Development of random amplified polymorphic DNA markers for authentication of Baliospermum montanum Willd. leaf with its pharmacognostical evaluation

Abstract

Introduction: Baliospermum montanum Willd. (Euphorbiaceae) is an undershrub, distributed in outer range of Himalayas from Kashmir to Assam and in moist deciduous forests elsewhere in India. Leaves, seeds, and roots of this plant are being used for medicinal purpose. Aim: The aim of this study was to carry out detailed pharmacognostical study of B. montanum Willd. (Euphorbiaceae) leaf and its molecular characterization by random amplified polymorphic DNA (RAPD) markers. Materials and Methods: Fresh young leaves of Baliospermum montanum Willd, after proper botanical authentication, were subjected for macro and microscopic study and were also used for molecular characterization and DNA fingerprints, by standard and most convenient RAPD method. Results and Discussion: Microscopic study of leaf shows the presence of single layer of epidermis made up of single-layered barrel-shaped cells interrupted by unicellualar trichomes. Mesophyll made up of single-layered compactly arranged cells filled with chlorophyll pigment. Transverse section of petiole shows single-layered barrel-shaped epidermal cells without any intracellular space, hypodermis consists of 8-10 layers of collenchymatous cells filled with rosette crystals and oil globules, reduced cortex filled with chlorophyll pigments. Vascular bundles consist of radially and circularly arranged open and collateral phloems. DNA RAPD analysis reveals that all the primers showed amplification. In primer 1, range of band size was observed from 400 to 1200 bp; in primer 2, band size was observed at 250 bp to around 1500 bp.Conclusion: These microscopic observations and the unique bright and light bands obtained in polymerase chain reaction amplification could serve as a measure for authentication and standardization of the plant.