A study on genetic differentiation in two species of Iranian bleaks, (Alburnus mossulensis) and (Alburnus caeruleus) (Teleostei, Cyprinidae) using simple sequence repeats


1 1. Faculty of Marine Science, Tarbiat Modares University, P.O.Box:46414-356, Noor, Iran. 2. Faculty of Natural Resources, Isfahan University of Technology, Isfahan, 84156-83111, Iran. * Corresponding author?s E-mail: kalbassi_m@modares.ac.ir

2 0


The genetic structure of the genus Alburnus is not well known and the phylogenetic relationships among its species are uncertain. In the present study, simple sequence repeats (SSRs or microsatellites) were used to evaluate genetic diversity and genetic differentiation between Alburnus mossulensis Heckel, 1843 from Kashgan River in Lorestan province and Alburnus caeruleus Heckel, 1843 from Gamasiab River in Kermanshah province. Thirty specimens from each species were collected and their genomic DNA was extracted. Polymerase chain reaction was performed using four pairs of SSR markers, including CypG24, BL1-2b, BL1-98 and Rser10, from which a total of 480 bands were amplified. The average observed and expected heterozygosities for both species were similar. In both species, except for Rser10 locus, all loci deviated from the Hardy-Weinberg equilibrium (P < 0.05). Average genetic distance and Fst values between the two species were 0.361 and 0.04, respectively. The AMOVA analysis revealed more interspecific (94%) than intraspecific (4%) genetic variation. Although four sets of SSR markers developed for other cyprinids showed high level of polymorphisms in the Iranian bleaks, they showed low genetic differentiation between them. Study on the possibility of genetic differentiation of two examined species by more microsatellite loci or other molecular markers such as amplified fragment length polymorphisms (AFLP) are recommended.


Balloux, F. and Lugon-Moulin, N. (2002) The estimation of population differentiation with microsatellite markers. Molecular Ecology, 11: 155-165. Beaumont, R.A. & Hoare, K. (2003) Biotechnology and Genetics in Fisheries and Aquaculture. Blackwell Publishing, Oxford. P. 176. Belfiore, N.M. & Anderson, S.L. (2001) Effects of contaminants on genetic patterns in aquatic organisms: a review. Mutation Research, 489: 97-122. Birgitte, J. Hansen, M. & Loeschcke, V. (2005) Microsatellite DNA analysis of northern pike (Esox lucius) population: Insights into the genetic structure and demographic history of a genetically depauperate species. Biological Journal of Linnaean Society, 84: 1-11. Brookfield, J.F.Y. (1996) A simple new method for estimating null allele frequency from heterozygote deficiency. Molecular Ecology, 5: 453-455. Castric, V. Bernatchez, L. Belkhir, K. & Bonhomme, F. (2002) Heterozygotedeficiencies in small lacustrine populations of brook charr Salvelinus fontinalis Mitchill (Pisces, Salmonidae): a test of alternative hypotheses. Heredity, 89: 27-35.Coad, B.W. (2009) Alburnus zagrosensis, a new species of fish from the Zagros Mountains of Iran (Actinopterygii: Cyprinidae). Zoology in the Middle East, 48: 63-70. Coad, B. (2013) Freshwater fishes of Iran. [Online]. Available at: http://www. briancoad. com/Species%20Accounts/Contents%20new.htm[Accessed 6 May 2013]. Dubut, V. Martin, J.F. Costedoat, C. Chappaz, R. & Gilles, A. (2009) Isolation and characterization of polymorphic microsatellite loci in the freshwater fishes Telestes souffia and Telestes muticellus (Teleostei: Cyprinidae), Molecular Ecology Research, 9: 1001-1005. Dubut, V. Sinama, M. Martin, J. Meglécz, E. Fernandez, J. Chappaz, R. Gilles, A. & Costedoat, C. (2010) Cross-species amplification of 41 microsatellites in European cyprinids: A tool for evolutionary, population genetics and hybridization studies. BMC Research Notes, 3: 135. Epperson, B.K. (2003) Geographical Genetics. Monographs in Population Biology.Princeton University Press, Princeton. Fallah Bagheri, F. Dorafshan, S. Pourkazemi, M. Keivany, Y. & Chakmedouz Qasemi F. (2013) Genetic analysis of wild common carp, Cyprinus carpio L. in the Anzali wetland, the Caspian Sea. Iranian Journal of Fisheries Sciences, 12: 1-11. Freeland, J.R. (2007) Molecular Ecology. Wiley Publication, London, P. 400. Keivany, Y. Nasri, M. Abbasi, K. & Abdoli, A. 2015. Atlas of Inland Water Fishes of Iran. Iran Department of Environment. Liu, K. & Muse, V. (2004) PowerMarker: Integrated analysis environment for genetic marker data. Bioinformatics, 21: 2128-2129. Liu, Z. (2007) Aquaculture genome technologies. Blackwell Publishing. New York, P. 551. Liu, Z.J. & Cordes, F.J. (2004) DNA marker technologies and their applications in aquaculture genetics. Aquaculture, 238: 1-37. Matter, E.E., Elserafy, S.S., Zowail, M.E.M. & Awwad, M.H. (1992) Genotoxic effect of carbamyl insecticide (sevin) on the grass carp Ctenopharyngodon idella. Egyptian Journal of Histology, 15: 9-17. Nei, M., Tajima, F. & Tateno, Y. (1983) Accuracy of estimated phylogenetic trees from molecular data. Journal of Molecular Evolution, 19: 153-170. Okumus, I. & Ciftci, Y. (2003) Fish population genetics and molecular markers: II- molecular markers and their applications in fisheries and aquaculture. Turkish Journal of Fisheries and Aquatic Sciences, 3: 51-79. Pourkazemi, M. & Razikazemi, S. (2011) Failure of PCR-RAPD technique to differentiate sex in Mahisefied (Rutilus frisii kutum) from the South Caspian Sea. Dorafshan et al., 203Caspian Journal of Environmental Sciences, 9: 235-242. Pourkazemi, M. Khoshkholgh, M.R. Nazari, & S. Azizzadeh Pormehr, L. (2012) Genetic relationships among collections of the Persian sturgeon, Acipenser persicus, in the south Caspian Sea detected by mitochondrial DNA Restriction fragment length polymorphisms. Caspian Journal of Environmental Sciences, 10: 215-226. Raymond, M. & Rousset, F. (1995) GENEPOP, version1.2: population genetics software for exact tests and ecumenicist. Journal of Heredity, 86: 248-249. Rice, W.R. (1989) Analyzing tables of statistical tests. Evolution, 43, 223-225. Rousset, F. (2004) Genetic Structure and Selection in Subdivided Populations. Princeton: Princeton University Press. P. 288. Sambrook, J. Fritsch, E.F. & Maniatis, T. (1989) Electrophoresis of RNA through Gels Containing Formaldehyde: Molecular Cloning. 2nd ed. Cold Spring Harbour, CSH Laboratory Press, New York. Schneider, S. Roessli, D. & Excoffier, L. (2000) Arlequin: a Software for Population Genetics Data Analysis, Version 2.000. Genetics and Biometry Laboratory, Department of Anthropology, University of Geneva, Switzerland. Shirangi, S.A. Kalbassi, M.R. & Dorafshan, S. (2011) Microsatellite polymorphism reveals low genetic differentiation between fall and spring migratory forms of endangered Caspian trout, Salmo trutta caspius (Kessler, 1870). Caspian Journal of Environmental Sciences, 9: 9-16. Wang, Z. Zhou, J. Ye, Y. Wei, Q. & Wu, Q. (2006) Genetic structure and low-genetic diversity suggesting the necessity for conservation of the Chinese longsnout catfish, Leiocassislongirostris (Pisces: Bagriidae). Environmental Biology of Fishes, 75: 455-463. Wright, B.S. (1951) The genetical structure of populations. Animal Genetics, 15: 323-354. Zhang, X.G. & Tong, J.G. Xiong, B.X. (2006) Applications of microsatellite markers in studies of genetics and breeding of fish. Chin. Journal of Agriculture Biotechnology, 3: 83-87. Zhao, N.A. Shao, W. Zhu, Z. Brosse, B. & Chang, J. (2005) Microsatellite assessment of Chinese sturgeon (Acipenser sinensis). Journal of Applied Ichthyology, 21: 7-13.