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شناسایی و بررسی برخی از ویژگی‌های ویروس آلوده‌کنندة زعفران (Crocus sativus) در ایران

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

2 دانشیار گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

3 استاد گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

4 دانشیار گروه باغبانی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

5 دانشجوی سابق گروه باغبانی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

چکیده

پس از نمونه‌برداری تصادفی از برگ‌ها و بنه‌های گیاه زعفران (Crocus sativus L.) از شش استان کشور در سال‌های 1390 تا 1394، شمار 641 از 890 نمونه با آنتی‌بادی­های عمومی جنس پوتی‌ویروس و همان‌طور با آنتی­بادی اختصاصی ویروس موزاییک معمولی لوبیا(BCMV)  در آزمون الایزا واکنش مثبت نشان داد. وزن مولکولی پروتئین پوششی (CP) چند جدایة مورد بررسی پوتی‌ویروس زعفران با استفاده از آزمون وسترن بلات 35 کیلو دالتون محاسبه شد. پس از استخراج آر.ان.ا کل از گیاهان آلوده، سه قطعه منقطع از ژنوم (مربوط به انتهای '3 ژنوم و بخشی از نواحی CI و HC-Pro) تکثیر شد. پروتئین پوششی (CP) پوتی‌ویروس زعفران بر پایة توالی نوکلئوتیدی بیشترین یکسانی را (74 درصد) با Ceratobium mosaic virus و Telosma mosaic virus  و بر پایة  توالی ترجمة اسیدآمینه‌ای بیشترین یکسانی را (8/78 درصد) با Bean common mosaic necrosis virus  داشت. به‌رغم رابطة سرم‌شناختی (سرولوژیکی) بالای این پوتی‌ویروس با BCMV و خویشاوندی تبارزایی (فیلوژنتیکی) با زیرگروه BCMV بر پایة ناحیة CP، میزان یکسانی توالی نوکلئوتیدی و ترجمه اسید آمینه­ای پروتئین پوششی آن کمتر از آستانة تعیین حدود و گسترة گونه در جنس پوتی­ویروس است. لذا به‌احتمال‌زیاد پوتی‌ویروس آلوده‌کنندة زعفران گونة جدیدی از جنس‌ پوتی‌ویروس بوده که اظهارنظر قطعی مستلزم تعیین توالی کل ژنوم آن است.

کلیدواژه‌ها


عنوان مقاله [English]

Identification and partial characterization of the virus infecting saffron (Crocus sativus) in Iran

نویسندگان [English]

  • Shirin Parizad 1
  • Akbar Dizadji 2
  • Mina Koohi Habibi 2
  • Gholam Hossein Mosahebi Mohammadi 3
  • Siamak Kalantari 4
  • Fatemeh Izadpanah 5
1 Ph.D. Student, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
2 Associate Professor, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3 Professor, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
4 Associate Professor, Department of Horticultural Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
5 Former M.Sc. Student, Department of Horticultural Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
چکیده [English]

Following the random sampling of saffron (Crocus sativus L.)leaves and corms from six provinces of Iran, during growing seasons of 2011-2015, a total of 890 leaf tissue samples were investigated by enzyme-linked immunosorbent assay (ELISA) tests. The results revealed that 641 samples showed positive reaction only with Potyvirus genus specific and exactly with Bean common mosaic virus (BCMV) specific antibodies. The molecular mass of several isolates of the virus coat protein (CP) was estimated as 35 kDa by western blotting. Following total RNA extraction from virus infected samples, three discontinuous fragments (corresponding to genomic 3', partial CI and HC-Pro regions) were amplified. The CP region of the virus infecting saffron has the highest nucleotide identity with Ceratobium mosaic virus (CerMV) and Telosma mosaic virus (TelMV) (74%) and the highest deduced amino acid identity with Bean common mosaic necrosis virus (BCMNV) (78.8%). Despite the high serological relationship with BCMV and a close phylogenetic relationship with BCMV subgroup based on CP region, the identity values are less than described species demarcation criteria for Potyviridae family. Overall, it seems that the saffron infecting Potyvirus is a new species of Potyvirus genus but the definitive statement requires sequencing the entire genome of the virus.

کلیدواژه‌ها [English]

  • Coat protein
  • ELISA
  • Potyvirus
  • saffron (Crocus sativus L.)
Adams, M. J., Antoniw J. F. & Fauquet C. M. (2005a). Molecular criteria for genus and species discrimination within the family Potyviridae. Archives of Virology, 150 (3), 459-479.

  • Adams, M. J., Antoniw, J. F. & Beaudoin, F. (2005b). Overview and analysis of the polyprotein cleavage sites in the family Potyviridae. Molecular Plant Pathology, 6,471-487.

  • Brunt, A. A. & Phillips, S. (1979). Viruses of bulb crops. Crocus tomasinianus and Narcissus spp. Annual Report of the Glasshouse Crops Horticultural Research Institute, 130-131.

  • Chen, C. C., Chang, C. A., Tsai, H. T. & Hsu, H. T. (2004). Identification of a Potyvirus causing latent infection in calla lilies. Plant Disease, 88, 1046.

  • Chen, J. & Chen, J. S. (2000). Occurrence and control of mosaic disease (Turnip mosaic virus) in saffron (Crocus sativus). Zhejiang Nongye Kexue, 3, 132-135.

  • Clark, M. F. & Adams, A. N. (1977). Characteristics of microplate method of enzyme linked immunosorbent assay for the detection of plant viruses. Journal of General Virology, 34, 475-483.

  • Fried, M. & Crothers, D.M. (1981). Equilibria and kinetics of lac repressor–operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Research, 9, 6505-6525.

  • Grilli Caiola, M. & Faoro, F. (2011). Latent virus infections in Crocus sativus and Crocus cartwrightianus. Phytopathologia Mediterranea, 50, 175-182.

  • Grilli Caiola, M. (1982). Virus-like particles in cells of saffron flowers. Phytopathology Zeitschrift, 105, 92-95.

  • Ha, C., Coombs, S., Revill, P. A., Harding, R. M., Vu, M. & Dale, J. L. (2008). Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses. Archives of Virology, 15, 25-36.

  • Huang, X. & Miller W. (1991). A time-efficient linear-space local similarity algorithm. Advances in Applied Mathematics, 12, 337-357.

  • Hull, R. (2002). Matthews' Plant Virology (4th ed.), New York, USA. Academic Press. 1001 pp.

  • Jensen, S. G., Long-Davidson, B. & Seip, L. (1986). Size variation among proteins induced by sugarcane mosaic viruses in plant tissue. Phytopathology, 76, 528-532.

  • JiShuang, C. (2000). Occurrence and control of mosaic disease (Turnip mosaic virus) in saffron (Crocus sativus). Journal Zhejiang Nongye Kexue, 3, 132-135.

  • Joisson, C., Dubs, M. C., Briand, J. P., Van Regenmortel, M., H. (1992). Detection of potyviruses with antisera to synthetic peptides. Research in Virology, 143 (3), 167-178.

  • Jones, D. T., Taylor, W. R., Thornton, J. M. (1992). The rapid generation of mutation data matrices from protein sequences. Computer Applications in the Biosciences, 8, 275-282.

  • King, A. M. Q., Adams, M. J., Carstens, E. B. and Lefkowitz, E. J. (2011). Taxonomy: Classification and Nomenclature of Viruses: Ninth Report of the International Committee on Taxonomy of Viruses. San Diego. Elsevier, 1327 pp.

  • Lapierre, H. & Singnoret, P. A. (2004). Viruses and virus disease of Poaceae (Gramineae). USA. Science publishers.

  • Leammli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Journal of Nature, 277, 680-685.

  • Mackenzie, A. M., Nolan, M., Wei, K. J., Clements, M. A., Gowanlock, D., Wallace, B. J. & Gibbs, A. J. (1998). Ceratobium mosaic potyvirus: another virus from orchids. Archives of Virology, 143, 903-914.

  • Miglino, R., Jodlowska, A. & van Schadewijk, A. R. (2005). First report of Narcissus mosaic virus Infecting Crocus spp. cultivars in the Netherlands. Plant Disease. 89 (3), 342.

  • Ochwo-Ssemakula, M., Sengooba, T., Hakiza, J. J., Adipala, E., Edema, R., Redinbaugh, M. G., Aritua, V. & Winter. S. (2012). Characterization and distribution of a potyvirus associated with passion fruit woodiness disease in Uganda. Plant Disease, 96, 659-665.

  • Pisi, A. & Bellardi, G. (1990). Ultrastructural study of cytoplasmic inclusions in plants infected with Potyviruses. Journal of Phytopathology, 130, 114-118.

  • Russo, M., Martelli, G. P., Cresti, M. & Ciampolini, F. (1979). Bean yellow mosaic virus in saffron. Phytopathologia Mediterranea, 18, 189-191.

  • Sambrook, J. & Russell, D. (2001). Molecular Cloning: A Laboratory Manual (3rd ed.). New York, USA: Cold Spring Harbor Laboratory Press. 2100 pp.

  • Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (3rd ed.). New York, USA: Cold Spring Harbor Laboratory Press. 1626 pp.

  • Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013). MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30, 2725-2729.

  • Tavaré, S. (1986). Some Probabilistic and Statistical Problems in the Analysis of DNA Sequences (PDF). Lectures on Mathematics in the Life Sciences. American Mathematical Society, 17: 57-86.

  • Towbin, H., Staehelin, T. & Gordon, J. (1997). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Academic Science USA, 76, 4350-4354.

  • Urcuqui-Inchima, S., Haenni, A. L. & Bernardi, F. (2001). Potyvirus proteins: a wealth of functions. Virus Research, 74, 157-175.

  • Van Slogtern, D. H. M. (1958). Rattle virus as a cause of diseases in flower bulbs, and the possibility of controlling infection by soil disinfectants. Tijdschr PlZiekt, 64, 5-6.