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mso-para-margin-top:0in mso-para-margin-right:0in mso-para-margin-bottom:10.0pt mso-para-margin-left:0in line-height:115% mso-pagination:widow-orphan font-size:11.0pt font-family:"Calibri","sans-serif" mso-ascii-font-family:Calibri mso-ascii-theme-font:minor-latin mso-fareast-font-family:"Times New Roman" mso-fareast-theme-font:minor-fareast mso-hansi-font-family:Calibri mso-hansi-theme-font:minor-latin mso-bidi-font-family:Arial mso-bidi-theme-font:minor-bidi} </style> <!--stripped--></div><p style="font-family: times new roman,times,serif text-align: justify" dir="rtl" class="BasicParagraph"><font size="2"><b><span lang="AR-YE" style="letter-spacing: 0.05pt">سابقه و هدف</span></b><span lang="AR-YE" style="letter-spacing: 0.05pt">: لاکتوباسیل ها متشکل از ارگانیزم های گرم مثبت، کاتالاز منفی، بدون اسپور و میله ای شکل هستند و بیش از 90 گونه از این باکتری شناخته شده است.<span> </span>سوش های متعددی از لاکتوباسیل های پروبیوتیک در طیف وسیعی از محصولات غذایی انسان ها وجود دارند. ظرفیت های پروبیوتیکی وابسته به سوش می باشند، انتخاب متدهای قابل اطمینان برای شناسایی لاکتوباسیل ها از اهمیت خاصی برخوردار است با توجه<span> </span>به<span> </span>وقت گیر بودن و ابهام آمیز بودن روش های بیوشیمیایی، روش های مولکولی مناسب تر و دقیق تر بوده و جایگزین روش های قبلی شده اند. هدف از این تحقیق، شناسایی لاکتوباسیل های جداشده از محصولات لبنی سنتی ایران با استفاده از مارکرهای </span><span style="letter-spacing: 0.05pt" dir="ltr">RAPD</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> به عنوان یک ابزار موثر در طبقه بندی لاکتوباسیل ها و بررسی تنوع ژنتیکی در<span> </span>سویه های جدا شده می باشد.</span></font></p><div style="text-align: justify"> </div><p style="font-family: times new roman,times,serif text-align: justify" dir="rtl" class="BasicParagraph"><font size="2"><b><span lang="AR-YE" style="letter-spacing: 0.05pt">مواد و روش ها</span></b><span lang="AR-YE" style="letter-spacing: 0.05pt">: در این مطالعه 20 ایزوله از محصولات لبنی مختلف جدا شدند و پس از کشت بر روی محیط اختصاصی </span><span style="letter-spacing: 0.05pt" dir="ltr">MRS</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> استخراج </span><span style="letter-spacing: 0.05pt" dir="ltr">DNA</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> از باکتری ها انجام شد و برای بررسی تنوع ژنتیکی از مارکرهای </span><span style="letter-spacing: 0.05pt" dir="ltr">RAPD</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> استفاده شد. </span></font></p><div style="text-align: justify"> </div><p style="font-family: times new roman,times,serif text-align: justify" dir="rtl" class="BasicParagraph"><font size="2"><b><span lang="AR-YE">یافته ها</span></b><span lang="AR-YE">: از 20 پرایمر به کار رفته در این تحقیق، 19 پرایمر باندهای قابل کد گذاری ایجاد کردند. تعداد کل باندهای تکثیر شده توسط کل پرایمرها، 225 عدد برآورد شد که شامل 219 باند پلی مورف، 6 باند مشترک و 63 باند اختصاصی بود. توسط نرم افزار<span> </span></span><span dir="ltr">NTSYS</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span>-</span><span dir="ltr">PC</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span> ماتریس تشابه بر اساس ضریب </span><span dir="ltr">Jaccard</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span> برای ژنوتیپ ها محاسبه شد. سپس گروه بندی ایزوله ها با روش </span><span dir="ltr">UPGMA</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span> و </span><span dir="ltr">NJ</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span> توسط نرم افزار </span><span dir="ltr">NTSYS</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span>-</span><span dir="ltr">PC</span><span dir="rtl"></span><span lang="AR-YE"><span dir="rtl"></span> انجام گرفت و نتایج یکسان و مشابهی از مقایسه گروه بندی با دو روش مختلف حاصل شد.<span style="letter-spacing: 0.05pt"></span></span></font></p><div style="text-align: justify"> </div><p style="font-family: times new roman,times,serif text-align: justify" dir="rtl" class="BasicParagraph"><font size="2"><b><span lang="AR-YE" style="letter-spacing: 0.05pt">نتیجه گیری</span></b><span lang="AR-YE" style="letter-spacing: 0.05pt">: بر اساس ماتریس تشابه </span><span lang="DA" style="letter-spacing: 0.05pt" dir="ltr">k2l3 </span><span lang="AR-YE" style="letter-spacing: 0.05pt">و</span><span dir="ltr"></span><span lang="DA" style="letter-spacing: 0.05pt" dir="ltr"><span dir="ltr"></span> y1m4</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> بیشترین تشابه ژنتیکی را با یکدیگر نشان دادند و در هر دو دندروگرام با هم در یک گروه قرار گرفتند. در گروه بندی براساس روش </span><span style="letter-spacing: 0.05pt" dir="ltr">UPGMA</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> دو شاخه اصلی </span><span style="letter-spacing: 0.05pt" dir="ltr">A</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> و </span><span style="letter-spacing: 0.05pt" dir="ltr">B</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> وجود داشت. شاخه اصلی </span><span style="letter-spacing: 0.05pt" dir="ltr">A</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> به دو زیر گروه با ژنوتیپ های </span><span lang="CS" style="letter-spacing: 0.05pt" dir="ltr">c₆m₃</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span>،</span><span lang="CS" style="letter-spacing: 0.05pt" dir="ltr">y₁l₄</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span>،</span><span dir="ltr"></span><span lang="CS" style="letter-spacing: 0.05pt" dir="ltr"><span dir="ltr"></span> y₂f₃</span><span lang="AR-YE" style="letter-spacing: 0.05pt">و</span><span dir="ltr"></span><span lang="AR-YE" style="letter-spacing: 0.05pt" dir="ltr"><span dir="ltr"></span> </span><span lang="NL" style="letter-spacing: 0.05pt" dir="ltr">c₁d₂</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> و شاخه اصلی </span><span style="letter-spacing: 0.05pt" dir="ltr">B</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span> نیز به دو گروه با ژنوتیپ های </span><span lang="DA" style="letter-spacing: 0.05pt" dir="ltr">c₂h₁</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span>،</span><span dir="ltr"></span><span lang="DA" style="letter-spacing: 0.05pt" dir="ltr"><span dir="ltr"></span> d₃b₁</span><span dir="rtl"></span><span lang="AR-YE" style="letter-spacing: 0.05pt"><span dir="rtl"></span>،</span><span dir="ltr"></span><span lang="DA" style="letter-spacing: 0.05pt" 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mso-fareast-theme-font:minor-fareast mso-hansi-font-family:Calibri mso-hansi-theme-font:minor-latin mso-bidi-font-family:Arial mso-bidi-theme-font:minor-bidi} </style> <!--stripped--><font size="2" style="font-family: times new roman,times,serif"><b><span style="font-size: 14pt line-height: 120% color: black">Aim and background</span></b><b><span style="font-size: 12pt line-height: 120% color: black">. </span></b><span style="font-size: 12pt line-height: 120% color: black">The genus Lactobacillus encompasses a diverse assemblage of Gram-positive, catalase-negative, non-spore-forming, rod-shaped organisms. Over 90 species are currently described. Several strains of probiotic lactobacilli have been incorporated in a wide range of products for human. As the probiotic capacities are strain-dependent, methods for reliable identification of lactobacilli at the strain level are of great importance. Regarding biochemical methods are time-consuming and often very ambiguous the molecular methods are more accurate. The aim of present study is typing of lactobacillus species isolated from different traditional dairy products by RAPD markers.</span> </font></div><p style="margin-bottom: 0.0001pt line-height: 120% vertical-align: middle font-family: times new roman,times,serif text-align: justify" class="MsoNormal"><font size="2"><b><span style="font-size: 14pt line-height: 120% color: black">Materials and Methods.</span></b><span style="font-size: 12pt line-height: 120% color: black"> The<b> </b>20<b> </b>strains<span> </span>used in this study were isolated from different sources. Samples were grown in MRS broth then total genomic DNA was extracted. RAPD markers were used for the detection of genetic diversity. </span></font></p><div style="text-align: justify font-family: times new roman,times,serif"> </div><p style="margin-bottom: 0.0001pt line-height: 120% vertical-align: middle font-family: times new roman,times,serif text-align: justify" class="MsoNormal"><font size="2"><b><span style="font-size: 14pt line-height: 120% color: black">Results.</span></b><span style="font-size: 12pt line-height: 120% color: black"> Out of 20 primers used 19 primers produced reproducible bands. The primers generated 225 RAPD loci, of which 219 were polymorph, 6 were monomorph and 63 were unique. NJ and UPGMA clustering methods performed on molecular data by NTSYS-PC software. Similar results are obtained by two methods. </span></font></p><div style="text-align: justify font-family: times new roman,times,serif"> </div><p style="margin-bottom: 0.0001pt line-height: 120% vertical-align: middle font-family: times new roman,times,serif text-align: justify" class="MsoNormal"><font size="2"><b><span style="font-size: 14pt line-height: 120% color: black">Conclusion</span></b><b><span style="font-size: 12pt line-height: 120% color: black">. </span></b><span style="font-size: 12pt line-height: 120% color: black">UPGMA clustering methods were performed on molecular data. Two genotypes of k₂l₃</span><span dir="rtl"></span><span style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span> </span><span style="font-size: 12pt line-height: 120% color: black">and</span><span dir="rtl"></span><span style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span> </span><span style="font-size: 12pt line-height: 120% color: black">y₁m₄</span><span dir="rtl"></span><span style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span> </span><span style="font-size: 12pt line-height: 120% color: black">are grouped together in two dendrograms. In UPGMA dendrogram two major clusters A and B were observed. Cluster A was divided in to2 subcluster by c₆m₃</span><span dir="rtl"></span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span>،</span><span style="font-size: 12pt line-height: 120% color: black">y₁l₄</span><span dir="rtl"></span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span>،</span><span dir="ltr"></span><span style="font-size: 12pt line-height: 120% color: black"><span dir="ltr"></span> y₂f₃ and</span><span dir="rtl"></span><span style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span> </span><span style="font-size: 12pt line-height: 120% color: black">c₁d₂ genotypes. In cluster B, 2 subclusters by 6 genotypes (c₂h₁</span><span dir="rtl"></span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span>، </span><span style="font-size: 12pt line-height: 120% color: black">d₃b₁</span><span dir="rtl"></span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span>، </span><span style="font-size: 12pt line-height: 120% color: black">y₂c₄</span><span dir="rtl"></span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl"><span dir="rtl"></span>،</span><span dir="ltr"></span><span style="font-size: 12pt line-height: 120% color: black"><span dir="ltr"></span> k₂l₃</span><span lang="FA" style="font-size: 12pt line-height: 120% color: black" dir="rtl">و </span><span style="font-size: 12pt line-height: 120% color: black">y₁m₄and p) were observed.</span></font></p><div style="text-align: justify font-family: times new roman,times,serif"> </div><p style="margin-bottom: 0.0001pt line-height: 120% vertical-align: middle font-family: times new roman,times,serif text-align: justify" class="MsoNormal"><font size="2"><b><span style="font-size: 14pt line-height: 120% color: black">Key words</span></b><b><span style="font-size: 12pt line-height: 120% color: black">.</span></b><span style="font-size: 12pt line-height: 120% color: black"> Lactobacillus strain, genetic diversity,</span><b><span style="font-size: 14pt line-height: 120% color: black"> </span></b><span style="font-size: 12pt line-height: 120% color: black">traditional dairy products, RAPD markers, UPGMA dendrogram.</span></font></p> لاکتوباسیل، محصولات لبنی سنتی، تنوع ژنتیکی، RAPD PCR، دندروگرام UPGMA Lactobacillus strain, genetic diversity, traditional dairy products, RAPD markers, UPGMA dendrogram 33 42 http://ncmbjpiau.ir/browse.php?a_code=A-10-2-18&slc_lang=fa&sid=1 Farzaneh Tafvizi فرزانه تفویضی farzaneh.tafvizi@yahoo.com 1003194753284600591 1003194753284600591 Yes Islamic Azad University, Parand Branch دانشگاه آزاد اسلامی واحد پرند Maryam Tajabadi ebrahimi مریم تاج آبادی ابراهیمی 1003194753284600592 1003194753284600592 No Islamic Azad University, Central Tehran Branch دانشگاه آزاد اسلامی واحد تهران مرکزی Mitra Heydari Nasrabadi میترا حیدری نصر آبادی 1003194753284600593 1003194753284600593 No Islamic Azad University, Parand Branch دانشگاه آزاد اسلامی واحد پرند Hoda Bahrami هدی بهرامی 1003194753284600594 1003194753284600594 No Islamic Azad University, Central Tehran Branch دانشگاه آزاد اسلامی واحد تهران مرکزی