In vitro propagation and phytochemical screening of three important ethno-medico-botanicals used by the ethnic communities of Purulia district, West Bengal, India

Abstract

Background: In order to explore the traditional medicine practised by the ethnic communities residing in the Purulia district of West Bengal, India, a quantitative ethnobiological approach was adopted. Three medicinal plants viz. Bacopa monnieri, Aristolochia indica and Rauvolfia serpentina were selected, micropropagted, assessed for clonal fidelity and phytochemical markers.

Materials and methods: Ethnobiological surveys were conducted by interviewing a number of informants or traditional healers with the help of a semi-structured questionnaire. The survey included questions on botanical ingredients and additives, vernacular names of the plants and animals, methods of preparation and administration and restrictions during medications. Additional quantitative indices such as use value, informant’s consensus factor and fidelity level were used for data analysis. On the basis of use value three plants viz. B. monnieri, A. indica and R. sepentina were selected and micropropagted in presence of various plant growth regulators (PGRs) and/or polyamines (PAs). Clonal fidelity of the three micropropagated plants were assessed by RAPD and ISSR based markers. Further, the plants were analyzed by validated HPTLC methods for the presence of marker compounds such as bacoside A, aristolochic acid I, reserpine, ajmalicine and stigmasterol.

Results: A number of folkloric use of botanicals were reported by the traditional healers. High frequency in-vitro multiplication and regeneration in the three selected plants were achieved via use of PGRs and/or PAs. Clonal fidelity assessment of micropropagated plants using two markers (RAPD and ISSR) systems revealed high genetic homogeneity and low level of polymorphism. Comparative HPTLC analysis of bacoside A, aristolochic acid I and reserpine, ajmalicine and stigmasterol content in natural and in-vitro grown B. monnieri, A. indica and R. serpentina respectively indicated higher concentrations of bioactive phytochemicals in tissue-culture raised plants.

Conclusion: Possible synergistic interactions among phytochemicals and additives were indicated to explain enhanced therapeutic efficacy of mixed herbal formulations. Moreover, in-vitro high frequency multiplication and regeneration from various explants may serve as exciting culture material for propagation and conservation of this valuable medicinal plant. HPTLC studies clearly indicated that in-vitro raised plant samples can be used as excellent resource for qualitative and quantitative estimation of high value pharmaceuticals.