Cell wall polysaccharides of endophytic Pseudomonas putida elicit defense against rice blast disease.

Abstract

To characterize the functional role of extracellular polysaccharides and lipopolysaccharide extracted from endophytic Pseudomonas putida BP25 (PpBP25) against rice blast. We profiled the transcriptome of endobacterized rice seedlings using RNA-seq. Fluorescence imaging of interaction between Magnaporthe:: gfp and P. putida:: mCherry was performed on rice-phylloplane using Confocal Laser Scanning Microscopy (CLSM). Microbial polysaccharides, EPS and LPS extracted from PpBP25 were characterized using Fourier-transform infrared-spectroscopic analysis (FTIR). Biochemical assays, and gene expression analysis were conducted on EPS and LPS-treated rice seedlings. A detached-leaf assay was designed to test the blasticidal-effect of bacterial-endophyte, EPS and LPSon rice-phylloplane.PpBP25elicited defense in rice with a consequently altered seedling phenotype. Rice cultivar, Pusa Basmati-1, colonized by PpBP25 showed an altered transcriptome profile displaying a total of 110-downregulated and 68-upregulated genes (p<0.005) representing growth/development and defense pathways, respectively. CLSM of PpBP25 bacterized phylloplane showed reduced conidial-germination and mycelial-biomass of M. oryzae. To decipher the elicitor role of polysaccharides, we purified and characterized EPS and LPS using FTIR. Rice treated with the EPS and LPS showed root-growth inhibition -the phenotype of MAMP-triggered immunity. While the EPS showed blast suppressive activity at 1-20mg mL-1 (79.80-86.87% reduction over control), the LPS exhibited 78.0-79.8% reduction at 20-200µg mL-1on rice. Polysaccharides treated seedling showed elevated activities of peroxidase and polyphenol-oxidase activities, and total-phenols content. Treated plantlets showed up regulation of OsPR1.1, OsPR3, OsGLP3-3 OsZFP179, and Oshox24 as well as down regulation of OsACS6. We showed that P. putida and its cell-wall associated polysaccharides could elicit defense against rice blast.