Phytochemical Composition and Detection of Novel Bioactives in Anther Callus of Catharanthus roseus L.

Abstract

Catharanthus roseus L. (G.) Don is the most widely studied plant because of its high pharmacological value. In vitro culture uses various plant parts such as leaves, nodes, internodes and roots for inducing callus and subsequent plant regeneration in C. roseus. However, till now, little work has been conducted on anther tissue using plant tissue culture techniques. Therefore, the aim of this work is to establish a protocol for in vitro induction of callus by utilizing anthers as explants in MS (Murashige and Skoog) medium fortified with different concentrations and combinations of PGRs. The best callusing medium contains high α-naphthalene acetic acid (NAA) and low kinetin (Kn) concentrations showing a callusing frequency of 86.6%. SEM–EDX analysis was carried out to compare the elemental distribution on the surfaces of anther and anther-derived calli, and the two were noted to be nearly identical in their elemental composition. Gas chromatography–mass spectrometry (GC–MS) analysis of methanol extracts of anther and anther-derived calli was conducted, which revealed the presence of a wide range of phytocompounds. Some of them are ajmalicine, vindolinine, coronaridine, squalene, pleiocarpamine, stigmasterol, etc. More importantly, about 17 compounds are exclusively present in anther-derived callus (not in anther) of Catharanthus. The ploidy status of anther-derived callus was examined via flow cytometry (FCM), and it was estimated to be 0.76 pg, showing the haploid nature of callus. The present work therefore represents an efficient way to produce high-value medicinal compounds from anther callus in a lesser period of time on a larger scale.

Synthesis and Accumulation of Phytocompounds in Field-, Tissue-Culture Grown (Stress) Root Tissues and Simultaneous Defense Response Activity in Glycyrrhiza glabra L.

“…Previously, Vijayalakshmi and Shourie [39] reported the presence of variety of phytochemicals in leaf, stem, and leaf derived callus in G. glabra. The identification and quantification of bioactive compounds in various plant parts (root, leaf, stem, anther, callus) by GC-MS have recently been described in different plant varieties like Tanacetum sinaicum [40], Catharanthus roseus L. [41] and Amomum nilgiricums [42]. A wide number of therapeutically active phytoconstituents have been detected in G. glabra samples.…”

Elicitation Induced α-Amyrin Synthesis in Tylophora indica In Vitro Cultures and Comparative Phytochemical Analyses of In Vivo and Micropropagated Plants

“…Phytochemical characterization is an immensely valuable step in detecting and quantifying novel bioactive compounds from in vitro-derived tissues (callus, somatic embryo and other plant parts). The application of GC–MS-based chemical profiling for the recognition and documentation of a diverse array of phytochemicals has been exemplified in many scientific reports [ 32 , 33 , 34 ]. In the present study, the GC–MS-based comparative analysis of the metabolites of tissue culture and field-grown (mother) plants was conducted in order to obtain the phytochemical profiles of two alternative sources of T. indica plants.…”

In Vitro Propagation and Phytochemical Composition of Centratherum punctatum Cass—A Medicinal Plant

“…Similar to earlier work, the methanolic extracts of the in vitro-regenerated plant parts-leaf, callus, and root--were separately evaluated through GC-MS in order to detect the active compounds present in the extracts [17,42,43]. In the leaf, 37 phytochemical compounds were isolated, 57 were detected in the callus, and 23 bioactive compounds were found in the root extract.…”

“…After the establishment of a plant cell culture, the researchers tried to analyze the extract in order to identify potential bioactive molecules present in it using GC-MS [2,16]. GC-MS is widely used in various fields like chemistry, biology, pharmacology, and environmental science to examine complex mixtures and to identify unknown compounds [17]. The technology makes it possible to separate a mixture of compounds into individual components using gas chromatography, followed by the analysis of separated components using mass spectrometry, which may detect the ionized fragments.…”