شناسایی و بررسی مولکولی ویروس موزاییک معمولی لوبیا (BCMV) و موزاییک نکروتیک معمولی لوبیا (BCMNV) در استان مازندران

پژوهش های حفاظت گیاهان ایران

مقاله 16، دوره 30، شماره 1، خرداد 1395، صفحه 143-150   

نوع مقاله: مقالات پژوهشی

شناسه دیجیتال (DOI): 10.22067/jpp.v30i1.41631

نویسندگان

زهره مرادی¹؛ محسن مهرور^{* 1}؛ احسان نظیفی²

¹دانشگاه فردوسی مشهد

²دانشگاه مازندران

چکیده

طی سال های 1392 و 1393، تعداد 50 نمونه برگ لوبیا دارای علائم از مزارع استان مازندران جمع آوری و RNA کل آنها استخراج شد. طی آزمون RT-PCR با استفاده از یک جفت آغازگر دژنره منطبق بر ژن کدکننده پروتئین Cylindrical inclusion (CI) قطعه ای به طول bp680 در 13 نمونه تکثیر گردید. تکثیر اختصاصی نمونه های آلوده با آغازگرهای CIF/R، نشان از آلودگی به یک پوتی ویروس داشت. محصول PCR مربوط به دو جدایه پس از همسانه سازی در حاملpTG19-T ، توالی یابی شد. آنالیز بلاست توالی ها نشان داد که قطعات تکثیر شده در واکنش PCR متعلق به ویروس موزاییک نکروتیک معمولی لوبیا (Bean common mosaic necrosis virus, BCMNV) (از لوبیا سفید) و ویروس موزاییک معمولی لوبیا (Bean common mosaic virus, BCMV) (از لوبیا چشم بلبلی) است. آنالیز فیلوژنتیکی جدایه های تعیین ترادف شده در ناحیه ژنی CI نشان داد که جدایه ی تعیین توالی شده BCMV-MAZ (از ساری) در سطح نوکلئوتیدی و اسید آمینه ای به ترتیب دارای بیشترین شباهت (8/96% و 7/98%) با جدایه آمریکا (RU1M) و کمترین شباهت (5/79% و 6/91 %) با جدایه کره جنوبی (Habin1) می باشد. همچنین جدایه BCMNV-MAZ (از جویبار) بیشترین شباهت (8/97%) را با جدایه های آمریکایی NL-3 K،NL5 و کمترین شباهت (9/96%) را با NL8 در سطح نوکلئوتیدی دارد. در سطح اسید آمینه ای نیز 7/97 % شباهت با سایر جدایه های مورد بررسی دارد. این اولین گزارش از وجود ویروس های مذکور در استان مازندران به روش مولکولی است.

کلیدواژه‌ها

BCMV؛ BCMNV؛ RT-PCR؛ آنالیز فیلوژنتیک

مراجع

1- Adams M.J., Antoniw J.F., and Fauquet C.M. 2005. Molecular criteria for genus and species discrimination within the family Potyviridae. Archives of Virology, 150:459–479.

2- Baradaran G.R., and Jafarpour B. 1998. Study on occurrence of bean common mosaic virus and Cucumber mosaic virus in Bean fields of Mashhad and Chenaran area. p. 157-163. Proceedings of the 13th Iranian plant protection congress, 23-27 Aug. 1998. Karaj, Iran. (In Persian)

3- Carrington J.C., Jensen P.E., and Schaad M.C. 1998. Genetic evidence for an essential role for potyvirus CI protein in cell-to-cell movement. Plant Journal, 14(4):393-400.

4- Chen J., and Adams M.J. 2001. A universal PCR primer to detect members of the Potyviridae and its use to examine the taxonomic status of several members of the family. Archives of Virology, 146:757-766.

5- Dorri H., lak M., bani-jamali S.M., Dadivar M., Ghanbari A., Khodshenas M., and Asadi B. 2003. Beans (from planting to harvest). Journal of Extension Education, 305, 77 p.(In Persian)

6- Drijfhout E. 1991. Bean common mosaic. Compendium of Bean Diseases. APS Press, pp 37-39.

7- Flores-Estevez N., Acosta-Gallegos J.A., Silva-Rosales L. 2003. Bean common mosaic virus and Bean common mosaic necrosis virus in Mexico. Plant Disease, 87:21-25.

8- Gibbs A.J., Mackenzie A.M., and Gibbs M.J. 2003. The 'potyvirid primers' will probably provide phylogenetically informative DNA fragments from all species of Potyviridae. Journal of Virological Methods, 112:41-44.

9- Ha C., Coombs S., Revill P.A., Harding R.M., Vu M., and 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, 153(1):25-36.

10- Hsu Y.C., Yeh T.J., and Chang Y.C. 2005. A new combination of RT-PCR and reverse dot blot hybridization for rapid detection and identification of potyviruses. Journal of Virological Methods, 128:54-60.

11- Hull R. 2014. Plant virology (5th ed.). New York: Academic Press.1098 p.

12- Kelly J.D. 1997. A review of varietal response to bean common mosaic potyvirus in Phaseolus vulgaris. Plant Varieties and Seed, 10:1-6.

13- Khan J.A., LohuisD., Goldbach R., and Dijkstra J. 1993. Sequence data to settle the taxonomic position of Bean common mosaic virus and Blackeye cowpea mosaic virus isolates. Journal of General Virology, 74: 2243-2249.

14- Klein P.G., Klein R.R., Rodriguez-Cerezo E., Hunt A.G., and Shaw J.G. 1994. Mutational analysis of the Tobacco vein mottling virus genome. Virology, 204:759-769.

15- Lopez-Moya J.J., Valli A., and Garcıa J.A. 2009. Potyviridae. Encyclopedia of Life Sciences. Annual Review of Phytopathology, 34:227–247.

16- Lee K.C., Wong S.M., Mahtani P.H., and Chng C.G. 1997. Sequence and phylogenetic analysis of the cytoplasmic inclusion protein gene of Zucchini yellow mosaic potyvirus: its role in classification of the Potyviridae. Virus Genes, 14:41–53.

17- Mavric I. and Sustar-Vozlic J. 2004. Virus diseases and resistance to Bean commonmosaic and Bean common mosaic necrosis potyvirus in common bean (Phaseolusvulgaris L.). Acta agriculturae slovenica, 83:181-190.

18- McKern N.M., Mink G.I., BarnettO.W., MishraA., WhittakerL.A., SilbernagelM.J., Ward C.W. and Shukla D.D. 1992a. Isolates of Bean common mosaic virus comprising two distinct potyviruses. Phytopathology, 82:923-929.

19- McKern, N. M., Ward C.W., and Shukla D.D. 1992b. Strains of Bean common mosaic virus consist of at least two distinct potyviruses. Archives of Virology, [Suppl.5]:407-414.

20- Mink G.I., and Silbernagel M.J. 1992. Serological and biological relationship among viruses in the Bean common mosaic virus subgroup. In: Barnett OW (ed) Potyvirus taxonomy. Springer, Wien New York. Archives of Virology, pp 397-406.

21- Naderpour M.,Mosahebi G., and Kohi Habibi M. 2000. III, Vta & Vib Pathotypes and NL-5, NL-8 & NL-3 necrotic strains of BCMV, a new record for Iran. P. 280-283. Proceedings of the 14th Iranian plant protection congress, 5-8 Sep. 2000. Isfahan, Iran. (In Persian)

22- Naderpour M., Mosahebi G., and Kohi Habibi M. 2000. Detection of BCMV serotypes by using of physicochemical methods. p. 284-287. Proceedings of the 14th Iranian plant protection congress, 5-8 Sep. 2000. Isfahan, Iran. (In Persian)

23- Peyambari, M., Kohi habibi M., Mosahebi G., and Izatpanah K. 2011. Consideration of BCMV in some provinces of Iran and three genotypes response to BCMV. Journal of Plant Protection, 25(3):250-257.

24- Pudashini B.J., Shahid M.S., and Natsuaki K.T. 2013. First report of Bean common mosaic necrosis virus (BCMNV) infecting sweet bean in Nepal. Plant Disease, Vol. 97 No. 2 pp. 290.

25- Revers F., Le Gall O., Candresse T., and Maule A.J. 1999. New advances in understanding the molecular biology of plant: potyvirus interactions. Molecular Plant-Microbe Interactions, 12:367-376.

26- Saiz M., de Blas C., Carazo G., Fresno J., Romero J., and Castro S. 1995. Incidence and characterization of Bean common mosaic virus isolates in Spanish bean fields. Plant disease, 79:79-81.

27- Salari N., Seyed Musavi M., Shahraeen N., Ghobani S., and Maleki M. 2013. Identification and diagnosis of isolates of Bean common mosaic virus and bean common mosaic necrosis virus by immunocapture RT-PCR. New Cellular and Molecular Biotechnology Journal, 3 (11):21-28. (In Persian)

28- Shahraein N., Hasani mehraban H., Poordavaei H., Beyzaei A., Mostaed M., and Bananj k. 2001. Determination of genetically sources of resistance to three importance common bean viruses, CMV, BYMV and BCMV. Research final report, Khomain and bojnoord station (In Persian).

29- Shahraein N., Hasani mehraban H., Mostaed M., and Ghotbi T. 2002. Report on Bean common mosaic necrosis virus infecting French bean in Arak and Lorestan provinces.p. 282-287. Proceedings of the 15th Iranian plant protection congress, 7-11 Sep. 2002. Kermanshah, Iran. (In Persian)

30- Shukla D.D., Ward C.W., and Brunt A.A. 1994. The Potyviridae. CAB International, Walling ford, UK. p. 74-112.

31- 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. Archive of Virology, 146:1007-1020.

32- Strausbaugh C.A., Miklas P.N., Singh S.P., Myers J.R., and Forster R.L. 2003. Genetic characterization of differential reaction among host group 3 common bean cultivars to NL-3 k strain of Bean common mosaic necrosis virus. Phytopathology, 93:683-690.

33- Tamura K., Peterson D., Peterson N., Stecher G., Nei M., and Kumar S. 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28:2731-2739.

34- Vallejos C.E., Gustavo A.M., Valerie J., Tommy R.P., Ney S.S, and Sally A.M. 2006. Genetic and molecular characterization of the I locus of Phaseolus vulgaris. Genetics, 172:1229-1242.

35- Vetten H.J., Lesemann D.E., and Maiss E. 1992. Serotype A and B Strain of Bean common mosaic virus are two distinct Potyvirus. Archive of Virology, Suppl 5:415-431.

36- Yazdi Samadi B., and Abdemeyshaei S. 1996. Agronomical plant breeding. Iran University press. PP 283 (In Persian)

37- Zheng H., Chen J., Chen J., Adams M.J., and Hou M. 2002. Bean common mosaic virus isolates causing different symptoms in asparagus bean in China differ greatly in the 5' parts of their genomes. Archives of Virology, 147:1257–1262.