Detection of Genetic Relationship and Genetic Variation of the Four Species of Real Flies Using Molecular Markers
Objective: To uncover the genetic relationship and the genetic variation of four types of true flies, which are as follows: Musca domestica, Chrisomya megacephala, Sarcophaga africa Tabanus regulars, using RAPD-PCR markers.
Methods of work: Samples were collected from Tarmiyah district in Baghdad governorate using traps. The species of flies were diagnosed at the Research Center and Museum of Natural History at the University of Baghdad. The species were preserved using 70% ethanol alcohol, the DNA extraction process was carried out, the purity and concentration of the DNA was measured and adjusted using a nano drop device, then the RAPD- PCR reactions were performed using (17) primers and the results were migrated to the acarose gel and the packages were photographed using a digital camera, after which the statistical analysis was performed. Her and the results are saved.
Results: The results of the RAPD markers showed different types of bands, and the total loci identified by the primers on the samples genome were (151) loci, of which (11) are general loci and (140) are differentiated loci. A locus, while the two primers produced (P5, P7), the lowest number of loci reached (6) loci. The total number of produced bands was (276) (a band of which (231) polymorphic bands and (44) monomorphic bands. as for the general variation ratio of the produced primers, it was 84%. The study showed the presence of distinctive bands, the total of the distinctive bands was (85), and of which (67) were unique bands and (18) were absent. (12) bands As for the absent bands, the house fly was distinguished by the lowest number of absent bands (3) band, while the horse fly got the highest number of absent band, reaching (6) bands. From the results of the interactions of the genetic distance, the values of the genetic distance ranged between (0.652 - 0.975), where the least genetic distance was between the meat fly and the horse fly, as it reached (0.652). This indicates that these two types are the most similar among the studied species, and it was the highest genetic distance between It is between the metal fly and the horse fly with a genetic distance (0.975), which indicates that they are more different within the studied species.
Conclusion: There is a close degree of affinity between the four types of flies, even though they belong to families, so molecular markers can be used in classifying insects, especially flies, accurately and determining their genetic fingerprint instead of adopting the phenotypic features in the classification because molecular markers are fixed traits and do not change with changing conditions.
Al-Sugmiany, Rafea Zaidan (2017). The Use of Morphological and Molecular Markers to Assess the Genetic Performance for a number of genotypes of (vicia faba.L) and their di alleles crossing. College of Education for Pure Sciences. Tikrit University.
Bajpai N. and Tewari R.R.(2010). Genetic characterization of sarcophagid flies by Random Amplified Polymorphic DNA- Polymerase Chain Reaction (RAPD-PCR), (Hamilton, 1822) using RAPD markers and implications for its Conservation, J. Cell Mol. Biol., 8(2), 77-85.
Benecke M, Wells JD, (2001). DNA techniques for forensic entomology. In Forensic Entomology: Utility of Arthropods in Legal Investigations. Byrd. Boca Raton: CRC Press; 341-352
Boyce TM, ME Zwike and Aquadro CF. (1989). Mitochondrial DNA in the bark weevils: size, structure and heteroplasmy. Genetics 123: 825-836.
Brito L.G., Regitano L.C.A., Huacca M.E.F., Carrilho E., Paes M.J. and Borja G.E. (2008). Genotype characterization of the Haematobia irritans (Diptera : Muscidae) from Brazil, Dominican Republic and Colombia based on randomly amplified polymorphic DNA (RAPD) analysis, Revista Brasileira de Parasitologia Veterinaria, 17(4), 179-184
CORDEIRO GLÁUCIA , DA CUNHA MARINA S, MARINA S CAROLINA R, JORGE ISAAC R, DERGAM JORGE A, FERREIRA PAULO S.F. (2019). Molecular identification of three species of Oncideres (Coleoptera : Cerambycida) using RAPD markers . An. Acad.bras.Cienc vol.91 no.3 Rio de janeiro.
Infante-Malachias, M.E.V.; Yotoko, K.S. and Azeredo-Espin, A.M.L.(1999). Random Amplified Polymorphic DNA of screwworm fly Populations (Diptera: Calliphoridae), from South-east of Brazi l and North of Argentina. Genome 42: 772-779.
Malviya S, Tewari RR, Agarwal UR.(2015). Genetic Relationship between the Muscids Using RAPD-PCR as marker. International Research Journal of Biological Sciences; 4(1):66-70.
Malviya S., Bajpai N. and Tewari R.R. (2011). Genetic Relatedness among three populations of housefly, Musca domestica L. using RAPD - PCR marker, Int. J. Pharm. BioSci., 2(4), 198-204.
Malviya S., Bajpai N. and Tewari R.R.(2012). RAPD-PCR Based genetic relationship of muscid flies (Diptera: Muscidae), Int. J. Pharma and BioSci., 3(3), 1018- 1024).)
MONDINI L., NOORANI A. and PAGNOTTA M.A. (2009). Assessing plant genetic diversity by molecular tools, Diversity 1: 19 -35.
Nei, M. and W.H. Li (1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceeding of the National Academy of Science, USA. 74, 5269-5273. C. F. by Henry, R.J.
Sambrook, J., Fritch, E. F. and Maniatis, J. (1989). Molecular cloning, a laboratory Manual. 2nd edition. Cold spring Harbor laboratory press, New York.
Sharma M, Singh D, Sharma AK.(2015). Mitochondrial DNA based identification of forensically important Indian flesh flies Diptera: Sarcophagidae. Forensic Science International, 247:1-6.
Schuman, H. (1992). Systimatiche Gliederung der ordung Diptera mitbesnder Beruksichtinum derin deatsc land, Vorkommenden Familien, Dtsch. Ent. Z. N. E., 39: 103-116.
SINGH VK, JOSHI PC, BISHT SPS, KUMAR S, NATH P, AWASTHI S and MANSOTRA DK. (2016). Molecular characterization of butterflies and its significances in taxonomy. J Entomol Zool Stud 4(2): 545-547
Sultan, Ammar Ahmed , Qadir ,Hind Tahir.(2015). Studying the genetic relationship among three populations for Musca domestica L. (Diptera : Muscidae) in Iraq by using RAPDPCR Technique . Advances in Life Science and Technology www.iiste.org ISSN 2224-7181 (Paper) ISSN 2225-062X (Online) Vol.35.
SUNITHA V, SINGH TVK, RAMESH BABU V and SATYANARAYANA J. (2015). Genetic diversity assessment using RAPD primers in insecticide resistant populations of diamondback moth Plutella xylostella (Linn.). J Appl Nat Sci 7(1): 219-225.
Williams JGK, Hanafey MK, Rafalski JA, Tingey SV.(1995). Genetic analysis using random amplified polymorphic DNA markers. In: Wu R, editor. Recombinant DNA methodology II. Academic Press;. pp. 849–883
Williams, J.G.K; A.R., KubelikK.J., , Livak, J.A. Rafalski, and S.V. Tingey, (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Rese. 18: 6531-6535.
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