تعیین مشخصات مولکولی و تبارزایی ویروئید لکه زردی-1 مو (Grapevine yellow speckle viroid-1) از تاکستان¬های شمال¬غرب کشور

نوع مقاله : مقاله پژوهشی


1 استادیار گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان

2 دانشیار گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه تبریز


به­منظور تعیین مشخصات مولکولی و بررسی ساختارهای ثانویه ویروئید لکه زردی- 1 مو، 137 نمونه از تاکستان­های استان­های اردبیل، آذربایجان­­­ شرقی و آذربایجان­ غربی جمع­آوری گردید. اسید نوکلئیک کل از بافت برگ نمونه­ها استخراج و پس از ساخت دی­ان­ای مکمل با استفاده از آغازگر شش­نوکلئوتیدی تصادفی، طول کامل این ویروئید با روش پی­سی­آر و با استفاده از آغازگرهای اختصاصی ویروئید لکه زردی -1 مو،  تکثیر شد. محصول پی­سی­آر پس از همسانه­سازی در حامل pGEMT-easy، تعیین­توالی گردید. نتایج نشان داد که واریانت­های شناسایی­شده در این تحقیق، دارای 367 یا 368 نوکلئوتید هستند و در مجموع هفت واریانت جدید از این ویروئید تشخیص داده شد. درخت تبارزایی نشان داد که واریانت 6 مشابه اعضای تیپ یک این ویروئید و شش واریانت بعدی در کنار تعدادی جدایه و واریانت دیگر تشکیل یک تیپ جداگانه (به­نام پیشنهادی تیپ 4) را می­دهند. در ساختار ثانویه واریانت­های به­دست­آمده در این تحقیق، بیشترین تفاوت در منطقه متغیر و قسمت انتهایی چپ قرار داشت. ویروئید لکه زردی-1 مو برای اولین بار از استان­های آذربایجان­شرقی و اردبیل گزارش می­شود.


عنوان مقاله [English]

Identification, Molecular and Phylogenetic Analyses of Grapevine yellow speckle viroid-1 from the Northwest Region of Iran

نویسندگان [English]

  • Mohammad Hajizadeh 1
  • Nemat Sokhandan Bashir 2
1 Assistant Professor, Dept of Plant Protection, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran
2 Associate Professor, Dept of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
چکیده [English]

Totally, 137 grapevines samples were collected from vineyards in East and West Azerbaijan and Ardabil provinces of Iran and tested for molecular characterization and study on the secondary structure of Grapevine yellow speckle viroid-1 (GYSVd-1). Total nucleic acid was extracted from leaves and first-strand cDNA synthesized using random hexamer primer. PCR fragments corresponding to full length GYSVd-1 were amplified by the use of specific primers and cloned in pGEM-T-easy for sequencing. Results showed that the Iranian isolates were 367-368 nt in length, and accordingly seven new GYSVd-1 variants were identified. Phylogenetic analysis revealed that variant 6 was identical to the type 1 of GYSVd-1, whereas the other six variants together with some other isolates and variants of GYSVd-1 formed a distinct clad, which named as GYSVd-1 type 4. These variants were most different from other isolates in variable and left terminal domains of their secondary structure. This is the first report of grapevine infection with GYSVd-1 from Ardabil and East Azarbaijan provinces.

کلیدواژه‌ها [English]

  • Grapevine
  • RT-PCR
  • Secondary structure
  • Variant
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