Tropolone, a biotoxin produced by the agricultural pathogen Burkholderia plantarii, exerts cytotoxicity toward a wide array of biota. However, due to the lack of quantitative and qualitative approach, both B. plantarii occurrence and tropolone contamination in agricultural environments remain poorly understood. Here, we presented a sensitive and reliable method for detection of B. plantarii in artificial, plant, and environmental matrices by tropolone-targeted gas chromatography-triple-quadrupole tandem mass spectrometry analysis. Limits of detection for B. plantarii and tropolone were 10 colony-forming units (CFU)/mL and 0.017 μg/kg, respectively. In a series of simulation trials, we found that B. plantarii from 10 to 10 CFU/mL produced tropolone between 0.006 and 107.8 mg/kg in a cell-population-dependent manner, regardless of habitat. Correlation analysis clarified a reliable reflection of B. plantarii density by tropolone level with R values from 0.9201 to 0.9756 ( p < 0.01). Through a nationwide pilot study conducted in China, tropolone contamination was observed at 0.014-0.157 mg/kg in paddy soil and rice grains, and subsequent redundancy analysis revealed soil organic matter to be a dominant environmental factor, having a positive correlation with tropolone contamination. In this context, our results imply that potential ecological and dietary risks posed by long-term exposure to trace levels of tropolone contamination are of concern.
Seed borne bacterial pathogens cause severe yield loss and biotoxin contamination in rice, leading to an increasing concern on the global food supply and environmental safety. Plant native microbes play an important role in defending against diseases, but their actions are often influenced by the chemical fungicides applied in the field. Here, Bacillus licheniformis mmj was isolated from rice spikelet, which uniquely showed not only the fungicide-responsiveness but also the broad-spectrum antimicrobial activity against major rice bacterial pathogens including Xanthomonas oryzae pv. oryzae, Burkholderia plantarii and Burkholderia glumae. To understand the hallmark underlying the environmental adaption and antimicrobial activity of B. licheniformis mmj, the genome sequence was determined by SMRT and subjected to bioinformatics analysis. Genome sequence analysis enabled to identification of a set of the antimicrobial-resistance and antibacterial activity genes together with an array of harsh environment-adaptive genes. Moreover, B. licheniformis mmj metabolites were analyzed with gas chromatography coupled to triple quadrupole mass spectrometry, and the volatile components that were linked with the antimicrobial activity were preliminarily profiled. Collectively, the present findings reveal the genomic and metabolic landscapes underlying the fungicide-responsive B. licheniformis, which offers a new opportunity to design harsh environment-adaptive biopesticides to cope with the prevalent bacterial phytopathogens.
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