مقايسه ژنتيكي ماهي سه خاره (Gasterosteus aculeatus) با استفاده از توالي يابي ژن سيتوكروم اكسيداز (COI) در سواحل جنوبي درياي خزر و شمالي درياي مديترانه

Genetic comparison of Three-spined stickleback (Gasterosteus aculeatus) by using sequencing of Cytochrome Oxidase 1 (COI) gene in the Southern Caspian Sea and the Northern Mediterranean Sea

ﻣﺎﻫﯽ ﺳﻪ ﺧﺎره )G. aculeatus( ﯾﮑﯽ از ﻣﻬﻤﺘﺮﯾﻦ ﮔﻮﻧﻪﻫﺎي ﻣﺪل اﺳﺖ ﮐﻪ ﺑﻪ ﺳﺎدﮔﯽ در ﺷﺮاﯾﻂ آزﻣﺎﯾﺸﮕﺎﻫﯽ ﭘﺮورش ﻣﯽﯾﺎﺑﺪ. ﺑﺎﺗﻮﺟﻪ ﺑﻪ ﻏﯿﺮﺑﻮﻣﯽ ﺑﻮدن اﯾﻦ ﮔﻮﻧﻪ در درﯾﺎي ﺧﺰر و ﻧﺎﻣﺸﺨﺺ ﺑﻮدن ﻣﻨﺸﺄ ورود آن و ﻫﻤﭽﻨﯿﻦ ﻣﺠﺰا ﺑﻮدن ﺟﻤﻌﯿﺖ ﻣﺎﻫﯿﺎن ﺳﻪ ﺧﺎره ﺟﻨﻮب ﻓﺮاﻧﺴﻪ در دﻟﺘﺎي رودﺧﺎﻧﻪ Rhone )ﺗﺎﻻب ﮐﺎﻣﺎرگ( از ﺳﺎﯾﺮ ﺟﻤﻌﯿﺖ-ﻫﺎي اروﭘﺎ و ﺑﻪ وﯾﮋه ﻣﺪﯾﺘﺮاﻧﻪ، در اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﻪ ﻣﻘﺎﯾﺴﻪ ژﻧﺘﯿﮑﯽ ﻣﺎﻫﯿﺎن ﺳﻪ ﺧﺎره ﺑﯿﻦ ﺧﻠﯿﺞ ﮔﺮﮔﺎن )ﺳﻮاﺣﻞ اﯾﺮان( ﺑﺎ ﺗﺎﻻب ﮐﺎﻣﺎرگ )ﺳﻮاﺣﻞ ﺟﻨﻮب ﻓﺮاﻧﺴﻪ( ﺑﺎ اﺳﺘﻔﺎده از ﺗﻮاﻟﯽ ﯾﺎﺑﯽ ژن COI ﭘﺮداﺧﺘﻪ ﺷﺪ. ﺑﺪﯾﻦ ﻣﻨﻈﻮر، ﺗﻌﺪاد 10 ﻗﻄﻌﻪ ﻣﺎﻫﯽ از ﻫﺮ ﻣﻨﻄﻘﻪ ﻧﻤﻮﻧﻪ ﺑﺮداري ﺷﺪ و ﺗﺎ زﻣﺎن اﻧﺠﺎم آزﻣﺎﯾﺶ در اﻟﮑﻞ اﺗﺎﻧﻮل 96% ﻧﮕﻬﺪاري ﺷﺪ. ﭘﺲ از اﺳﺘﺨﺮاج DNA و اﻧﺠﺎم واﮐﻨﺶ PCR ﺑﺎ اﺳﺘﻔﺎده از ﯾﮏ ﺟﻔﺖ آﻏﺎزﮔﺮ ﺳﯿﺘﻮﮐﺮوم اﮐﺴﯿﺪاز 1 )COI(، ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ از ﺗﻌﯿﯿﻦ ﺗﻮاﻟﯽ ﻧﺸﺎن داد ﮐﻪ ﻗﻄﻌﻪ ﺗﮑﺜﯿﺮ ﺷﺪه، 680 ﺟﻔﺖ ﺑﺎز ﻃﻮل دارد. ﺗﻌﺪاد اﻧﺪك ﻫﺎﭘﻠﻮﺗﺎﯾﭗﻫﺎ و ﺗﻨﻮع اﻧﺪك ﻫﺎﭘﻠﻮﺗﺎﯾﭗ-ﻫﺎ در ﻫﺮ دو ﻣﻨﻄﻘﻪ ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن دﻫﻨﺪه ﺗﻨﻮع ژﻧﺘﯿﮑﯽ ﭘﺎﯾﯿﻦ در آﻧﻬﺎ ﺑﻮد. ﻣﯿﺰان FST ﺑﻪ ﻋﻨﻮان ﺷﺎﺧﺺ ﺗﻤﺎﯾﺰ ژﻧﺘﯿﮑﯽ ﺟﻤﻌﯿﺖﻫﺎ 0/333 ﺑﻪ دﺳﺖ آﻣﺪ ﮐﻪ ﻧﺸﺎندﻫﻨﺪه ﺗﻤﺎﯾﺰ ﺑﺎﻻي دو ﺟﻤﻌﯿﺖ ﺑﻮد. ﺑﺎ وﺟﻮد ﻓﺎﺻﻠﻪ ژﻧﺘﯿﮑﯽ ﭘﺎﯾﯿﻦ )0/002( ﺑﯿﻦ ﻧﻤﻮﻧﻪﻫﺎي دو ﻣﻨﻄﻘﻪ ﺑﺮاﺳﺎس روش دو ﭘﺎراﻣﺘﺮ ﮐﯿﻤﻮرا )K2P(، ﺗﺮﺳﯿﻢ درﺧﺖ ﻓﯿﻠﻮژﻧﯽ ﻧﺸﺎن داد ﺟﻤﻌﯿﺖ ﻣﺎﻫﯿﺎن ﺳﻪ ﺧﺎره ﺧﻠﯿﺞ ﮔﺮﮔﺎن و ﺗﺎﻻب ﮐﺎﻣﺎرگ ﯾﮏ زﯾﺮ ﮐﻠﯿﺪ ﻣﺠﺰاﯾﯽ را ﺗﺸﮑﯿﻞ دادﻧﺪ. ﺣﺘﯽ ﮔﻤﺎن ﻣﯽرود اﯾﻦ دو ﺟﻤﻌﯿﺖ داراي ﻧﯿﺎي ﻣﺸﺘﺮك ﺑﺎﺷﻨﺪ، اﻣﺎ ﺑﻪ دﻟﯿﻞ درﺟﻪ ﺣﻤﺎﯾﺖ ﭘﺎﯾﯿﻦ ﺑﻮت اﺳﺘﺮپ(، رواﺑﻂ ﺑﯿﻦ ﮐﻠﯿﺪﻫﺎ ﺣﻞ ﻧﺸﺪ و ﻣﻨﺸﺄ ﺟﻤﻌﯿﺖ ﻣﺎﻫﯿﺎن ﺧﻠﯿﺞ ﮔﺮﮔﺎن ﻫﻢ ﺑﻪ درﺳﺘﯽ ﺷﻨﺎﺳﺎﯾﯽ ﻧﺸﺪ.

Three-spined stickleback (Gasterosteus aculeatus) is one the main model species which can be easily cultured in laboratory condition. Since it is invasive species in the Caspian Sea and the origin of its entrance is not clear. Furthermore, the three-spined population in South of France in Delta of Rhone River (Camargue wetland) is distinctive from other European populations especially Mediterranean population, therefore, Genetic comparison of three-spined stickleback between Gorgan bay (Iranian Coasts) and Camargue Wetland (South France Coasts) was studied by COI sequencing in this study. For this reason, 10 individual fish were sampled from each studied region and kept in Ethanol 96% until the time of experimental performance. After DNA extraction and PCR Reaction by one pair of Cytochrome Oxidase 1 (COI), Results of sequencing showed that the amplified sequence has 680 bp. The low number of haplotype and low haplotype diversity in both studied region dedicated the low genetic diversity of those populations. Level of FST as a Genetic distinction was 0.333 which is indicated high Genetic distinction of both population. Inspite of low Genetic distance (0.002) between two groups based on Kimura 2 Parameters method (K2P), phylogeny tree showed that the fish population of Gorgan bay and Camargue wetland formed a distinctive sub-clade. Even it is hypothesized that these two population have mutual origin but due to low supportive degree (bootstrap) the relationship between clades was not solved and the origin of three-spined sticklebacks of Gorgan bay was not properly evaluated.