fa ساختار ژنتیکی ماهی سفید (Rutilus frissi, Nordmann, 1840) در حوضه جنوبی دریای خزر (استان گیلان) با استفاده از توالی یابی DNA میتوکندریایی تخصصي Special پژوهشي Research <span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <a name="_Hlk95838394"><span b="" lang="FA" style="font-family:" zar="">به‌منظور بررسی امکان تمایز ژنتیکی و مقایسه ساختار ژنتیک جمعیت ماهی سفید با استفاده از روش‌های تعیین توالی </span></a><br> <span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">DNA</span></span> <span dir="LTR" new="" roman="" style="font-family:" times="">(</span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">DNA sequencing</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">)</span><span b="" lang="FA" style="font-family:" zar=""> ، تعداد ده عدد ماهی سفید از مناطق شرقی، مرکزی و غربی دریای خزر (سواحل استان گیلان) صید گردید. </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">DNA</span></span><span b="" lang="FA" style="font-family:" zar=""> 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 </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">ND1000</span></span><span b="" lang="FA" style="font-family:" zar="">) تعیین گردید. دو جفت پرایمر (</span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Forward</span></span><span b="" lang="FA" style="font-family:" zar=""> و </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Reverse</span></span><span b="" lang="FA" style="font-family:" zar="">) ژن‌ </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">16SrRNA</span></span><span b="" lang="FA" style="font-family:" zar=""> با استفاده از نرم افزار </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">GeneRuner</span></span><span b="" lang="FA" style="font-family:" zar=""> طراحی و سنتز شد. جهت تعیین توالی ژن </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">16SrRNA</span></span><span b="" lang="FA" style="font-family:" zar="">، پس از استخراج </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">DNA</span></span><span b="" lang="FA" style="font-family:" zar=""> و </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">PCR</span></span><span b="" lang="FA" style="font-family:" zar="">، محصول </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">PCR</span></span><span b="" lang="FA" style="font-family:" zar=""> بر روی ژل آگارز 1 درصد رانده شد و باندهایی در محدوده 500 </span><span arial="" lang="FA" style="font-family:">&ndash;</span><span b="" lang="FA" style="font-family:" zar=""> 600 جفت باز برای ژن </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">16SrRNA</span></span><span b="" lang="FA" style="font-family:" zar=""> تولید نمود. اختلاف تکاملی، درجه خویشاوندی و روابط فیلوژنی با استفاده از نرم افزار </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">MEGA11</span></span><span b="" lang="FA" style="font-family:" zar=""> به‌دست آمد. نتایج حاصله نشان داد که نمونه‌های ماهی سفید در سه منطقه صید شده دارای 95-97 درصد شباهت بوده و 5-3 درصد با یکدیگر اختلاف تکاملی دارند. درخت فیلوژنی نمونه‌های ماهی سفید با استفاده از روش‌ </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Neighbor</span></span><span dir="LTR" new="" roman="" style="font-family:" times="">-</span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Joining</span></span><span b="" lang="FA" style="font-family:" zar=""> ترسیم گردید و&nbsp; نشان داد که نمونه‌های مناطق غربی و شرقی هر کدام بر روی یک کلاستر مجزا قرار دارند که این می‌تواند نشانه تمایز جمعیت‌ها بر اساس ژن </span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">16SrRNA</span></span><span b="" lang="FA" style="font-family:" zar=""> باشد. با</span> <span b="" lang="FA" style="font-family:" zar="">توجه</span> <span b="" lang="FA" style="font-family:" zar="">به</span> <span b="" lang="FA" style="font-family:" zar="">تخریب</span> <span b="" lang="FA" style="font-family:" zar="">رودخانه‌های محل</span> <span b="" lang="FA" style="font-family:" zar="">مهاجرت،</span> <span b="" lang="FA" style="font-family:" zar="">آلودگی</span> <span b="" lang="FA" style="font-family:" zar="">دریای</span> <span b="" lang="FA" style="font-family:" zar="">خزر</span> <span b="" lang="FA" style="font-family:" zar="">و</span> <span b="" lang="FA" style="font-family:" zar="">عدم</span> <span b="" lang="FA" style="font-family:" zar="">وجود</span> <span b="" lang="FA" style="font-family:" zar="">موانع فیزیکی</span> <span b="" lang="FA" style="font-family:" zar="">در</span> <span b="" lang="FA" style="font-family:" zar="">این</span> <span b="" lang="FA" style="font-family:" zar="">پهنه</span> <span b="" lang="FA" style="font-family:" zar="">آبی،</span> <span b="" lang="FA" style="font-family:" zar="">ماهی</span> <span b="" lang="FA" style="font-family:" zar="">سفید</span> <span b="" lang="FA" style="font-family:" zar="">در</span> <span b="" lang="FA" style="font-family:" zar="">این</span> <span b="" lang="FA" style="font-family:" zar="">دو</span> <span b="" lang="FA" style="font-family:" zar="">منطقه هنوز</span> <span b="" lang="FA" style="font-family:" zar="">از</span> <span b="" lang="FA" style="font-family:" zar="">تنوع</span> <span b="" lang="FA" style="font-family:" zar="">ژنتیکی</span> <span b="" lang="FA" style="font-family:" zar="">برخوردار</span> <span b="" lang="FA" style="font-family:" zar="">می‌باشد.</span> <span b="" lang="FA" style="font-family:" zar="">بنابراین حفظ ساختار</span> <span b="" lang="FA" style="font-family:" zar="">ژنتیکی</span> <span b="" lang="FA" style="font-family:" zar="">جمعیت‌های</span> <span b="" lang="FA" style="font-family:" zar="">ماهی</span> <span b="" lang="FA" style="font-family:" zar="">سفید</span> <span b="" lang="FA" style="font-family:" zar="">مناطق مرکزی، شرقی و غربی در نوار ساحلی استان گیلان</span> <span b="" lang="FA" style="font-family:" zar="">دارای</span> <span b="" lang="FA" style="font-family:" zar="">اهمیت</span> <span b="" lang="FA" style="font-family:" zar="">اساسی می‌باشد.</span><b><span b="" lang="FA" style="font-family:" zar=""><span style="color:black"></span></span></b></span></span></span></span></span> <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Introduction:</span></b> <span lang="EN" new="" roman="" style="font-family:" times="">In order to investigate the possibility of genetic differentiation and to compare the genetic structure of Kutum population using DNA sequencing methods, ten Kutum</span><span new="" roman="" style="font-family:" times="">s</span><span lang="EN" new="" roman="" style="font-family:" times=""> from the eastern, middle and western regions of the Caspian Sea (coasts of Gilan province) were sampled from the mentioned areas. </span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Materials and Methods:</span></b><span new="" roman="" style="font-family:" times=""> DNA of three samples of each area and species was extracted using ammonium acetate method, their quantity and quality were determined using 1% agarose gel and 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, Kutum DNA were amplified by PCR. Then, PCR product was runned on 1% agarose gel and bands have been produced in the range of 500-600 bp. </span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span lang="EN" new="" roman="" style="font-family:" times="">Results and Discussion:</span></b><span lang="EN" new="" roman="" style="font-family:" times=""> Evolutionary difference (Maximum Likelihood), neighbor-joining and phylogeny relationships were obtained using MEGA11 software. The results show that the samples of Kutum in three areas have 97% similarity and 3- 5% evolutionary differences with each other. The phylogeny tree of Kutum samples was drawn using Neighbor-Joining methods. </span><span new="" roman="" style="font-family:" times="">In order to compare of phylogeny relationships of Kutum from three areas using Neighbor-Joining method, each western and eastern samples were placed in a separated cluster, which it could be a sign of population differences according to 16SrRNA gene. Due to the destruction of the rivers where they migrate, the pollution of the Caspian Sea and the lack of physical barriers in it, the kutum in these two regions still has genetic diversity.</span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span calibri="" style="font-family:"><b><span lang="EN" new="" roman="" style="font-family:" times="">Conclusion:</span></b> <span new="" roman="" style="font-family:" times="">So preserving the genetic structure of the kutum populations in the central, eastern and western regions on the coasts of Gilan province is important and essential.</span><span lang="EN" style="font-family:"Times New Roman",serif"></span></span></span></span> ساختار ژنتیکی, ماهی سفید, دریای خزر, DNA میتوکندری. Genetic structure, Kutum, Golden grey mullet, Caspian Sea, mtDNA. 34 48 http://aqudev.lahijan.iau.ir/browse.php?a_code=A-10-369-5&slc_lang=fa&sid=1 Mohammad Hassanzadeh Saber محمد حسن زاده صابر1 saber.merag@gmail.com 100319475328460011276 100319475328460011276 Yes 1-International Sturgeon Research Institute (ISRI), Iranian Fisheries Science Institute (IFSRI), Agricultural Research, Education and Extension Organization (AREEO), Rasht, Iran Omid Jafari امید جعفری1 jaafari.omid@yahoo.com 100319475328460011277 100319475328460011277 No Shirin Jamshidi شیرین جمشیدی1 jamshidi99@yahoo.com 100319475328460011278 100319475328460011278 No Mahmoud Mohseni محمود محسنی1 mahmoudmohseni73@gmail.com 100319475328460011279 100319475328460011279 No keywan Abbasi Ranjbar کیوان عباسی رنجبر keyvan_abbasi@yahoo.com 100319475328460011280 100319475328460011280 No Inland Waters Aquaculture Research Center, Iranian Fisheries Science Research Institute (IFSRI), Agriculture Research Education and Extension Organization (AREEO), Bandar-e Anzali, Iran. پژوهشکده آبزی پروری آبهای داخلی، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، بندر انزلی، ایران Siamak Bagheri سیامک باقری siamakbp@gmail.com 100319475328460011281 100319475328460011281 No Inland Waters Aquaculture Research Center, Iranian Fisheries Science Research Institute (IFSRI), Agriculture Research Education and Extension Organization (AREEO), Bandar-e Anzali, Iran. پژوهشکده آبزی پروری آبهای داخلی، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، بندر انزلی، ایران Esmail Abdolahzadeh اسماعیل عبداله زاده1 abdollahzadeh@rocketmail.com 100319475328460011282 100319475328460011282 No Maryam Forouzad مریم فروزد zhm.forouzad@gmail.com 100319475328460011283 100319475328460011283 No Iranian Fisheries Science Research Institute (IFSRI), Agriculture Research Education and Extension Organization (AREEO), Tehran, Iran. موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج جهاد کشاورزی، رشت، ایران