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
Anonymous. 2014. Agricultural statistics Ministry of Jihad- E- Agriculture.
Bahrami Kamangar, S. 1998. Identification of Bean mosaic virus producing agents and relative frequency in Fars province. M.Sc. Thesis, University of Shiraz. pp 224. (In Persian).
Baradaran, G. H. 1997. Evaluation of Bean common mosaic virus (BCMV) and its distribution in Mashhad and Chenaran. M.Sc. thesis, University of Mashhad. pp 97.
Baradaran, G. H. and Jafarpour, B. 1998. Detection of Bean common mosaic virus and Cucumber mosaic virus in bean fields in Mashhad and Chenaran. 13th Iranian Plant Protection Congress. 157-163. (In Persian).
Clark, M. F. and Adams, A. N. 1977. Characteristics of microplate method of enzyme linked immunosorbent assay for detection of plant viruses. Journal of General Virology, 34: 475-483.
Dorri, H. R., Ghanbari, A. A., Lak, M. R. and Bani Jamali, M. 2008. Beans Guide. Publication of Agricultural Education. Karaj, 1: 124.
Drijfhout, E. 1978. Genetic interaction between Phaseolus vulgaris L. and Bean common mosaic virus with implications for strain identification and breeding for resistance. Agricultural Research Report 872, Pudoc, Wageningen.
Drijfhout, E. 1991. Bean Common Mosaic. Compendium of Bean Diseases. APS Press. pp 37-39.
Feng, X., Poplawsky, A. R., Nikolaeva, O. V., Myers, J. R., Karasev, A. V. 2014. Recombinants of Bean common mosaic virus (BCMV) and genetic determinants of BCMV involved in overcoming resistance in common bean. Phytopathology 104: 786-793.
Hashemi, M. 1999. The comparison of serological, biological and physico-chemical of Potyviruses Bean common mosaic virus, Bean yellow mosaic virus and Cowpea seed-borne virus in Fars. M.Sc. Thesis, University of Shiraz. pp 135. (In Persian).
Janssen, D., Martin, G., Velasco, L., Gomez, P., Segundo, E., Ruiz, L., and Cuadrado, I. M. (2005). Absence of a coding region for the helper component-proteinase in the genome of Cucumber vein yellowing virus, a whitefly-transmitted member of the Potyviridae. Archives of Virology, 150: 1439-1447.
Jooste, A. E. C., Spence, N. J., Van Tonder, T. and Pietersen, G. 2003. Pathogenicity of Bean common mosaic virus (BCMV) isolates collected from bean (P. vulgaris) in South Africa. Plant Virology, 08.
Kelly, J. D. 1997. A review of varietal response to Bean common mosaic potyvirus in Phaseolus vulgaris. Plant Varieties & Seeds, 10: 1-6.
Mavric, I. and Sustar-Vozlic, J. 2004. Virus diseases and resistance to Bean common mosaic and Bean common mosaic necrosis potyvirus in common bean (Phaseolus vulgaris L.). Acta Agriculturae Slovenica, 83: 181-190.
Mosahebi, G. H. 1972. Study on Bean common mosaic virus and their-properties in Iran. M.Sc. Thesis, University of Tehran. pp 185. (In Persian).
Mostafavi, F. S. 2014. Serological and molecular detection of Bean common mosaic virus in Gorgan area. MSc. Thesis, Gorgan University of Agricultural Science and Natural Resources. pp 125. (In Persian).
Mousavi, S. M. 2014. Determination of some serological and molecular characteristics of BCMV isolates in Khorasan Razavi Province. M.Sc. Thesis, Alzahra University. pp 101. (In Persian).
Naderpour, M. 1999. Identification of pathotypes and strains of Bean common mosaic virus in Tehran province. M.Sc. Thesis, University of Tehran. pp 112. (In Persian).
Naderpour, M., Lund, O. S. and Johansen, I. E. 2009. Sequence analysis of expressed cDNA of Bean common mosaic virus RU1 isolate. Iranian Journal of Virology, 3(3): 39-41.
Najafi, G. H. 1969. Race of Bean mosaic virus in Iran and studies about it. Journal of Plant Diseases, 5: 124-133.
Pasev, G., Kostova D. and Sofkova, S. 2013. Identification of genes for resistance to Bean common mosaic virus and Bean common mosaic necrosis virus in snap bean (Phaseolus vulgaris L.) breeding lines using conventional and molecular methods. Journal of Phytopathology, 162: 19-25.
Peyambari, M., Koohi Hbibi, M., Mosahebi, G. H. and Izadpanah, K. 2011. Study on Bean common mosaic virus (BCMV) in some provinces and the reaction of bean genotypes to this virus. Journal of Plant Protection (Agricultural Science and Technology), 25: 250-257.
Salari, N., Seied mousavi, M., Shahraeen, N., Ghorbani, S. and Maleki, M. 2013. Detection and identification of Bean common mosaic virus and Bean common mosaic necrosis virus using Immunocapture RT-PCR. Cellular and Molecular Biotechnology News, 11: 21-28. (In Persian).
Saqib, M., Jones, R. A. C., Cayford, B. and Jones, M. G. K. 2005. First report of Bean common mosaic virus in western Australia. Plant Pathology, 54: 563.
Saqib, M., Nouri, S., Cayford, B., Jones, R. A. C. and Jones, M. G. K. 2010. Genome sequences and phylogenetic placement of two isolates of Bean common mosaic virus from Macroptilium atropurpureum in north-west Australia. Australasian Plant Pathology, 39(2): 184-191.
Sayyahi, C. 2010. Study on serological and molecular characteristics of BCMV isolates in Fars province. MSc. Thesis. Azad University, Science and Research Branch, Tehran. pp 222. (In Persian). 
Shahraeen, N., Hasani Mehrban, A., Pourdavaei, H., Beizaei, A., Mostaed, M. and Bananej, K. 2001. Determining the sources of genetic resistance to three important viruses of beans CMV, BYMV and BCMV. Khomein station and Boroujerd research project.
Silbernagel, M. J., Mink, G. I., Zhao, R. L. and Zheng, G. Y. 2001. Phenotypic recombination between Bean common mosaic and Bean common mosaic necrosis potyviruses in vivo. Archives of Virology, 146: 1007-1020.
Stewart, V. B. and Reddick, D. 1917. Bean mosaic. Journal of Phytopathology-Phytopathologische Zeitschrift, 7: 61-72.
Strausbaugh, C. A., Miklas, P. N., Singh, S. P., Myers, J. R. and Forster, R. L. 2003. Genetic characterization of differential reactions among host group 3 common bean cultivars to NL-3 K strein of Bean common mosaic necrosis virus. Journal of Phytopathology-Phytopathologische Zeitschrift, 93: 683-690.
Zhou, G. C., Wu, X. Y., Zhang, Y. M., Wu, P., Wu, X. Z., Liu, L.W., Wang, Q., Hang, Y. Y., Yang, J. Y., Shao, Z. Q., Wang, B. and Chen, J. Q. 2014. A genomic survey of thirty soybean-infecting Bean common mosaic virus (BCMV) isolates from China pointed BCMV as a potential threat to soybean production. Virus Research, 191: 125-133.