Genetic relationships among collections of the Persian sturgeon, Acipenser percicus, in the south Caspian Sea detected by mitochondrial DNA Restriction fragment length polymorphisms


1 M. Khoshkholgh1*, M. Pourkazemi2, S. Nazari1, L. Azizzadeh Pormehr2

2 O. Rafieyan*1, A. A. Darvishsefat2, S. Babaii1, A. Mataji1


In the present study, mitochondrial DNA polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to assess the population structure and genetic relationships among six Persian sturgeon, Acipenser persicus populations from south Caspian Sea along the Iranian coast. The complete nucleotide dehydrogenase subunit 5 (NADH 5) region of mtDNA amplified by PCR was digested with five restriction enzymes. In total, 154 individuals from six populations including: Guilan (Zone1-2), Mazandaran (Zone 3 and 5), Golestan (Zone 4) and Sefidroud River, from south Caspian Sea along the Iranian coast were analyzed using five restriction endonucleases (Rsa ?, Hinf ?, HaeIII, Mbo ? and Cfr13?), yielding 17 haplotypes. Samples from Sefidroud River were clearly identified by cluster and molecular variance model (AMOVA) analyses. This collection showed dominant haplotypes that were little in populations from the other geographic areas. The mean haplotype diversity (h) and nucleotide diversity (?) were 0.739?0.038 and 0.0105?0.0043, respectively. Based on heterogeneity test haplotype frequencies of Persian sturgeon populations and Monte-Carlo with 1000 replicates in PCR-RFLP method significant differences were seen (?2 =37.12, P< 0.0001) and these results showed that haplotype distribution in different location were significant and populations of Sefidroud were statistically significant (P< 0.0001). This result suggests that the unique genetic structure of Sefidroud River represents a highly valuable genetic resource and should now be treated as demographically independent and managed separately.


Ataei, M., Pourkazemi, M., Hasanzadeh Kiabi, B and Shedaei, M. (2004). Analysis of mtDNA control region polymorphism of Persian sturgeon, Acipenser persicus, in Sefidrud River and Southeast region of Caspian Sea. Journal of Environment of Shahid Beheshti University 2(1): 27-31.(In Persian)


Avise J.C. (1994). Molecular Markers, Natural History, and Evolution. Chapman and Hall, New York, NY. 511 pp. Bemis WE, Kynard B (1997) Sturgeon rivers: an introduction to acipenseriform biogeography and life history. Environmental Biology of Fishes, 48:167–183



Billington N. and Hebert D.N. (1991). Mitochondrial DNA diversity in fishes and its implications for introductions. Canadian Journal of Fisheries and Aquatic Sciences 48 (Supplement 1), 80–94.


Billington, N. (2003). Mitochondrial DNA. Pages 59–100 in E. M. Hallerman, editor. Population genetics: principles and applications for fisheries scientists. American Fisheries Society, Bethesda, Maryland.


Birstein V.I, Bemis W.E, and Waldman J.R. (1997). The threatened status of Acipenseriform species: a summary. Environmental Biology of Fishes, 48, 427-435.


Brown K.H. (2008). Fish mitochondrial genomics: sequence, inheritance and functional variation. Journal of Fish Biology 72, 355–374.


Caughley G. & Gunn A. (1996) Conservation Biology in Theory and Practice. Blackwell Science, Cambridge, Massachusetts


Excoffier L., Smouse P.E., and Quattro J.M. (1992). Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics 131, 479-49l.


Excoffier L., Laval G., and Schneider S. (2005). Arlequin Ver. 3.0: an integrated software package for population genetics data analysis. Evolution Bioinformatics, 1, 47–50.

Genetic relationships of the Persian sturgeon224


Felsenstein, J., (1985). Confidence Limits on Phylogenies: An Approach Using Bootstrap, Evolution, 1985, vol. 39,pp. 783–791.


Felsenstein J. (1995). PHYLIP, Phylogeny inference package, version 3.57c. Department of Genetics, SK-50, University of Washington, Seattle. Frankham, R., Ballou, J. D. and Briscoe, D. A. (2002). Introduction to Conservation Genetics. Cambridge University Press, Cambridge, UK.


Hillis, D and Moritz, M. C. (1990). Molecular taxonomic. Sinauer associate, Inc. Publishers. Massachusetts. Ivanov, V. P., Vlasenko, A. D., Khodorevskaya, R. P. & Raspopov, V. M. (1999).Contemporary status of Caspian sturgeon (Acipenseridae) stock and its conservation. Journal of Applied Ichthyology 15,103-105.


Khoshkholgh, M.(2007). Comparision on genetic structure of Persian sturgeon (Acipenser persicus) in the Southern coast and Russian sturgeon (A. gueldenstaedtii) in the South and North Caspian Sea using Microsatellite techniques. Ph. D. Thesis. Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran.126pp


Lande, R. (1988). Genetics and demography in biological conservation. Science, 241:1455-1460.


Ludwig A, Kirschbaum F (1998) Comparison of mitochondrial DNA sequences between the European and the Adriatic sturgeon. Journal of Fish Biology 52:1289–1291


Ludwig, A. (2006). A sturgeon view on conservation genetics. European Journal of Wildlife Resources 52(1), 3-8.


McElroy, D.M., Moran, P., Bermingham, E., and Kornfield, I., (1992). REAP: An Integrated Environment for the Manipulation and Phylogenetic Analysis of Restriction Data, Journal of Heredity, 1992, vol. 83, pp. 153–158.


McNeely J.A., Miller K.R., Reid W.V., Mittermeier R.A. & Werner T.B. (1990) Conserving the World’s Biological Diversity. IUCN, World Resources Institute, Conservation International, WWF-US and the World Bank, Washington, DC.


Moghim M., Kor D., Tavakolieshkalak M. & Khoshghalb M.B. (2006). Stock status of Persian sturgeon (Acipenser persicus,Borodin 1897) along the Iranian coast of the Caspian Sea.Journal of Applied Ichthyology22, 99-107.


Moritz C. (1994). Applications of mitochondrial DNA analysis in conservation: a critical review. Molecular Ecology 3:401–411 Nei, M.,(1972). Genetic Distance between Populations, Am.Naturalist, 1972, vol. 106, pp. 283–292.


Nei, M., and Tajima, F., (1981). DNA polymorphism detectable by restriction endonucleases. Genetics 97: 145-163.


Lalouie, F., (1996) Study on Nature Reproduction in sturgeon of the south Caspian Sea in the Tajan and Gorgan Rivers. Iranian Journal of Fisheries Sciences , 5(4), 17-30. (In Persian).


Pikitch E.K., Doukakis P., Lauck L., Chakrabarty P. & Erickson D.L. (2005). Status, trends and management of sturgeon and paddlefish Fish and Fisheries 6, 233–265.


Pourkazemi M. (1996). Molecular and biochemical genetic analysis of sturgeon stocks from the South Caspian Sea. Ph.D. thesis.University of Wales, Swansea.


Pourkazemi M., Skibinski, D.O.F. & Beardmore J.A., (1999). Application of mtDNA d-loop region for the study of Russian sturgeon population structure from Iranian coastline of the Caspian Sea. Journal of Applied Ichthyology15, 23–28.


Pourkazemi, M. (2006). Caspian Sea sturgeon Conservation and Fisheries: Past present and Future. Journal of Applied Ichthyology 22 (Supplement 1). 12-16.


 Pourkazemi, M. (2008). Comprehensive project of genetic stock assessment in sturgeon species of the Caspian Sea. Iranian Fisheries Research Institute (Final Project Report). International Sturgeon Research Institute. 354pp.(In Persian)


Putilina L.A. (1985). Quantitative and qualitative assessment of spawning population of Persian sturgeon in the Pourkazemi et al.,225Volga river. "Rybnoe khozyaistvo (Fishing industry)". 10: 71-72.


 Putilina L.A. and E.N. Artyukhin. (1985). Some features of Persian sturgeon migration in the lower Volga River. Ibid. P. 32-34.Qasemi A., Pourkazemi M. & Kalbasi M. (2004). Genetic variation in ship sturgeon (Acipenser nudiventris) from the south Caspian Sea using PCR-RELP. Iranian Scientific Fisheries Journal 4, 151-163.(In Persian)


Rezvani Gilkolaei S. (1997). Molecular population genetic studies of sturgeon species in the South Caspian Sea. Ph.D. thesis. University of Wales, Swansea.


Rezvani Gilkolaie S. (2000). Study of mtDNA variation of Russian sturgeon population from the south Caspian Sea using RFPL analysis of PCR amplified ND5/6 gene regins. Iranian Journal of Fisheries Sciences2, 87-98.


Roff, D. and Bentzen, P., (1989). The Statistical Analysis of Mitochondrial DNA Polymorphisms: Chi-Square and the Problem of Small Samples, Molecular Biology and Evolution1989,vol. 5, pp. 539–545.


Schneider, S., D. Roessli, and L. Excoffier. (2000). Arlequin: a software for population genetics data analysis, version 2.0. Genetics and Biometry Laboratory, Department of Anthropology, University of Geneva, Geneva, Switzerland.


Shabani A., Pourkazemi M., Kamali A., Rezvani Gilkolaie S. & Vitskayal V.V. (2003). Study of Genetic variation in stellate sturgeon (Acipenser stellatus) from north (Volga river) and south (Gorganrud river) Caspian Sea using PCR-RFLP of mitochondrial. gene. Iranian journal of Marin Science and Technology 4, 159-169.(In Persian)


Slatkin, M. (1985). Rare alleles as indicators of gene flow. Evolution 39, 53–65.


Swofford, D.L., (2002). PAUP: Phylogenetic Analysis Using Parsimony (and Other Methods): Version, 4.0b10,


Sunderland:Sinauer. Tavakoli, M., Parafkandeh Haghighi, F., Khoshghalb M.B., Keymaram, F & Taghavi, A. (2010). Stock status of Persian sturgeon (Acipenser persicus,Borodin 1897) along the Iranian coast of the Caspian Sea.Iranian Scientific Fisheries Journal 1, 1-8.(In Persian) Vecsei


 P. & Artyukhin E. (2001). Threatened fishes of the world: Acipenser persicus Borodin, 1897 (Acipenseridae) Environmental Biology of Fishes 61, 160.


Weir B.S.& Cockerham C.C. (1984). Estimating F-statistics for the analysis of population structure. Evolution 38, 1358–1370.