Aim: Plant secondary metabolites are the alternatives for the control of the antibiotic resistant pathogens which are highly infectious to human and plants. With respect to these aspects present work was carried out with phytochemical studies and antibacterial screening of Pongamia pinnata Pierre.
 Materials: The Preliminary phytochemical screening and HPTLC studies were performed with methanolic extracts of seeds and callus. The extracts of the seeds and callus was developed in the mobile phase of toluene: ethyl acetate (90:10) using standard procedures and scanned under UV at 254 nm, 366 nm and visible light at 540 nm.Further, the extracts of seeds and callus were made in diethyl ether and methanol used for in vitro antibacterial activity. It was performed against multi drug resistance organisms such as Klebsiella pneumoniae, Escherichia coli, Bacillus subtilis and Staphylococcus aureus. The organic extracts at a concentrations of 0.02 mg/ml - 0.1 mg/ml were taken to study the inhibition properties.
 Results: The Pongamia pinnata Pierre HPTLC fingerprint analysis from the extracts showed a wide range of secondary metabolites. The methanolic extracts of seed and callus showed significant inhibition zones.
 Conclusions: The study reveals the new antimicrobial agent in the form of Pongamia pinnata callus extracted with methanol will be definitely an alternative to synthetic drugs or antibiotic agents.
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