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Notes:
Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X Euro Biotechnology 2018
October 11-12, 2018
October 11-12, 2018 | Moscow, Russia
21st European
Biotechnology Congress
conferenceseries.com
Page 35
Single nucleotide polymorphisms and haplotype analyses in tilapia fish inferred from mtDNA
D-loop and Cyt-b regions
Ikpeme E V1, Udensi O U1, Ekerette E E 1 and Ozoje M O1,2
1University of Calabar, Nigeria
2Federal University of Agriculture Abeokuta, Nigeria
Objective: The research was aimed at analysing single nucleotide polymorphisms and haplotypes on D-loop and Cyt-b
regions of the mitochondrial DNA of tilapia fish.
Methods: Fifteen and thirteen tilapia fish were obtained from two populations, South-South (Domita farm)
and South West (Odeda farm). DNA extraction from fish tissue was done using Quick-gDNATM mini prep kit
after which PCR amplification was carried out. Sequencing of the two mtDNA regions were done using forward
primer 5’- GGATTYTAACCCYTRCCCC- 3’ and reverse 3’-AGTAAGTCAGGACCAAGCC-5’ for D-loop and
5’-GGATTTTAACCCTTACCCC-3’ and 3’-AGTAAAGTCAGGACCAAGCC-5’ for Cyt-b region. Statistical analyses
were carried out on the aligned sequenced data using MEGA version 6.06, DnaSP 5.1, Codon code aligner 6.06 as well
as NETWORK 4.6.1.1.
Results: mtDNA polymorphism was highest in the D-loop of South-South (SS) population with 176 polymorphic
sites, while South-West (SW) population had 162 polymorphic sites translating to 176, 162 and 144 SNPs with non-
synonymous substitutions higher than synonymous substitutions. Haplotype diversities (Hd) were 1.00±0.024 and
1.00±0.030 while nucleotide diversities were 0.168±0.086 and 0.161±0.084 for D-loop of SS and SW populations,
respectively. For Cyt b region, haplotype and nucleotide diversities were 0.91±0.003 and 0.051±0.016. Positive selection
was more on mtDNA D-loop of tilapia sampled from SS than those from the SW as well as Cyt-b region of tilapia fish
from SS. 28 haplotypes were identified among the tilapia from SS and SW with no shared haplotypes while 9 haplotypes
were identified from the Cyt-b region with haplotypes 4, 5, 6 and 7 shared between species. Median joining network
analysis revealed population-based clustering pattern. Demographic expansion was not significant using Tajima’s D and
Fu’s F statistics.
Conclusion: Higher SNPs were revealed in mtDNA D-loop when compared with mtDNA Cyt-b region of tilapia fish.
Ikpeme E V et al., J Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C5-100