c3518cb17d976b8

بررسی سه جدایۀ ویروس وای سیب‌زمینی جداشده از استان خوزستان بر پایۀ ژن پروتئین پوششی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق کارشناسی ارشد، دانشکدۀ کشاورزی دانشگاه ولیعصر(عج) رفسنجان

2 استادیار، دانشکدۀ کشاورزی دانشگاه ولیعصر(عج) رفسنجان

چکیده

ویروس وای سیب‌زمینی (Potato virus Y, PVY) یکی از ویروس­های مهم زیانبار با دامنۀ میزبانی گسترده است. در این پژوهش با توجه به شناسایی ویروس وای سیب‌زمینی و جداسازی آن از کشتزارهای شلغم استان خوزستان در سال زراعی 1392، ویژگی­های سه جدایۀ انتخابی از شهرستان­های دزفول، شوشتر و شوش (7Tu، 9Tu و 10Tu) بررسی شد. در آزمون تعیین دامنۀ میزبانی، سه جدایه روی گیاهانی متعلق به سه خانوادۀ Solanaceae، Chenopodiaceae و Amaranthaceae، به‌صورت مکانیکی مایه­زنی شدند. هر سه جدایه روی سیب‌زمینی، گوجه‌فرنگی، فلفل، تاتوره و گل تکمه‌ای نشانه‌های موضعی و فراگیر (سیستمیک) ایجاد کردند ولی روی بوته‌های سلمه‌تره، نشانه‌هایی مشاهده نشد. در آزمون SDS-PAGE، دو باند با اندازۀ حدود 33 و 31 کیلودالتونی در آمودۀ خالص‌شدۀ هر سه جدایۀ خالص‌سازی‌شده مشاهده شد. برای تعیین سویۀ جدایه‌های انتخابی از جفت آغازگرهای اختصاصی سویۀ PVY (CF/OR, NF/NR, OF/OR و PVY3TIM/PVNIBIP) که بر پایۀ ناحیۀ (CP) طراحی شده‌اند استفاده شد. در آزمون PCR با آغازگرهای اختصاصی سویۀ O (OR/OF) و سویۀ NTN (PVY3TIM وPVNIBIP) به ترتیب دو قطعۀ 608 و 1200 جفت بازی در هر سه جدایه افزایش شد. در ادامه، ناحیۀ CP هر سه جدایه‌، افزایش، همسانه‌سازی و توالی­یابی شد. درنهایت با استفاده از مجموع نتایج به‌دست‌آمده از آزمون­های زیستی (بیولوژیکی) و بررسی توالی نوکلئوتیدی و آمینواسیدی مشخص شد که جدایه‌های 7Tu، 9Tu و 10Tu جداشده از کشتزارهای شلغم استان خوزستان بیشترین همسانی را با سویه‌های NTN و W ویروس PVY دارند. این نخستین گزارش از شناسایی ویروس وای سیب‌زمینی در کشتزارهای شلغم است.

کلیدواژه‌ها


عنوان مقاله [English]

The characterization of three isolates of Potato virus Y from khuzestan province according to the coat protein

نویسندگان [English]

  • Seyed Sajjad Zabihi 1
  • Samin-o-Sadat Hosseini 2
  • Seyed Ahmad Hosseini 2
1 Former M. Sc. Student, Faculty of Agricluture, Vali-e-Asr University of Rafsanjan, Iran
2 Assistant Professor, Faculty of Agricluture, Vali-e-Asr University of Rafsanjan, Iran
چکیده [English]

Potato virus Y is one of the most important damaging viruses with a broad host range. In this research due to detection and isolation of Potato virus Y in turnip fields of Khuzestan province during 2014 growing season, characteristics of three selected isolates (7Tu, 9Tu and 10Tu) from Dezful, Shushtar and Shush cities were investigated. To analyze the host range of the isolates, three selected isolates were mechanically inoculated on some members of Solanaceae, Chenopodiaceae, and Amaranthaceae. All three isolates induced local and systemic symptoms on the leaves of potato, tomato, pepper, datura, and gomphrena plants, but no symptoms were observed on Chenopodium quinoa. Analysis of the PVY partially purified preparations indicated that molecular weight of two protein bands obtained by SDS-PAGE was estimated to be about 33 and 31 KDa. To determine the strain of selective isolates the specific primer pairs CF/OR, NF/NR, OF/OR, and NF/OR were used. Using OF/OR and PVY3TIM/PVNIBIP primers, two fragments with 609 and 1200 bp length respectively were amplified. The CP region in the all three isolates was amplified, cloned and sequenced. The results of all aforementioned tests showed that 7Tu, 9Tu, and 10Tu isolates are probably a member of NTN or W strain groups. According to our knowledge, this is the first report on occurrence of PVY in turnip fields.

کلیدواژه‌ها [English]

  • Coat protein
  • turnip
  • Potato virus Y
  • RT-PCR
  • SDS-PAGE
Amer, M., El-Hammady, M., Mazyad, H., Shalaby, A. & Abo-Elabbas, F. (2004). Cloning, expression and nucleotide sequence of Coat protein gene of an Egyptian isolate of Potato Virus Y strain NTN infecting Potato plants. Egyptian Journal Virology, 1, 39-50.
Bimboim, H. C. & Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic acids Research, 7(6), 1513-1523.‏
Boonham, N., Walsh, K., Hims, M., Preston, S., North, J. & Barker, I. (2002). Biological and sequence comparisons of Potato virus Y isolates associated with Potato tuber necrotic ringspot disease. Plant Pathology, 51, 117-126.
Clark, M. F. & Adams, A. (1977). Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology, 34, 475-483.
Fakhrabad, F. Z., Nasrollanejad, S., Ahmadikhah, A. & Taghinasab, M. (2012). Sequencing of three isolates and prevalence of Potato virus Y in Tobacco fields of Golestan province, and phylogenetic comparison of the Iranian and world isolates of the virus. Iranian Journal of Plant Pathology, 48.
Folwarczna, J., Plchova, H., Moravec, T., Hoffmeisterova, H., Dedic, N. & Cerovska, N. (2008). Production of polyclonal antibodies to a recombinant Coat protein of Potato virus Y. Folia Microbiologica, 53(5), 438-442.
Fuji, S., Inoue, M., Yamamoto, H., Furuya, H., Naito, H. & Matsumoto, T. (2003). Nucleotide sequences of the Coat protein gene of Potyviruses infecting Ornithogalum thyrsoides. Archives of Virology, 148, 613-621.
Ha, Cuong Viet. (2007). Detection and identification of potyviruses and geminiviruses in Vietnam. Ph.D.  thesis. by Publication, Queensland University of Technology, Queensland.
Higgins, C., Cassidy, B., Teycheney, P.-Y., Wongkaew, S. & Dietzgen, R. (1998). Sequences of the Coat protein gene of five Peanut stripe virus (PStV) strains from Thailand and their evolutionary relationship with other Bean common mosaic virus sequences. Archives of Virology, 143, 1655-1667.
Hosseini, S., Koohi Habibi, M., Mosahebi, G., Motamedi, M. & Winter, S. (2012). First report on the occurrence of Tobacco streak virus in Sunflower in Iran. Journal of Plant Pathology, 94, 585-589.
Hull, R. (2002). Induction of disease 1: virus movement through the plant and effects on plant metabolism. Matthew’s plant Virology, 4th edn, Academic Press, San Diego, 373-436.
Huttinga, H. & Mosch, W. (1974). Properties of viruses of the Potyvirus group. 2. Buoyant density, S value, particle morphology, and molecular weight of the Coat protein subunit of Bean yellow mosaic virus, Pea mosaic virus, Lettuce mosaic virus, and Potato virus YN. Netherlands Journal of Plant Pathology, 80, 19-27.
Inoue-Nagata, A. K., Fonseca, M. e. N., Lobo, T. O., Ávila, A. C. & Monte, D. C. (2001). Analysis of the nucleotide sequence of the Coat protein and 3'-untranslated region of two Brazilian Potato virus Y isolates. Fitopatologia Brasileira, 26, 45-52.
Jensen, S. G., Long-Davidson, B. & Seip, L. (1986). Size variation among proteins induced by Sugarcane mosaic viruses in plant tissue. Journal Series Paper, 68583, 0722.
Kachroo, P., Yoshioka, K., Shah, J., Dooner, H. K. & Klessig, D. F. (2000). Resistance to Turnip crinkle virus in Arabidopsis is regulated by two host genes and is salicylic acid dependent but NPR1, ethylene, and jasmonate independent. The Plant Cell, 12, 677-690.
Kuchacki, A. R., Nasirimahalati, M., Hassanzade Aval, F., Mansuri, H., Amiri, S. R., Zarghani, H. & Karimian, M. (2013). Assessment of biodiversity vegetables in ecosystems crop of Iran. Journal of Applied Ecology, 4(2), 1-11. (in Farsi)
Khuzestan Agriculture Jahad. (2010). Turnip harvest from the fields of Khuzestan province. Retrieved December 22, 2014, from http://www.kaj.maj.ir/ravabetomome/news.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
Leisner, S. M., Turgeon, R. & Howell, S. H. (1992). Long distance movement of Cauliflower mosaic virus in infected Turnip plants. Molecular. Plant-Microbe Interact, 5, 41-47.
Missiou, A., Kalantidis, K., Boutla, A., Tzortzakaki, S., Tabler, M. & Tsagris, M. (2004). Generation of transgenic Potato plants highly resistant to Potato virus Y (PVY) through RNA silencing. Molecular Breeding, 14, 185-197.
Moghal, S. & Francki, R. (1976). Towards a system for the identification and classification of Potyviruses: I. Serology and amino acid composition of six distinct viruses. Virology, 73, 350-362.
Moury, B., Palloix, A., Caranta, C., Gognalons, P., Souche, S., Selassie, K. G. & Marchoux, G. (2005). Serological, molecular, and pathotype diversity of Pepper veinal mottle virus and Chili veinal mottle virus. Phytopathology, 95, 227-232.
Nanayakkara, U. N., Singh, M., Pelletier, Y. & Nie, X. (2012). Investigation of Potato virus Y (PVY) strain status and variant population in Potatoes in New Brunswick, Canada. American Journal of Potato Research, 89, 232-239.
Ohshima, K., Sako, K., Hiraishi, C., Nakagawa, A., Matsuo, K., Ogawa, T., Shikata, E. & Sako, N. (2000). Potato tuber necrotic ringspot disease occurring in Japan: its association with Potato virus Y necrotic strain. Plant Disease, 84, 1109-1115.
Sabir, J. S. (2012). Identification of six Potato virus Y isolates from Saudi Arabia. African Journal of Biotechnology, 11, 9709-9715.
Singh, R. P., Valkonen, J. P., Gray, S. M., Boonham, N., Jones, R., Kerlan, C. & Schubert, J. (2008). Discussion paper: The naming of Potato virus Y strains infecting Potato. Archives of Virology, 153, 1-13.
Smith, K. M. (1931). Composite nature of certain Potato viruses of the mosaic group. Nature, 127, 702.
Urcuqui-Inchima, S., Haenni, A.-L. & Bernardi, F. (2001). Potyvirus proteins: a wealth of functions. Virus Research, 74, 157-175.
Visser, J. C. (2012). A study of the strain evolution and recombination of South African isolates of Potato virus Y. Ph. D. thesis .Stellenbosch University, Stellenbosch.