بررسی تنوع ژنتیکی لاکتوباسیلوس‌های جدا شده از نمونه‌های شیر گاو به روش RAPD-PCR
محورهای موضوعی : فصلنامه کیفیت و ماندگاری تولیدات کشاورزی و مواد غذاییحکیمه قلعه خانی 1 , معصومه حاجی رضایی 2 , آرتادخت توکلی 3
1 - کارشناسی ارشد، گروه میکروبیولوژی، واحد کرمان، دانشگاه آزاد اسلامی، کرمان، ایران
2 - استادیار، گروه میکروبیولوژی، واحد کرمان، دانشگاه آزاد اسلامی، کرمان، ایران
3 - استادیار، گروه میکروبیولوژی، واحد کرمان، دانشگاه آزاد اسلامی، کرمان، ایران
کلید واژه: تنوع ژنتیکی, genetic diversity, لاکتوباسیلوس, Lactobacilli, RAPD-PCR, درخت فیلوژنی, Phylogeny Tree,
چکیده مقاله :
محصولات لبنی مناطق مختلف می توانند منبعی برای سویه های جدید لاکتوباسیلوس باشند، بنابراین شناسایی مولکولی و بررسی تنوع ژنتیکی آنها می تواند گام موثری در شناسایی ذخیره لاکتوباسیلوس های بومی با خصوصیات عملکردی ویژه و به کارگیری آنها در محصولات لبنی صنعتی شود. هدف از این تحقیق جداسازی لاکتوباسیلوس ها از فلور موجود در شیر گاو شهرستان نرماشیر و بررسی تنوع ژنتیکی آنها می باشد. بدین منظور تعداد 21 نمونه شیر از مناطق مختلف این شهرستان جمع آوری و با روش های مرسوم فنوتیپی و بیوشیمیایی لاکتوباسیلوس ها جداسازی شدند. به منظور غربالگری و بررسی تنوع ژنتیکی لاکتوباسیلوس های جدا شده، پنج پرایمر RAPD به طور تصادفی انتخاب گردید و پس از PCR، تجزیه و تحلیل تنوع ژنتیکی و فیلوژنتیکی با استفاده از نرم افزار Pyelph انجام و درخت فیلوژنی رسم شد. تجزیه خوشه ای داده های مولکولی توانست جدایه ها را در فاصله ژنتیکی 10 به هفت گروه مجزا تقسیم بندی کند. جدایه های K3 و K4 در گروه یک و جدایه های K21 و K13 و K11 در گروه چهار قرار گرفتند. بقیه جدایه ها در گروه های مجزا قرار گرفتند. از آنجائیکه جدایه های K3 و K4 از یک ناحیه و جدایه های K21 و K13 نیز از یک ناحیه جمع آوری شده اند، قرار گرفتن آنها در یک گروه منطقی به نظر می رسد. نتایج بدست آمده از این تحقیق بیانگر تنوع ژنتیکی بالای جدایه های بومی شهرستان نرماشیر و مناسب بودن تکنیک RAPD-PCR برای مطالعات مشابه می باشد.
The use of lactobacilli in the production of fermented foods has several thousand years old due to their ability to make the desired changes in the taste and texture of the food, as well as their importance in the human health. Different dairy products can be a source of new strains of Lactobacillus. Therefore, molecular identification and their genetic variation can be an effective step in identifying native lactobacilli reserves with specific functional characteristics and applying them to industrial dairy products. The purpose of this study is to isolate lactobacilli from flora in cow's milk of Narmeshir city and study their genetic diversity. For this purpose, 21 milk samples were collected from different regions of the city, and Lactobacilli wereisolated by the conventional phenotypic and biochemical methods. In order to screen and investigate the genetic diversity of isolated lactobacilli, five RAPD primers were randomly selected. After doing PCR, the analyses of genetic and phylogenetic diversity were performed using Pyelph software and phylogeny trees were drawn. Cluster analysis of molecular data was able to divide the isolates into seven distinct groups at a genetic distance of 10. K3 and K4 isolates were in group 1 and isolates K21, K13, and K11were in group 4. The remaining isolates were placed in separate groups. Since K3 and K4 isolates were from a region and K21 and K13 isolates were also collected from a region, their placement seems to be logical in a group.
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