c3518cb17d976b8

مقایسه تنوع ژنتیکی ناحیه 3′UTR جدایه‌های ویروس سوختگی سیاه برگ چغندرقند (Beet black scorch virus) خراسان با جدایه‌های ایران و سایر نقاط دنیا

نوع مقاله : مقاله پژوهشی

نویسندگان

1 مربی پژوهشی، بخش تحقیقات چغندرقند، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان خراسان رضوی، سازمان تحقیقات، آموزش و ترویج

2 دانشیار ،گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه فردوسی مشهد،ایران

3 دانشیار، مؤسسه تحقیقات اصلاح و تهیه بذر چغندرقند، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

ویروس سوختگی سیاه برگ چغندرقند (Beet black scorch virus, BBSV) ازجمله ویروس‌های خاک‌برد چغندرقند است. در این تحقیق، 80 نمونه چغندرقند از مزارع خراسان رضوی و شمالی جمع‌آوری شد. پس از استخراج RNAی کل آن‌ها، ناحیه 3'UTR ژنوم ویروس به تعداد 315 نوکلئوتید با استفاده از واکنش زنجیره‌ای پلیمراز با رونویسی معکوس و بهره‌گیری از آغازگرهای اختصاصی رفت و برگشتی تکثیر شد. این ناحیه در شش جدایه مختلف پس از همسانه سازی، توالی‌یابی و در NCBI ثبت شدند. نتایج بررسی خویشاوندی این جدایه‌ها (بر اساس (3'UTR با کل جدایه‌های ثبت‌شده در بانک جهانی نشان داد که آن‌ها در دو گروه عمده I و II قرار می‌گیرند به‌طوری‌که هر یک به دو زیرگروه تقسیم می‌شوند. شش جدایه این بررسی با 23 جدایه از نقاط مختلف ایران در زیرگروه IA و چهار جدایه دیگر از خراسان در زیرگروه IB جای گرفتند. سه جدایه از کرمانشاه و یک جدایه از همدان (Ir-KSh5,Ir-KSh4 Ir-KSh6 و Ir-Ha2) در زیرگروه IIA قرار گرفتند. چهار جدایه از کرمانشاه Ir-KSh9,Ir-KSh8,Ir-KSh7) و Ir-KSh10) به همراه جدایه‌های چینی، اروپایی و آمریکایی زیرگروه IIB را تشکیل دادند. نتایج مقایسه نوکلئوتیدی توالی شش جدایه نشان داد که اگرچه تغییر نوکلئوتیدی مهم در موقعیت 3477 (یوریدین بجای گوانین) صورت نگرفته است، اما در موقعیت 3398 آدنین جایگزین گوانین و در موقعیت 3393 فقط در جدایه فریمان (MW274750) یوریدین بجای سیتوزین جایگزین شده است. نتایج مقایسه میانگین شباهت نوکلئوتیدی ناحیه 3'UTR این جدایه‌ها نشان داد که با جدایه‌های ایرانی (به غیر از جدایه-های کرمانشاه با 52/92 درصد) در دامنه بین 75/98-37/97 درصد مشابهت دارند.

کلیدواژه‌ها


عنوان مقاله [English]

Comparison of genetic variation of 3′UTR region of Beet black scorch virus in Khorasan isolates with the other isolates from Iran and the world

نویسندگان [English]

  • Jamshid soltani Idliki 1
  • mohsen mehrvar 2
  • seyed bagher mahmoudi 3
1 Sugar Beet Research Department, Khorasan Razavi Agricultural and Natural Resources Research and Education Center, AREEO, Mashhad, Iran.
2 Associate Professor, Department of Plant Pathology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
3 Associate Professor, Sugar Beet Seed Institute, Agricultural Research, Education, and Extension Organization (AREEO), Karadj, Iran
چکیده [English]

Beet black scorch virus is one of the soil-borne sugar beet viruses in Iran. In this research work, we collected 80 samples of sugar beet farms in Razavi and Northern Khorasan Provinces. After extraction of their total RNA, the 3'UTR region of the virus genome was amplified to 315 nucleotides using reverse transcription polymerase chain reaction using specific primers. The 3'UTR of six different isolates was sequenced following cloning and then submitted to NCBI. Phylogenetic analysis based on nucleotide sequences of the 3'UTR region showed that the entire world isolates could be classified into two main groups II and I and each one was divided into two subgroups. Isolates of this study were grouped with 23 isolates from Iran in subgroup IA and Four isolates from Khorasan province were placed in the IB subgroup. Four Iranian BBSV isolates from the West were grouped in the subgroup GIIA. All Chinese isolates and the European isolates together with the USA isolate clustered in subgroup GIIB. The results of nucleotide comparison of six isolates showed that although no mutation occurred at position 3477 (uridine instead of guanine), However, in position 3398, adenine replaced guanine (similar to 60% of Iranian isolates) and in position 3393, uridine was substituted for cytosine only in Fariman isolates (MW274750). The results of comparing the average nucleotide identity of 3'UTR isolates in this study showed that they are identical to Iranian isolates in the range from 97.75% to 97.98%. Nevertheless, they had the least identity with Kermanshah isolates (92.52%).

کلیدواژه‌ها [English]

  • "Plant viruses"
  • "sugar beet diseases"
  • "sequencing"
  1. Cai, Z. N. Chen, D. H. Wu, M. S. Cui, X. M. Yu, J. L., & Liu, Y. (1993).Identification of pathogenic virus of Beet black scorch virus disease and detection by synthesized cDNA probes. Beijing Agric. Univ, 19, 112.
  2. Cao, Y. Cai, Z. Ding, Q. Li, D. Han, C. Yu, J., & Liu, Y. (2002). The complete nucleotide sequence of Beet black scorch virus (BBSV), a new member of the genus Necrovirus. Archives of Virology, 147(12), 2431–2435.
  3. Cui, X. (1988). An icosahedral virus found in sugar beet. J Xinjiang Shihezi Agric College, 10, 73–78.
  4. Cui, X. M. Cai, Z. N. Wu, J., and Liu, Y. (1991). Study on symptom pattern of sugarbeet rhizomania disease. Plant Protect, 17, 5–7.
  5. Farzadfar, S., & Pourrahim, R. (2019). Positive selection and recombination shaped the large genetic differentiation of Beet black scorch virus PLoS ONE, 14(4), 1–19.
  6. Gonzalez-Vazquez, M. Ayala, J. Garcia-Arenal, F., & Fraile, A. (2009). The occurrence of Beet black scorch virus infecting sugar beet in Europe. Plant Disease, 93(1), 21–24.
  7. Guo, L.H. Cao, Y.H. Li, D.W. Niu, S.N. Cai, Z.N. Han, C.G. Yu, J.L. (2005). Analysis of nucleotide sequences and multimeric forms of a novel satellite RNA associated with Beet black scorch virus. Journal of Virology, 79(6), 3664–3674.
  8. Hull, R. (2002). Virus Infection, Plant. In Encyclopedia of Molecular Biology. Hoboken, NJ, USA: John Wiley and Sons, Inc.
  9. Junxi, J. Jingfeng, Z. Shaochen, C. Dajin, Y. Jialin, Y. Zhunan, C., & Yi, L. (1999). Transmission of Beet black scorch virus by Olpidium brassicae. Acta Agriculturae Universitatis Jiangxiensis, 21(4), 525-528.
  10. Koenig, R., and Valizadeh, J. (2008). Molecular and serological characterization of an Iranian isolate of Beet black scorch virus. Archives of Virology, 153(7), 1397–1400.
  11. Kumar, S. Stecher, G. Li, M. Knyaz, C., & Tamura, K. (2018). MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35(6), 1547–1549.
  12. Li, J. Li, M. Li, C. Gao, Y. Li, D. Han, C., & Yu, J. (2008). Effects on the local symptoms of subgenomic RNAs expressions and their translational products of Tobacco necrosis virus A Chinese isolate. Chin Sci Bull, 53.
  13. Lommel, S. A. (2005). Family Tombusviridae. Virus Taxonomy. Eighth Report of the International Committee on Taxonomy of Viruses, San Diego, USA, 2005.
  14. Mehrvar, M., & Bragard, C. (2011). Iranian diversity of Beet black scorch virus and satellite. In Proceedings of the Eighth Symposium of the International Working Group on Plant Viruses with Fungal Vectors, Louvain-La-Neuve, Belgium, 6-8 July 2011 (pp. 41–46). International Working Group on Plant Viruses with Fungal Vectors.
  15. Mehrvar, M. Valizadeh, J. Koenig, R., & Bragard, C. G. (2009). Iranian Beet necrotic yellow vein virus (BNYVV): pronounced diversity of the p25 coding region in A-type BNYVV and identification of P-type BNYVV lacking a fifth RNA species. Archives of Virology, 154(3), 501–506.
  16. Merhvar, M., & Bragard, C. (2009). Beet black scorch virus in Iran is more diverse than anywhere. Phytopathology, 99(6), S84
  17. Meunier, A. Schmit, J. F. Stas, A. Kutluk, N., & Bragard, C. (2003). Multiplex reverse transcription-PCR for simultaneous detection of Beet necrotic yellow vein virus, Beet soilborne virus, and Beet virus Q and their vector Polymyxa betae KESKIN on sugar beet. Applied and Environmental Microbiology, 69(4), 2356–2360.
  18. Muhire, B. M. Varsani, A., & Martin, D. P. (2014). SDT: a virus classification tool based on pairwise sequence alignment and identity calculation. PloS One, 9(9).
  19. Ratti, C. Clover, G. R. G. Autonell, C. R. Harju, V. A., and Henry, C. M. (2005). A multiplex RT-PCR assay capable of distinguishing Beet necrotic yellow vein virus types A and B. Journal of Virological Methods, 124(1–2), 41–47.
  20. Samiei, A. Mehrvar, M., & Zakiaghl, M. (2017). Genetic diversity and distribution of Beet black scorch virus in some provinces of Iran. Journal of Plant Protection. 20(4), 540–547.
  21. Shen, R., & Miller, W. A. (2007).Structures required for poly (A) tail-independent translation overlap with but are distinct from, cap-independent translation and RNA replication signals at the 3′ end of Tobacco necrosis virus Virology, 358(2), 448–458.
  22. Weiland, J J. Larson, R. L. Freeman, T. P., & Edwards, M. C. (2006). First report of Beet black scorch virus in the United States. Plant Dis, 90.
  23. Weiland, John J, Van Winkle, D. Edwards, M. C. Larson, R. L. Shelver, W. L. Freeman, T. P., & Liu, H.Y. (2007). Characterization of a U.S. Isolate of Beet black scorch virus. Phytopathology, 97(10), 1245–1254.
  24. Xu, J. Liu, D. Zhang, Y. Wang, Y. Han, C. Li, D., & Wang, X.B. (2016). Improved Pathogenicity of a Beet black scorch virus Variant by Low Temperature and Co-infection with Its Satellite RNA. Frontiers in Microbiology, 7, 1771.
  25. Xu, J. Wang, X. Shi, L. Zhou, Y. Li, D. Han, C., & Yu, J. (2012). Two distinct sites are essential for virulent infection and support of variant satellite RNA replication in spontaneous Beet black scorch virus Journal of General Virology, 93(12), 2718–2728.
  26. Zhang, X. Zhao, X. Zhang, Y. Y. Niu, S. Qu, F. Zhang, Y. Y., & Li, D. (2013). N-terminal basic amino acid residues of Beet black scorch virus capsid protein play a critical role in virion assembly and systemic movement. Virology Journal, 10(1), 200.
  27. Zhang, Y. Zhang, X. Niu, S. Han, C. Yu, J., & Li, D. (2011). Nuclear localization of Beet black scorch virus capsid protein and its interaction with importin α. Virus Res, 155(1), 307–315.