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CLONING OF RING-HYDROXYLATING OXYGENASE GENES AND SCREENING OF BACTERIA FOR PHYTOREMEDIATION

2017
Stockmans, Anneleen
Professionele bachelor in de agro- en biotechnologie

Abstract :
For phyto- and phyllo-remediation, it is important to know if there is degradation of the pollutant present in the soil or in the air. To solve this problem, a goal of this study is to optimize a Quantitative Polymerase Chain Reaction (qPCR). Through this molecular method, it would be easy to tell if there are aromatic pollutant degrading microorganisms present or not in the soil or phylloplane. By measuring the abundance of these ring-hydroxylating oxygenase genes, a correlation between the presence of these genes and the concentration of pollutants in the environment can be made. This allows to estimate the degree of phytoremediation taking place at the polluted site. The tested genes here were alkane monooxygenase, phenol hydroxylase and naphthalene dioxygenase. Also, the total population of bacteria and fungi was determined. The genes were Polymerase Chain Reaction (PCR) - amplified and cloned from bacteria. These cloned genes were used to set up the standard curves for the qPCR. Second, bacteria isolated out of polluted soil, were screened on aromatic ring-hydroxylating oxygenase (RHO) genes including alkane monooxygenase, phenol hydroxylase, naphthalene dioxygenase, catechol dioxygenase, xylene monooxygenase, toluene dioxygenase and biphenyl dioxygenase. From this, the positive bacteria can be further used in phytoremediation. A PCR amplifies those genes, so that the presence in the bacteria can be determined. Lastly, the new bacterium Acinetobacter calcoaceticus strain Gkorezis (GK) 2, was screened for his degradative and plant growth promoting potential to be used in phytoremediation by means of genome annotation. The tests and optimizations revealed successful new insights. Naphthalene dioxygenase-, phenol hydroxylase- and alkane monooxygenase genes were successfully cloned and the qPCR was optimized. There are significant correlations found between pollutant concentrations and the presence of degradative genes. Also, new aromatic-eating bacteria were isolated and interesting degradation pathways in strain GK2 were found. Possible soil and air pollutant remediation solutions must be further investigated, because of the positive effects these types of plant-microbe interactions have on the environment.

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If you want to cite this thesis in your own thesis, paper, or report, use this format (APA):

Stockmans, A. (2017). CLONING OF RING-HYDROXYLATING OXYGENASE GENES AND SCREENING OF BACTERIA FOR PHYTOREMEDIATION. Unpublished thesis, Hogeschool PXL, PXL-Tech.
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