Optimization of Sterilization Parameters for Isolation of Endophytes from Allium sativum and Exploring its Antibacterial Activity

Abstract

Endophytic bacteria reside within the plant cell and are beneficial to it in a number of ways like growth, protection to environmental conditions and sustainability in favor of the hosts. Throughout the ages, Garlic (Allium sativum L.) has legendary therapeutic importance. The important step in endophyte isolation is the removal of plant tissue surface microflora, called as surface sterilization. Endophytes from medicinal plants may mimic the compound produced by the host plant and also plays an important role in production of bioactive compound, so it is necessary to isolate endophytes, not the epiphytic microbes. The present investigation was undertaken with an aim to optimize the sterilization parameters viz: time and concentration of ethanol and sodium hypochlorite for isolation of endophytic bacteria as well as exploring their antibacterial activity from Allium sativum. The results revealed that concentration and exposure time of sterilizing agent caused prominent surface sterilization but have negative effect on isolation of endophytes. Experimental results revealed that 70% ethanol (6 min), 2% sodium hypochlorite (1 min) followed by 70% ethanol (30 sec) is effective for surface sterilization of leaf and 70% ethanol (6 min), 3% sodium hypochlorite (1 min) followed by ethanol (30 sec) for bulb of Allium sativum. A total of 86 bacterial endophytes were recovered and screened for antibacterial activity against Enterococcus faecalis, Bacillus cereus ATCC 11778, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Enterobacter, Klebsiella pneumoniae, Salmonella typhi and Proteus. Out of 86 bacterial endophytes, only (9%) endophytes were found to be inhibiting one or all test human pathogen. The findings of the present study suggest that use of optimized sterilization conditions are effective for removing surface bacterial strains without affecting endophytes as well as the Allium sativum plant of the Jaunpur district represents an excellent reservoir of endophytic bacteria and novel source of bioactive compounds. On the other hand different concentration and time of their exposure to different parts of plant is required for proper surface sterilization of that part and isolation of endophytes.