Simple SummaryBurkholderia contaminans belongs to B. cepacia complex (Bcc), those of which are found in various environmental conditions. In this study, a novel strain AY001 of B. contaminans (AY001) was identified from the rhizosphere soil sample. AY001 showed (i) various plant growth-promoting rhizobacteria (PGPR)-related traits, (ii) antagonistic activity against different plant pathogenic fungi, (iii) suppressive activity against tomato Fusarium wilt disease, (iv) induced systemic acquired resistance (ISR)-triggering activity, and (v) production of various antimicrobial and plant immune-inducing secondary metabolites. These results suggest that AY001 is, indeed, a successful PGPR, and it can be practically used in tomato cultivation to alleviate biotic and abiotic stresses. However, further safety studies on the use of AY001 will be needed to ensure its safe use in the Agricultural system.Plant growth promoting rhizobacteria (PGPR) is not only enhancing plant growth, but also inducing resistance against a broad range of pathogens, thus providing effective strategies to substitute chemical products. In this study, Burkholderia contaminans AY001 (AY001) is isolated based on its broad-spectrum antifungal activity. AY001 not only inhibited fungal pathogen growth in dual culture and culture filtrate assays, but also showed various PGPR traits, such as nitrogen fixation, phosphate solubilization, extracellular protease production, zinc solubilization and indole-3-acetic acid (IAA) biosynthesis activities. Indeed, AY001 treatment significantly enhanced growth of tomato plants and enhanced resistance against two distinct pathogens, F. oxysporum f.sp. lycopersici and Pseudomonas syringae pv. tomato. Real-time qPCR analyses revealed that AY001 treatment induced jasmonic acid/ethylene-dependent defense-related gene expression, suggesting its Induced Systemic Resistance (ISR)-eliciting activity. Gas chromatography–mass spectrometry (GC-MS) analysis of culture filtrate of AY001 revealed production of antimicrobial compounds, including di(2-ethylhexyl) phthalate and pyrrolo [1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl). Taken together, our newly isolated AY001 showed promising PGPR and ISR activities in tomato plants, suggesting its potential use as a biofertilizer and biocontrol agent.