Direct bioautography is a powerful technique for localization of different classes of compound of biological interest in higher plants directly on a chromatogram. Polyalthia longifolia Benth. & Hook. is (Annonaceae) an avenue tree and its different plant parts are used in ayurvedic medicines as well as home remedies for common ailments. For isolation and characterization of bioactive compounds from the leaf extract of P. longifolia firstly best TLC solvent system was developed. Further resolution of active compounds was done by TLC bioautography assay. TLC fingerprint of petroleum ether extract after derivatization with anisaldehyde sulphuric acid reagent showed a maximum number of separated compounds. TLC bioautography assay of the petroleum ether extract showed that compounds eluted at Rf value 0.4, 0.5, 0.6 and 0.9, exhibited strong antifungal activity against Alternaria solani. The result indicated that the leaves of P. longifolia might be potential source of antifungal agent.
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