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شناسایی مولکولی چهار ویروس مهم جنس نپوویروس از تاکستان‌های استان زنجان

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق کارشناسی ارشد، گروه گیاهپزشکی، دانشکدۀ کشاورزی، دانشگان زنجان

2 استادیار، گروه گیاهپزشکی، دانشکدۀ کشاورزی، دانشگان زنجان

3 استادیار، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان. سنندج

چکیده

به­منظور بررسی وضعیت ویروس­های مهم متعلق به جنس نپوویروس از تاکستان­های استان زنجان، نمونه‌برداری در سال­های زراعی 95-۹4 به‌صورت انتخابی از گیاهان دارای نشانه‌های مشکوک به بیماری‌های ویروسی انجام و درمجموع 168 نمونۀ گیاهی دارای نشانه و بدون نشانه گرد‌آوری شد. استخراج آر.ان.­ای کل از بافت­های برگی و آوندی 57 نمونه بر اساس نشانه‌ها و مناطق نمونه‌برداری شده انجام گرفت و cDNA آن­ها ساخته‌ شد و سپس، آزمون پی­سی­آر توسط آغازگرهای اختصاصی برای تکثیر (افزونش) بخشی از ژنگان (ژنوم) نپوویروس‌های موزاییک آرابیس (Arabis mosaic virus, ArMV)، بدشکلی انگور (Grapevine deformation virus, GDeV)، برگ بادبزنی انگور (Grapevine fanleaf virus, GFLV) و لکۀ حلقوی گوجه­فرنگی (Tomato ring spot virus, ToRSV) انجام گرفت. نتایج واکنش زنجیره­ای پلمیراز گویای تکثیر قطعه‌های مورد انتظار از ویروس‌­های ArMV، GDeV، GFLV و ToRSV به ترتیب از 7/15، 7/15، 5/10 و 14 درصد نمونه­ها بود. پس از تعیین توالی، همردیف‌سازی ترادف نوکئوتیدی قطعه‌های تکثیرشده با دیگر توالی‌های موجود در بانک ژن و تحلیل تبارزائی توسط برنامۀ MEGA6، با روش Neighbor-Joining انجام شد. بررسی رابطه‌های تبارزائی بر اساس توالی­های نوکلئوتیدی نشان داد، جدایه­های ردیابی­شده در این تحقیق با جدایه­هایی از کشورهای مختلف هم­گروه‌اند که در بیشتر موارد ارتباط جغرافیایی بین جدایه‌های ردیابی شده و دیگر جدایه‌ها را نشان می‌دهد. این نخستین تحقیق جامع بر اساس داده­های مولکولی از نپوویروس­های مهم انگور در استان زنجان و نخستین گزارش ویروس بدشکلی انگور از این منطقه است.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular Identification of four important nepovirus from vineyards of Zanjan province

نویسندگان [English]

  • Roghayeh Moradi 1
  • Davoud Koolivand 2
  • Omid Eini 2
  • Mohammad Hajizadeh 3
1 Former M. Sc. Student, Department of Plant Pathology, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
2 Assistant Professor, Department of Plant Pathology, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
3 Assistant Professor, Department of Plant Protection, Faculty of Agriculture, University of Kurdistan, Sannadej, Iran
چکیده [English]

To determine incidence of nepoviruses in vineyards of zanjan province, 168 symptomatic and non-symptomatic samples were collected during season growing 2014-2015. Total RNA was extracted from the 57 leaves and green shoots of selected samples and then cDNAs were synthesized. Genomic segment of Arabis mosaic virus (ArMV), Grapevine deformation virus (GDeV), Grapevine fan leaf virus (GFLV) and Tomato ring spot virus (ToRSV) was amplified by polymerase chain reaction (PCR) using specific primers. The result revealed that the expected bands were amplified at 15.7, 15.7, 10.5 and 14% of different samples belong to ArMV, GDeV, GFLV and ToRSV, respectively. Following sequencing and multiple alignment of nucleotide sequence of amplified fragments with isolates that retrieved from NCBI. Phylogenetic tree was created by MEGA6 software using Neighbour-Joining method. The result revealed that the Iranian isolates were grouped with reported isolates from different regions around the world based on geographical. This is the first comprehensive study based on molecular methods to determine grapevine nepoviruses in Zanjan province and the first report of GDeV from Zanjna based on our knowledge.

کلیدواژه‌ها [English]

  • Arabis mosaic virus
  • grapevine deformation virus
  • grapevine fanleaf virus
  • RT-PCR
  • tomato ringspot virus
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