بررسی سرولوژیکی و مولکولی ویروس موزائیک معمولی لوبیا در استان زنجان

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی‌ارشد بیماری‌شناسی گیاهی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه زنجان، زنجان

2 استادیار گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه زنجان، زنجان

3 استادیار پژوهش، مؤسسه تحقیقات اصلاح و تهیه نهال و بذر، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج

4 استادیار گروه بیوتکنولوژی، پژوهشکده فناوری‌های نوین زیستی دانشگاه زنجان، زنجان

چکیده

ویروس موزائیک معمولی لوبیا (BCMV Bean common mosaic virus,) از مهم‌ترین عوامل بیماریزای لوبیا در سراسر جهان می‌باشد. این تحقیق به‌منظور ردیابی این ویروس از مزارع لوبیای استان زنجان انجام شد. برای این منظور، تعداد 130 برگ از نمونه‌های لوبیا با علایم موزائیک، رگبرگ نواری، بد شکلی و قاشقی شدن از مزارع استان زنجان در سال 1393 جمع‌آوری و توسط آزمون سرولوژیکی الایزا (DAS- ELISA) با استفاده از آنتی‌بادی چند همسانه‌ای، آلودگی آنها به ویروس تأیید شد. جهت خالص‌سازی بیولوژیک، ویروس روی گیاه محک و سپس روی لوبیای رقم اختر مایه­زنی و تکثیر شد. سپس با استفاده از آغازگرهای اختصاصی طراحی شده براساس توالی پروتئین پوششی، قسمتی از ژنوم ویروس تکثیر و توسط شرکت ماکروژن توالی‌یابی گردید. براساس نتایج آزمون الایزا بیش‌ترین میزان آلودگی نمونه­های جمع­آوری شده مربوط به نمونه‌های مزارع دانشگاه زنجان و کمترین میزان آلودگی در مزارع سوکهریزی و نعلبندان مشاهده شد. نتایج مقایسه توالی بخشی از پروتئین پوششی با توالی سایر ویروس‌ها نشان داد که جدایه غالب این ویروس به جدایه ای از آفریقای جنوبی با رس شمار AF361337 شباهت بالایی (99%) دارد. همچنین ترسیم درخت فیلوژنتیک مربوط به توالی تعیین شده نشان داد، این جدایه ویروسی با سایر جدایه‌های گزارش شده از ایران در یک خوشه قرار می‌گیرند که بیانگر تنوع پایین این ویروس در یک محدوده خاص جغرافیایی است.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Serological and Molecular Study of Bean Common Mosaic Virus in Zanjan Province

نویسندگان [English]

  • Elham Ahangar Moghtader 1
  • Omid Eini Gandomani 2
  • Rahim Ahmadvand 3
  • Abbas Bahari 4
1 M.Sc. Graduated of Plant Pathology, Department of Plant Protection, Faculty of Agriculture, University of Zanjan, Zanjan
2 Assistant Professor, Department of Plant Protection, Faculty of Agriculture, University of Zanjan, Zanjan
3 Research Assistant Professor, Seed and Plant Improvement Institute (SPII), Agricultural Research, Education and Extension Organization (AREEO), Karaj
4 Assistant Professor, Department of Biotechnology, Research Institute of Modern Biological Techniques, University of Zanjan, Zanjan
چکیده [English]

Bean common mosaic virus (BCMV) is one of the most important diseases that reduce the yield of common bean worldwide. To detect and identify of BCMV from common bean fields in Zanjan province, 130 samples were collected that showing mosaic, vein stripe, chlorotic spot and leaf deformation symptoms in 2014. BCMV infection in these samples was assayed by DAS-ELISA using polyclonal antibody. Indicator plant, Chenopodium quinoa, was used to purify virus biologically and then pathogenicity test was carried out on susceptible bean variety, Akhtar. A fragment of viral genome was amplified by polymerase chain reaction using specific primers designed for a part of the coat protein gene and then sequenced by Macrogen Company. The results of ELISA test showed the highest disease incidence in collected samples from Zanjan University fields and lowest number was observed in Sookakrizi and Nalbandan. Moreover, the comparison of the obtained sequence with other viruses in GenBank database showed the highest similarity (99%) with a BCMV isolate with Genbank accession number AF361337 from South Africa. A phylogenetic tree was also constructed by MEGA6 software. This isolate was grouped in the same clade as other isolates from Iran which indicates a low genetic variation for this virus in an especial geographical region.

کلیدواژه‌ها [English]

  • Potyvirus
  • Common bean
  • ELISA
  • Zanjan province
  • BCMV

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