Hydrocarbon-derived pollutants are becoming one of the most concerning ecological issues. Thus, there is a need to investigate and develop innovative, low-cost, eco-friendly, and fast techniques to reduce and/or eliminate pollutants using biological agents. The study was conducted to isolate, characterize, and identify potential diesel-degrading bacteria. Samples were collected from flower farms, lakeshores, old aged garages, asphalt, and bitumen soils and spread on selective medium (Bushnell Haas mineral salt agar) containing diesel as the growth substrate. The isolates were characterized based on their growth patterns using optical density measurement, biochemical tests, and gravimetric analysis and identified using the Biolog database and 16S rRNA gene sequencing techniques. Subsequently, six diesel degraders were identified and belong to Pseudomonas, Providencia, Roseomonas, Stenotrophomonas, Achromobacter, and Bacillus. Among these, based on gravimetric analysis, the three potent isolates AAUW23, AAUG11, and AAUG36 achieved 84%, 83.4%, and 83% diesel degradation efficiency, respectively, in 15 days. Consequently, the partial 16S rRNA gene sequences revealed that the two most potent bacterial strains (AAUW23 and AAUG11) were Pseudomonas aeruginosa, while AAUG36 was Bacillus subtilis. This study demonstrated that bacterial species isolated from hydrocarbon-contaminated and/or uncontaminated environments could be optimized to be used as potential bioremediation agents for diesel removal.
Grapevine is an economically important crop throughout the world. However commonly cultivated species Vitis vinifera is susceptible to powdery mildew fungi. The objective of this study was to analyze the promoter of VvNAC36 transcription factor gene, which was induced by powdery mildew infection, as it was shown by previous microarray results. Analysis of VvNAC36 transcription factor promoter allows identifying the important cis-elements, which are responsible for gene regulation during infection. Knowing that the responsible cis-element of the gene is crucial for further study, the role of this gene is in powdery mildew infected grapevine. The study was performed using circumspect methodology. To confirm the presence of transgene in transgenic plants the PCR and glufosinate total herbicide application was used. The transplanted transgenic seedlings were infected with Golovinomyces orontii. To detect the GUS reporter gene expression, histochemical GUS staining and spectrophotometric assays were applied in each deletion variant. Finally the promoter sequence was analyzed by the PLACE (a database for PLAntCisacting Elements) program for identifying cis-elements. The results of the study showed significant difference in mock treated and induced expression in transgenic plants transformed with VvNAC36 promoter segments of different lengths, fused to GUS reporter gene. In basal expression level, the deletion variants 1178 bp and 257 bp differed significantly. Powdery mildew infection significantly increased the expression in all promoter variants, except the two shortest fragments. Based on the promoter motif analysis, it can be concluded, that the GT1-box (GAAAAA) supposed to have crucial role in regulating of VvNAC36 transcription factor gene during powdery mildew infection.
Kerosene is widely used in Ethiopia as a household fuel (for lighting and heating), as a solvent in paint and grease, and as a lubricant in glass cutting. It causes environmental pollution and escorts to loss of ecological functioning and health problems. Therefore, this research was designed to isolate, identify, and characterize indigenous kerosene-degrading bacteria that are effective in cleaning ecological units that have been contaminated by kerosene. Soil samples were collected from hydrocarbon-contaminated sites (flower farms, garages, and old-aged asphalt roads) and spread-plated on mineral salt medium (Bushnell Hass Mineral Salts Agar Medium: BHMS), which consists of kerosene as the only carbon source. Seven kerosene-degrading bacterial species were isolated, 2 from flower farms, 3 from garage areas, and 2 from asphalt areas. Three genera from hydrocarbon-contaminated sites were identified, including Pseudomonas, Bacillus, and Acinetobacter using biochemical characterization and the Biolog database. Growth studies in the presence of various concentrations of kerosene (1% and 3% v/v) showed that the bacterial isolates could metabolize kerosene as energy and biomass. Thereby, a gravimetric study was performed on bacterial strains that proliferated well on a BHMS medium with kerosene. Remarkably, bacterial isolates were able to degrade 5% kerosene from 57.2% to 91% in 15 days. Moreover, 2 of the most potent isolates, AUG2 and AUG1, resulted in 85% and 91% kerosene degradation, respectively, when allowed to grow on a medium containing kerosene. In addition, 16S rRNA gene analysis indicated that strain AAUG1 belonged to Bacillus tequilensis, whereas isolate AAUG showed the highest similarity to Bacillus subtilis. Therefore, these indigenous bacterial isolates have the potential to be applied for kerosene removal from hydrocarbon-contaminated sites and the development of remediation approaches.
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