characterization of fungal antagonistic bacilli isolated from aerial roots of banyan ficus benghalensis using intactcell malditof mass spectrometry icms aims to characterize fungal antagonistic bacilli isolated from aerial roots of banyan tree and identify the metabolites responsible for their antifungal activitymethods and results seven gram positive endosporeforming rodshaped endophytic bacterial strains exhibiting a broadspectrum antifungal activity were isolated from the surfacesterilized aerial roots of banyan tree the isolates designated as k1 a2 a4 and a12 were identified as bacillus subtilis whereas isolates a11 and a13 were identified as bacillus amyloliquefaciens using biolog microbial identification system the antifungal lipopeptides surfactins iturins and fengycins with masses varying in the range from mz 900 to mz 1550 could be detected using intactcell malditof mass spectrometry icms on the basis of mass spectral and carbon source utilization profile all seven endophytes could be distinguished from each other furthermore icms analysis revealed higher extent of heterogeneity among iturins and fengycins produced by b subtilis k1 correlating well with its higher antifungal activity in comparison with other isolatesconclusion seven fungal antagonistic bacilli were isolated from aerial roots of banyan tree exhibiting broad spectrum of antifungal activity among which b subtilis k1 isolate was found to be most potent the icms analysis revealed that all these isolates produced cyclic lipopeptides belonging to surfactin iturin and fengycin families and exhibited varying degree of heterogeneitysignificance and impact of the study the endophytes are considered as a potential source of novel bioactive metabolites and this study describes the potent fungal antagonistic bacilli from aerial roots of banyan tree the isolates described in this study have a prospective application as biocontrol agents also icms analysis described in this study for characterization of antifungal metabolites produced by banyan endophytic bacilli may be used as a high throughput tool for screening of microbes producing novel cyclic lipopeptides