BIOTECHNOLOGICAL APPROACHES FOR THE PRODUCTION OF PLUMBAGIN FROM PLUMBAGO ZEYLANICA

Abstract

Demand for medicinal plants has drastically enhanced due to the presence of therapeutically important compounds and is continuously rising in national and international markets. Exploring elite accession among numerous accessions, collected from different locations is an alternative approach to satisfy the increase demand of medicinal plants. Biotechnological approaches are reliable source for production of therapeutically important compound. It also provides long-term utilization of plants. Plumbago zeylanica, a pharmaceutically important medicinal plant has been explored in the present study. In vitro shoot culture was established for five accessions of Plumbago zeylanica. Four different plant tissue culture media, three different carbon sources and three different nitrogen sources were tested for five accessions of P. zeylanica to evaluate optimum culture condition based on growth. The accession which showed maximum shoots number was chosen for further investigation. Accession-based study of in vitro shoot culture showed that accession number IC 524441 is one of the elite accessions and chosen for further studies. Adventitious root suspension culture was explored for enhanced production of plumbagin. Optimization of adventitious root suspension culture showed that highest plumbagin production was obtained in ½ strength MS liquid media having 3% sucrose with 2 g/L of inoculum density. Further elicitation with yeast extract (150 mg/L) increases threefold plumbagin production as compared to control one and highest plumbagin production was 90.96±0.51 µg/mL. Further cell suspension culture was also explored for enhanced production of plumbagin. Optimization of cell suspension culture showed that MS medium having 1 mg/L NAA with 3 g/L inoculum density and 150mg/l yeast extract at pH 5.8 was optimal for plumbagin production. Maximum plumbagin production was enhanced up to 3.3 times as compared to control one and maximum amount of plumbagin production was 83.30±0.18 µg/mL. Biochemical analysis of thirteen different accessions of Plumbago zeylanica were performed where concentration of therapeutically important compounds such as total plumbagin, total flavonoids content, total phenol content, total tannin content and antioxidant activity were evaluated and results showed that IC-524441 is an elite accession. Same thirteen accessions were assessed for genetic diversity analysis using CBDP and SCoT marker. Genetically diverse accessions can be utilized by plant breeders for the generation of elite accessions which have high quantity of pharmaceutically important secondary metabolites. Further residual plant material of P. zeylanica was used for silver nanoparticles synthesis and its application in antibacterial and dye degradation were investigated. Biochar was also derived from residual shoot and root culture and its application in removal of cadmium and chromium were studied. Role of plumbagin against different cancer receptor using in silico method was also evaluated. The ligand plumbagin was docked against three different cancer receptors i.e. COX-2, EGFR and TACE to evaluate its potential effect on different cancer. In-silico studies showed that plumbagin is a potential therapeutic agent for treatment of cancer and same can be established through in vivo studies and clinical trials to confirm its efficiency in patients.