Isolation and characterization of indigenous endophytic bacteria associated with leaves of switchgrass (Panicum virgatum L.) cultivars

Abstract

To isolate and characterize indigenous bacterial endophytes from cultivars of switchgrass and study their antimicrobial and growth promoting potential. The diversity, molecular and biochemical characterizations of indigenous and culturable bacterial endophytes residing in leaves of switchgrass have not been studied previously. This study describes the characterization of 31 bacterial endophytes from three switchgrass cutlivars: Cave-in Rock, Blue Jacket and Tecumseh. Molecular and phylogenetic analysis based on the 16S rRNA sequence grouped the endophytes into eight different taxa that shared high homology of 98-99% with other known sequences. Bacterial endophytes were identified as Microbacterium testaceum, Curtobacterium flaccumfaciens, Bacillus subtilis and Bacillus pumilus, Pseudomonas fluorescens, Sphingomonas parapaucimobilis, Serratia sp. and Pantoea ananatis. Some endophytes were detected in switchgrass seeds and in plants that originated from seeds collected a year earlier, confirming vertical transmission to the next generation of the host. Selected endophytes produced cellulases and were capable of solubilizing inorganic phosphorus. Analysis of cell-free culture filtrate of selected strains using direct infusion orbitrap mass spectrometry confirmed the presence of several well-characterized lipopeptide toxins and phytohormones. Re-inoculation of the roots of switchgrass seedlings with endophytes singly or combined confirmed their migration to the upper aerial parts of the plant. Our findings suggest that switchgrass leaves harbour a diversity of bacterial endophytes, some of which could potentially be applied as growth promoting bacteria. This is the first report on the characterization of indigenous bacterial endophytes and their potential use as biofertilizers.