ARCHIVES
VOL. 8, ISSUE 1 (2023)
Testing the ability of plasmid DNA content in Rhodotorula mucilaginosa S6 to degradation hydrocarbon compounds and transfer through conjucation
Authors
Sarah I Hayes, Badia A Malla Obaida
Abstract
The current study
included isolation of yeasts from the soils of Bay Hassan oil field in Kirkuk
Governorate and the K1 oil filling station in Erbil Governorate/ Iraq, and it was
diagnosed based on phenotypic, culture and biochemical tests. The results
showed that the yeasts belong to the species: Saccharomyces cerevisiae
and Rhodotorula mucilaginosa. Also, Escherichia. coli bacteria
were obtained from the exit of infants in Ibn Al-Atheer Teaching Hospital in
Mosul/Iraq and was diagnosed based on biochemical tests. The resistance of
isolates was studied for 16 antibiotics and 8 heavy metal ions. The resistance
of R. mucilaginosa S6 and S. cervisiae S3 isolates to 16
antibiotics and 8 heavy metal ions was studied. The results showed that all of
the studied isolates were resistant to Ap, Fc, Clin and Ct, while they were
sensitive to the antibiotic Ax, and the results were uneven with the rest of
the antibiotics. The yeasts showed absolute resistance to the heavy metal ions
NiSO4 and K2Cr2O7 and varied with
respect to the rest of the heavy metals. The ability of E. coli to resist
antibiotics and heavy metal ions has been studied. The results showed that the
two tested E. coli isolates were resistant to (Ap, Cf, Clin, Ct, Er, Fc,
Str, Rif, Nys, Pen, Ax, Itr, Tc and Cot), while the E. coli wasolate was
water. (2) The isolate from water is sensitive to the antibiotic Van, while the
isolate E. coli urine (1) isolated from the urine was sensitive to the
antibiotic Gen. While the two isolates were resistant to NiCl2,
NiSo4, CuSO4 and ZnSo4 and sensitive to CdCl2 and COCl2.
The results of isolation and transfer of plasmid DNA content of yeasts and
bacteria using agarose gel electrophoresis technique showed that isolate R.
mucilaginosa S6, isolate S. cervisiae S3, and bacteria E. coli
(1) contain plasmid DNA. The plasmid DNA content of the R. mucilaginosa S6
that the curing colonies did not bear the characteristic of hydrocarbon
hydrolysis, and this indicates that the genes responsible for this
characteristic are located on the plasmid DNA. The curing colonies showed
sensitivity towards the studied antibiotics and heavy metals with a percentage
of 1-92% except for NiSO4 and COCl2. Also, the results of the
neutralization of the isolate S. cervisiae S3 showed a loss of
resistance to antibiotics with a range of 3-93%. The results of neutralizing
the isolate E. coli showed a loss of antibiotic resistance with a range of
7-97%, except for Ax, which was confirmed by gel electrophoresis. The
conjugation process was successful between the donor isolate R. mucilaginosa
S6 and the neutral S. cervisiae S3 isolate as a receiver with a conjugation
frequency of 2.1 × 10-8. This study proved that the plasmid DNA transferred
from the isolate R. mucilaginosa S6 carries the genes for resistance to
the Er antibiotic and carries the genes responsible for the degradation of
hydrocarbon compounds. In addition, an attempt was made to conjugate across the
kingdoms between yeast R. mucilaginosa S6 as a host and E. coli
as a acceptor. The results showed that the plasmid DNA carrying Rif resistance
genes has the ability to move from yeast to bacteria with a conjugation
frequency of 5.8 × 10-8. In addition, the results showed that the neutralizing
bacteria have the ability to receive and localize the plasmid from the yeast
through the conjugation process, and the colonies resulting from the
conjugation showed the characteristic of hydrocarbon degradation, and this
confirms that the genes encoding the characteristic of hydrocarbon degradation
are located on the plasmid DNA.
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Pages:36-42
How to cite this article:
Sarah I Hayes, Badia A Malla Obaida "Testing the ability of plasmid DNA content in <i>Rhodotorula mucilaginosa</i> S6 to degradation hydrocarbon compounds and transfer through conjucation". International Journal of Biology Research, Vol 8, Issue 1, 2023, Pages 36-42
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