نخستین بررسی ویروئید پوست پینه‏ای سیب در میزبان‌های سیب و گلابی در ایران و تعیین میزان تنوع ژنتیکی جدایه‏های ایرانی

نوع مقاله : مقالات پژوهشی

نویسندگان

1 دانشگاه فردوسی مشهد

2 دانشگاه آدلاید

چکیده

ویروئید پوست پینه‏ای سیب، ASSVd(Apple scar skin viroid)، یکی از مخرب‌ترین بیماری‌های ویروئیدی در میان درختان میوه دانه‏دار بوده و میتواند خسارات اقتصادی چشم‌گیری ایجاد نماید. علایم این بیماری شامل شکاف خوردگی، بدشکلی و خال‌دار شدن میوه در باغات سیب و گلابی در استان خراسان رضوی مشاهده گردید. نمونه‌برداری از باغات سیب و گلابی صورت گرفته و RNA از برگ‌های درختان با استفاده از روش به دام اندازی سلیکا (Silica capture method) استخراج گردید. ASSVd پس از انجام آزمون RT-PCR در نمونه‌های حاصل از این باغات شناسایی شد. محصول PCR خالص گردیده و با استفاده از ناقلpGEM®-T Easy همسانه‏سازی شده و پلاسمید نوترکیب حاصل توالی یابی شد. پس از انجام آنالیزهای لازم، 12 جدایه جدید این ویروئید از میزبان‌های سیب و گلابی در بانک ژن (NCBI) ثبت گردید. این جدایه‏ها‏ طولی بین 334 -329 نوکلئوتید داشتند. آنالیز فیلوژنتیکی با استفاده از روش neighbor joining نشان داد که جدایه‏های ایرانی از سایر جدایه‏های این ویروئید مشخص بوده و در گروه جداگانه‏ای قرار گرفتند. تنوع ژنتیکی از طریق نشان دادن جایگاه‌های جهش جدایه‏های ایرانی بر روی ساختار ثانویه مولکول مرجع ASSVd مورد بررسی قرار گرفت. این مطالعه اولین گزارش از شناسایی و بررسی صفات ویروئید پوست پینه‏ای سیب در ایران میباشد.

کلیدواژه‌ها

عنوان مقاله [English]

First Characterization of Apple scar skin viroid from Apple and pear cultivars in Iran and determination of genetic diversity

نویسندگان [English]

  • A. Yazarlou 1
  • B. Jafarpour
  • M. Koohi Habibi
  • S. Tarighi 1
  • J. W. Randles 2
1 Ferdowsi University of Mashhad
2 University of Adelaide
چکیده [English]

Apple scar skin viroid (ASSVd) is one of the most destructive viroid diseases in pome fruits orchards which can cause significant economic losses. Symptoms of this viroid including scaring, distortion and dappling were seen in some apple orchards in east-northern of Iran. Sampling was carried out from apple and pear orchards. The RNA was extracted by Silica capture method from leaves. ASSVd was detected by RT-PCR. The purified PCR products were cloned into pGEM®-T Easy Vector and recombinant plasmids were sequenced and sequencing analysis was done. We reported 12 new variants of ASSVd from apple and pear hosts with variable length between 329 to 334 nucleotides. Phylogenetic analysis by the neighbour joining method showed that the Iranian isolates were distinct from other isolates of ASSVd. Mutation sites were indicated along the secondary structure of ASSVd molecule. This study provides the first report of detection and molecular characterization of ASSVd in apple and pear in Iran.

کلیدواژه‌ها [English]

  • ASSVd
  • Secondary structure
  • Cloning
  • Phylogenetic analysis

مراجع

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