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ردیابی همزمان دو گونه ویروس رگبرگ زرد نکروتیک چغندرقند و ویروس سوختگی سیاه چغندرقند با استفاده از واکنش زنجیرهای پلیمراز دوگانه | ||
گیاه پزشکی | ||
دوره 46، شماره 1، اردیبهشت 1402، صفحه 9-23 اصل مقاله (732 K) | ||
نوع مقاله: علمی پژوهشی-فارسی | ||
شناسه دیجیتال (DOI): 10.22055/ppr.2023.18033 | ||
نویسندگان | ||
جمشید سلطانی ایدلیکی* 1؛ محسن مهرور2 | ||
1استادیار، بخش تحقیقات چغندرقند، مرکز تحقیقات و آموزش و منابع طبیعی خراسان رضوی، سازمان تحقیقات ،آموزش و ترویج کشاورزی، مشهد، ایران | ||
2دانشیار، بخش گیاهپزشکی،دانشکده کشاورزی دانشگاه فردوسی ، مشهد، ایران | ||
چکیده | ||
تاکنون ویروسهای رگبرگ زرد نکروتیک چغندرقند (Beet necrotic yellow vein virus, BNYVV) و سوختگی سیاه چغندرقند (Beet black scorch virus, BBSV) بهطور مجزا در ریشههای آلوده از مزارع چغندرقند ایران ردیابی شدهاند. هدف از این مطالعه ردیابی همزمان دو ویروس ذکرشده بهمنظور صرفهجویی در زمان و انرژی در فرآیند تشخیص آلودگی بود. بدین منظور از سه روش استفاده شد. در روش اول (روش رایج): ویروسها با استفاده از واکنش زنجیرهای پلیمراز با ترانویسی معکوس (RT-PCR) بهوسیله آغازگرهای اختصاصی مربوط به هر کدام و مجزا از یکدیگر روی ریشه آلوده ردیابی شدند. در روش های دوم و سوم، این دو ویروس به ترتیب با استفاده از واکنش زنجیرهای پلیمراز با ترانویسی معکوس بهصورت تکی (Simplexrt-PCR) اما همزمان در یک ترموسایکلر و به صورت دوگانه (DuplexRT-PCR) با به کارگیری دو جفت آغازگر اختصاصی ردیابی شدند. نتایج در روش های دوم و سوم نشان داد که با بهینهسازی دمای اتصال آغازگرهای اختصاصی (56 درجه سانتیگراد) مبتنی بر ناحیه ژن کد کننده پروتئین پوششی، به ترتیب قطعات هدف 391 و 453 جفت بازی از ژنوم BNYVV و BBSV تکثیر مییابند. در نتیجه این دو روش جهت ردیابی همزمان مناسب بوده و ازنظر صرفهجویی در زمان و انرژی مقرونبهصرفه است و همچنین قابلیت استفاده از آنها بهمنظور تهیه نقشه پراکنش در مناطق کشت چغندرقند و نیز ارزیابی و شناسایی لاین های مقاوم چغندرقند در برنامههای به نژادی در شرایط گلخانه وجود دارد. | ||
کلیدواژهها | ||
آغازگر؛ ریزومانیا؛ ویروس سوختگی سیاهِ چغندرقند؛ واکنش زنجیرهای پلی مراز همراه با ترانویسی معکوس | ||
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