EVALUATION OF ANTIBIOTIC AND BIOCHEMICAL POTENTIAL OF BRYOPHYTES FROM KUMAUN HILLS AND TARAI BELT OF HIMALAYAS Original Article VIDISHA KANDPAL, PREETI CHATURVEDI*, KAVITA NEGI, SHUBHPRIYA GUPTA, ANITA SHARMA Department of Biological Sciences, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar Email: an_priti@yahoo.co.in Received: 07 May 2015 Revised and Accepted: 20 Apr 2016 ABSTRACT Objective: Today, one of the major problems in the treatment of disease is the development of resistance against conventional antibiotics. One way to curb the problem of increasing antibiotic resistance is to use botanicals. Bryophytes, one of the earliest land inhabitants, are generally not known to get affected by any disease in nature owing to their unique chemical constituents. Therefore, the study was aimed to test the efficacy of bryophytes as an alternative to the synthetic drugs by exploring their antimicrobial and biochemical potential. Methods: Antibacterial, biochemical and antioxidant characterization of 2 liverworts, Reboulia hemisphaerica L. (Raddi), Marchantia palmata Reinw., Nees & Blume and one moss species, Hydrogonium gracilantum (Mitt). P. C. Chen was done under laboratory conditions. Results: Both acetone and ethanol extracts of the collected bryophytes inhibited the growth of Escherichia coli, Bacillus cereus, Erwinia chrysanthemi and Pseudomonas aeruginosa on an agar plate. The ethanol extract of H. gracilantum was the most potent inhibitor of E. chrysanthemi followed by ethanol extract of R. hemisphaerica against E. coli. Conclusion: E. chrysanthemi was the most sensitive pathogen to ethanol extract of H. gracilentum while E. coli and B. cereus were inhibited most by ethanol extract of R. hemispherica. However, P. aeruginosa was most sensitive to acetone extracts of M. palmata and H. gracilantum and ethanol extract of R. hemisphaerca. All the plant extracts had moderate content of phenols and flavonoids. The antioxidant activity of corresponding extracts could be related with the total phenol and flavonoid contents. Keywords: Bryophytes, Reboulia hemisphaerica, Marchantia palmata, Hydrogonium gracilantum, Phenols, antioxidants, Flavonoids, Antibacterial © 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) INTRODUCTION The oldest land plants on earth, i.e., “bryophytes” comprise of three phyla: the liverworts (Marchantiophyta), the mosses (Bryophyta) and the hornworts (Anthocerophyta). Despite the rich diversity and unique characteristics of bioactive molecules of bryophytes, their medicinal importance is negligibly explored. Interestingly, the plants are being used in the ethnomedical field from times immemoriall [1]. However, the group has drawn the attention of plant and chemical scientists for application potential only in the last few decades [2, 3]. Pharmacological investigations of bryophytes have proved that the active principles of the plants are very unique and have the remarkable potential of therapeutic applications. Presently, over 400 new compounds have been isolated and characterized for their biochemical and antimicrobial properties. The enzymatic machinery of these plants also has antioxidative property which helps them to withstand the extreme climate and stress conditions particularly desiccation [4]. An important observation about the bryophytes is that these are rarely infected by microorganisms through their habitat, being moisture rich, is usually very prone to microbial attacks. Bryophytes, particularly liverworts, are known to show antibacterial and antifungal activity [5]. Both Reboulia hemisphaerica (Aytoniaceae) and Marchantia palmata used in the present study are thalloid liverworts. Marchantia spp are known to be used in folk remedies for cancer [6]. The moss, Hydrogonium gracilantum grows as a pure or mixed patch on the hills. All these plants used in the present study have been unexplored for their biochemical and antioxidant potential. There is not much information available in the literature on the antioxidative and biochemical potential of these bryophytes. The present study is focused on the preliminary evaluation of phytochemicals, assessment of antioxidative and antibacterial potential of aqua-ethanolic and aqua-acetonic extracts of the three bryophytes viz., Reboulia hemisphaerica, Marchantia palmata and Hydrogonium gracilantum under in vitro conditions for analyzing their potential as a therapeutic source on the basis of plant type and the solvent system used for extraction. MATERIALS AND METHODS Collection of plant material Gametophytes of Reboulia hemisphaerica and Hydrogonium gracilantum were collected from Dwarahat (29.78 °N latitude,79.43°E longitude at an altitude of 1499m asl), Almora distt. and Marchantia palmata was collected from Pantnagar (29.02 °N latitude, 79.30 °E longitude at an alt. of 243m asl) in U. S. Nagar distt. of Uttarakhand, India. Voucher specimens of the bryophytes have been deposited in the herbarium maintained at Department of Biological Sciences. Preparation of plant extract The plants with rhizoids were extensively washed with running tap water, spread on the paper sheet, shade dried, pulverized and extracted by cold percolation (10 g/100 ml) in 80 % of ethanol and acetone solvents. The extracts were filtered and concentrated using rotary evaporator (Biogen). Different concentrations of the crude extract (100, 400, 700 and 1000 μg/ml) were prepared and used for the further study. Microorganisms To test the antibacterial activity of the plant extracts, 4 common pathogenic bacteria were used. The test bacteria viz., Escherichia coli (MTCC 443), Pseudomonas aeruginosa (MTCC 424), and B. cereus (MTCC 430) were procured from Institute of Microbial Technology, Chandigarh. Erwinia chrysanthemi was kindly provided by Department of Microbiology, G. B. Pant University of Agriculture & Technology, Pantnagar. Antibacterial activity Antibacterial activity of all the plant extracts was assessed by agar well diffusion method according to Kumar & Chaudhary [7]. Different concentrations of crude ethanol and acetone extracts (40 µl) were pipetted out into the wells of bacteria seeded nutrient agar International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 6, 2016