Identification of endophytic bacteria in medicinal plants and their antifungal activities against food spoilage fungi

Abstract

Endophytes are fungi or bacteria living in the intracellular parts of the plants. In this study, 12 medicinal plants were examined for the presence of endophytic bacteria. Antifungal activities of the isolates were determined against Aspergillus flavus PTCC 5006, Penicillium citrinum PTCC 5304, Aspergillus fumigatus PTCC 5006, Fusarium oxysporum MTCC 284, and Rhizopus stolonifer. In order to determine the metabolites characteristics of these isolates, catalase and proteolytic enzyme treatments were assessed. Moreover, approximate molecular weights of the antifungal substance were measured by fractionation method and the volatile compounds were determined by using GC-mass spectroscopy. Finally, 16s rRNA gene sequencing confirmed the strain of the bacteria. Twenty-one endophytic bacteria, out of a total of 82 isolates, showed antifungal activities against all five spoilage fungi. The results of the PCR assay revealed two species: Bacillus pumilus and B. safensis. Proteolytic enzyme activities and the fractionation of the supernatants proved that more than one compound was responsible for the antifungal activities. This compound could be proteins, peptides, and other low-molecular compounds, such as Butanal, 3-methyl-, Propene, 2-butene, 2-heptanone, 6-methyl-5-methylene-, and 6-oxabicyclo[3.1.0] hexane, which all were identified in the headspace of the GC-mass spectroscopy.