شناسایی مولکولی ویروس پیسک توت‌فرنگی (Strawberry mottle virus) از مزارع توت‌فرنگی استان کردستان بر اساس منطقه 3΄NCR

نوع مقاله: بیماری‌شناسی گیاهی

نویسندگان

1 گروه گیاهپزشکی دانشگاه کردستان، سنندج، ایران

2 هیات علمی دانشگاه کردستان

10.22092/jaep.2019.125077.1266

چکیده

در تابستان 1397، علائم ویروسی شامل پیسک، بدشکلی و سوختگی حاشیه برگ­ها از مزارع توت­فرنگی استان کردستان مشاهده و تعداد 48 نمونه برگی جمع­آوری شد. ابتدا برخی نمونه­ها به گیاه محک Fragaria vesca با استفاده از شته توت­فرنگی (Chaetosiphon fragaefolii) تلقیح و علایم مطابق با Strawberry mottle virus (SMoV) ظاهر شد. سپس، آر ان ­ا کل از نمونه­ها با روش سیلیکا استخراج و دی ان ا مکمل آن‌ها با آغازگرهای تصادفی شش­تایی ساخته شد. PCR با آغازگرهای اختصاصی SMoV- Sm2ncr1a و SMoV-Smdetncr4a انجام و قطعه­ای به طول 461 جفت باز در 28 نمونه از 48 نمونه فوق تکثیر گردید. به‌منظور اطمینان از نتیجه RT-PCR و بررسی مشخصات مولکولی، پنج جدایه براساس منطقه جغرافیایی، رقم میزبان و نوع علایم انتخاب و پس از الحاق به پلاسمید pTG-19 در باکتری Escherichia coliانتقال و پلاسمیدهای نوترکیب همسانه شده تعیین توالی شدند. میانگین فاصله ژنتیکی در بین جدایه­های در این تحقیق در سطح نوکلئوتیدی 002/0 ± 005/0 و با سایر جدایه­های موجود در بانک ژن 008/0 ± 084/0 بود. مقایسه دوبه دوی توالی­های نوکلئوتیدی نشان داد که بیشترین و کمترین فاصله ژنتیکی به‌ترتیب میان جدایه K3 با جدایه NSper51 از کانادا (015/0 ± 093/0) و جدایه SQA یا AG با جدایه NB926 از کانادا (007/0 ± 019/0) وجود داشت.این اولین گزارش از شناسایی این ویروس در مزارع توت­فرنگی استان کردستان می­باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular identification of Strawberry mottle virus in strawberry fields of Kurdistan Province based on the 3΄NCR

نویسندگان [English]

  • Nasrin Ghaderi Zandan 1
  • Mohammad Hajizadeh 2
1 Plant Protection Department, University of Kurdistan, Sanandaj, Iran
چکیده [English]

In the summer of 2018, viral symptoms corresponded to Strawberry mottle virus (SMoV) in some strawberry fields were observed and 48 leaf samples/plants were collected from different fields from Kurdistan Province. In the preliminary test, some samples were inoculated to Fragaria vesca by strawberry aphid (Chaetosiphon fragaefolii) and SMoV-symptoms were observed after about two weeks. Then, total RNA of samples was extracted by silica-capture method and subjected to cDNA synthesize with random hexamer primers. RT-PCR was done by specific SMoV pair primer and amplified fragments were observed on 1.2% agarose gels. RT-PCR results were showed that 28 out of 48 samples were infected by SMoV. Based on the geographical origin, cultivar and symptoms, five isolates were selected, ligated to pTG-19 and transformed to Escherichia coli. Recombinant plasmids were sequenced and analyzed for molecular characterization and phylogenetic studies on SMoV 3΄NCR. Mean genetic distance between the isolates from Iran was 0.005 ± 0.002 but between these isolates and the other isolates available in GenBank was 0.084 ± 0.008. The highest and lowest genetic distance was found between K3 with NSper51 (0.093 ± 0.008) and SQA/AG with NB926 (0.019 ± 0.017) from Canada, respectively. This is the first report on occurrence of SMoV on strawberry in Kurdistan Province.
 

کلیدواژه‌ها [English]

  • Genetic distance
  • phylogeny
  • Strawberry
  • viral infectio

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