شناسایی جدایه های Pseudomonas syringae pv syringae جدا شده از درختان میوه هسته دار با استفاده از انگشت نگاری ژنتیکی با توالی REP
محورهای موضوعی : گیاه پزشکیساغر کتابچی 1 , نادر حسن زاده 2
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کلید واژه: rep-PCR, rep-PRC, Pseudomonas syringae, درختان میوه هسته دار, stone fruits trees,
چکیده مقاله :
در این تحقیق نقوش ژنومی rep-PCR مربوط به 24 جدایه P. syringae pv. syringae که از نواحی مختلف ایران و از روی میزبان های مختلف جداسازی شده بودند در مقایسه با استرین استاندارد از کشور یونان مورد ارزیابی قرار گرفتند. DNA باکتری های مورد مطالعه به سه روش فریز- جوشاندن، استفاده از کلنی های خالص و از طریق شستشوی برگ حاوی باکتری تهیه شد. نتایج بدست آمده نشان داد در روش فوق نیاز به استخراج DNA نیست. به علاوه بررسی انگشت نگاری REP نشان داد خصوصیات میزبان و محل زندگی آنها بر روی ژنوم باکتری تاثیر متقابل دارند. به طوری که جدایه هایی که از درختان میوه هسته دار جدا شده بودند مجموعه ای بزرگ و مستقل از جدایه های گندم و نیشکر را تشکیل می دادند. همچنین جدایه های درختان میوه هسته دار که مربوط به یک منطقه جغرافیائی بودند از شباهت های بیشتری بر خوردار بودند. استفاده از توالی REP در روش rep-PCR به عنوان یک روش ساده، دقیق و سریع در جهت شناسائی و طبقه بندی ایزولههای پاتوارهای P. syringae مورد مطالعه و تائید قرار گرفت.
Repetitive PCR fingerprinting of 24 strains of P. syringae pv. syringae isolated from different host/areas were compared with a standard strain of Pss from Greece (BPIC 242). Bacterial DNA was prepared with three known methods i.e. freeze-boil, whole colony boiling and directly from washed leaf surface. Initial result showed no purified genomic DNA extraction was necessary for rep-PCR analysis. On the other hand, Rep patherns of Pss showed some correlation between host specialization and its habitant with genomic diversity. Strains isolated from stone fruits formed a distinct cluster from wheat and sugar beet strains and a similar pathern was found among all stone fruits strains in confined geographical areas. Therefore it was found the rep-PCR technique as a rapid, simple and producible method to identify and classify these important plant pathogens.
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