenoxyacetic acid), NAA (naphthalic acetic acid), alone and in combination with cytokinin BAP (benzyl aminopurine), on leaf and stem explants. The GC-MS analysis of Ampelocissus latifolia was carried out on Shimadzu QP-2010 plus with thermal desorption system TD 20 to study the phytochemical profile.Results: In vitro callus induction protocol was developed for the plant and callusing was done from leaf and stem explants of Ampelocissus latifolia. The best result for callus induction was obtained using leaf explant, and callus production were maximum in Murashige and Skoog medium fortified with BAP (0.5 mg/l) and NAA (1.0 mg/l). Major compounds identified in the GC-MS analysis were Campesterol, Stigmasterol, Beta-Sitosterol, Docosanol, Dodecanoic acid, etc., in in vitro extract and Beta Sitosterol, Tocopherol, Squalene, Bergamot oil, Margarinic acid, Hexadecanoic acid, etc., in in vivo extract. The different active phytochemicals identified have been found to possess a wide range of biological activities, thus this analysis forms a basis for the biological characterization and importance of the compounds identified for human benefits.Conclusion: This is the first report on callus induction in Ampelocissus latifolia. From the results obtained through the in vitro callus induction and its comparative GCMS analysis with in vivo extract, it is revealed that Ampelocissus latifolia contains various bioactive compounds that are of importance for phytopharmaceutical uses. The GCMS analysis revealed that the amount of Beta-sitosterol and 5-Hydroxymethylfurfural (HMF) was very high in in vitro extract as compared to in vivo extract.
Highlights
From ancient times, medicinal plants have been used extensively for their tremendous healing properties and health benefits
Gas Chromatography-Mass Spectrometry (GC–MS) has been applied unambiguously to identify the structures of different phytoconstituents in plant extracts and biological samples with great success, so further the methanolic leaf and callus extracts were subjected to evaluation for Gas Chromatography-Mass Spectrophotometry analysis to study the phytochemical profile
In vitro callus induction protocol was developed for the plant and callusing was done from leaf and stem explants of Ampelocissus latifolia
Summary
Medicinal plants have been used extensively for their tremendous healing properties and health benefits. Media composition mainly the hormonal balance is an important factor influencing in vitro culture initiation and plant regeneration from explants [3]. Belonging to the family Vitaceae, is an example of ethnomedicinal plant species which is the source of natural dye and a range of traditional medicines that cure various diseases[4]. It is a large herbaceous climber, with a tuberous rootstock. The phytochemical screening may be useful for the detection and isolation of various bioactive compounds which subsequently may lead to drug discovery and development from these plants in vitro
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