Journal of Aquaculture Development

fa ساختار ژنتیکی کفال طلایی (Chelon auratus, Risso 1810) با استفاده از توالی‌یابی 16SrRNA-mtDNA در حوضه جنوب غربی دریای خزر (سواحل استان گیلان) Genetic structure of golden mullet (Chelon auratus, Risso 1810) stocks by 16SrRNA-mtDNA sequencing on the southwestern basin of the Caspian Sea (Guilan province) تخصصي Special پژوهشي Research <div style="text-align: justify;">     به‌منظور بررسی امکان تمایز ژنتیکی و مقایسه ساختار ژنتیک جمعیت کفال طلایی با استفاده از روش‌های تعیین توالی DNA  (DNA sequencing)، تعداد 9 عدد کفال طلایی از مناطق شرقی، مرکزی و غربی دریای خزر (سواحل استان گیلان) با استفاده از تور محاصره‌ای (چشمه ریز پره) در خرداد 1400 صید گردید. DNA سه نمونه ماهیان از هر منطقه با استفاده از روش استات آمونیوم استخراج شد و کمیت و کیفیت آن‌ها با استفاده از ژل آگارز 1 درصد و دستگاه نانودراپ (مدل ND1000) تعیین گردید. دو جفت پرایمر (Forward و Reverse) ژن‌ 16SrRNA با استفاده از نرم افزار GeneRuner طراحی و سنتز شد. جهت تعیین توالی ژن 16SrRNA، پس از استخراج DNA و PCR، محصول PCR بر روی ژل آگارز 1 درصد رانده شد و باندهایی در محدوده 500 – 600 جفت باز برای ژن 16SrRNA تولید نمود. اختلاف تکاملی، درجه خویشاوندی و روابط فیلوژنی با استفاده از نرم افزار MEGA11 به‌دست آمد. نتایج حاصله نشان داد که نمونه‌های کفال طلایی در مناطق صید شده دارای 96 درصد شباهت بوده و 4 درصد با یکدیگر اختلاف تکاملی دارند. درخت فیلوژنی نمونه‌های کفال با استفاده از روش‌های Maximum parsimony، Maximum Likelihood و Neighbor-Joining ترسیم گردید. در مقایسه بین سه روش بررسی روابط فیلوژنی کفال طلایی از سه منطقه نمونه‌برداری شده، در هر سه حالت (Maximum Parsimony, Maximum Likelihood, Neighbor-Joining)، نمونه‌های مناطق غربی و مرکزی بصورت مشترک در یک کلاستر و روی یک شاخه قرار دارند که نشان‌دهنده تمایز اندک این دو جمعیت بر اساس ژن 16SrRNA می‌باشد.</div> <div style="text-align: justify;">Introduction: The golden mullet (Chelon auratus) and kutum (Rutilus frisii) are among the valuable and economic species in the southern basin of the Caspian Sea bony fish stocks, accounting for over 85% of the total catch of these fish. A review of the reproduction and catch statistics of kutum and mullet over various years shows these stocks result from artificial reproduction and mass release of juveniles, and the share of natural reproduction in the restoration of stocks has decreased and reached its lowest point. This trend has side effects and will gradually cause the destruction of genetic stocks and the gradual destruction of the gene bank, which will cause a decrease in growth rate, a decrease in average length, a decrease in the percentage of spawning, and an increase in deformed larvae over 25 to 40 years. Therefore, knowledge of the genetic structure of these fish, obtained by examining population estimates and obtaining the level of genetic diversity and inbreeding and the adverse effects resulting from it, seems essential for the conservation and sustainable production of this economic species. Material and Methods: In order to investigate the possibility of genetic differentiation and to compare the genetic structure of mullet population using DNA sequencing methods, 9 mullets from the eastern, middle and western regions of the Caspian Sea (coasts of Guilan province) using a purse seine in June 2021 were sampled from the mentioned areas. DNA from three samples of each area was extracted using the ammonium acetate method; their quantity and quality were determined using 1% agarose gel and a Nanodrop device (ND1000), respectively. One pair of primers (Forward and Reverse) of 16SrRNA gene were designed and synthesized using GeneRuner software. In order to determine the sequence of the 16SrRNA gene, mullet DNA were amplified by PCR. Then, the PCR product was run on a 1% agarose gel, and bands were produced between 500 and 600 bp. Evolutionary difference (maximum likelihood), neighbor-joining and phylogeny relationships were obtained using MEGA11 software. Results and Discussion: The results show the mullet samples in the three areas have 96% similarity and 4% evolutionary differences with each other. The phylogeny tree of mullet samples was drawn using Maximum parsimony, maximum Likelihood and neighbor-joining methods. In order to compare among three methods in the golden mullet from the three areas, the samples of western and central areas are located together in one cluster and on the same branch, which indicates the slight distinction of these two populations according to the 16S rRNA gene. In the present study on golden mullet, the short geographical distance of sampling stations in the coastal waters of the Caspian Sea in Guilan Province and the absence of physical barriers such as heat, salinity, etc. (Rezvani et al., 2012) confirm the existence of gene flow in the three sampled areas, indicating little genetic differentiation in this species. The results obtained indicate that the inappropriate method of reproduction and release of larvae can be the main reason for the lack of differentiation, as well as the cause of the occurrence of a genetic bottleneck among the studied populations of golden mullet. Conclusion: Providing the possibility of natural reproduction of the broodstock is the best way to prevent the continuation of this process. Considering the need for artificial reproduction, it is necessary to consider the separation of populations during artificial reproduction and release.</div> تمایز ژنتیکی, جمعیت کفال طلایی, جنوب غربی دریای خزر, 16SrRNA Genetic distinct, Golden mullet population, Sothern west of the Caspian Sea, 16SrRNA 55 70 http://aqudev.lahijan.iau.ir/browse.php?a_code=A-10-369-6&slc_lang=fa&sid=1 Mohammad Hassanzadeh Saber1 محمد حسن زاده صابر1 saber.merag@gmail.com 100319475328460012214 100319475328460012214 Yes 1-International Sturgeon Research Institute, Iranian Fisheries Science Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rasht, Iran 1-انستیتو تحقیقات بین‌المللی ماهیان خاویاری، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، رشت، ایران AliNaghi Sarpanah1 علی نقی سرپناه1 sarpanah5050@gmail.com 100319475328460012215 100319475328460012215 No Omid Jafari1 امید جعفری1 jaafari.omid@yahoo.com 100319475328460012216 100319475328460012216 No Shirin Jamshidi1 شیرین جمشیدی1 jamshidi99@yahoo.com 100319475328460012217 100319475328460012217 No Mahmoud Mohseni1 محمود محسنی1 mahmoudmohseni73@gmail.com 100319475328460012218 100319475328460012218 No Esmail Abdolahzadeh1 اسماعیل عبداله زاده1 abdollahzadeh@rocketmail.com 100319475328460012219 100319475328460012219 No Keyvan Abbasi کیوان عباسی keyvan_abbasi@yahoo.com 100319475328460012220 100319475328460012220 No پژوهشکده آبزی‌پروری گیلان، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، بندر انزلی، ایران Siamak Bagheri2 سیامک باقری2 siamakbp@gmail.com 100319475328460012221 100319475328460012221 No Maryam Forouzad مریم فروزد zhm.forouzad@gmail.com 100319475328460012222 100319475328460012222 No موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، تهران، ایران Mohammad Sayyad Bourani2 محمد صیاد بورانی2 mohammadborani@yahoo.com 100319475328460012223 100319475328460012223 No