Native strains of Beauveria spp. were evaluated against Spodoptera litura using leaf spray method. Twelve native and one reference strain of B. bassiana were assayed at 1×1010 spores ml-1 to determine LT50 by probit analysis. The least LT50 values of 135.84 and 179.96 hours were recorded in native strains of SGb and MKb followed by Hb, Kb and Bb i.e. LT50 value of 195.49, 201.20 and 235.30. The highest LT50 values of 500.88 and 336.90 hours were recorded in RARST-b and NBAII-b strains. Five effective and promising B. bassiana strains (SGb, MKb, Kb, Bb and Hb) were characterized using rDNA-ITS sequences. All the five strains of B. bassiana showed 100% homology with already submitted B. bassiana strain in NCBI GenBank. Phylogenic dendrogram exhibited B. bassiana strains Bb and Kb as distinct cluster compared to others, where as B. bassiana strains SGb, MKb and Hb formed one cluster, however SGb grouped into same clade with AB 027382 (IFO 4848) and FJ 755242 (CZ 590) which were isolated from China and Japan, respectively. Gene sequences of five native strains were deposited in NCBI Gene Bank, Bethesda, USA with the accession numbers: JX173280.1 (SGb, Bb1), JX313063.1 (MKb, Bb2), JX313064.1 (Kb, Bb3), JX313065.1 (Bb, Bb4) and JX313066.1 (Hb, Bb5).