شناسایی ویروس رگبرگ روشنی زرد مرکبات در استان مازندران

نوع مقاله: علمی پژوهشی-فارسی

نویسندگان

1 استادیار، پژوهشکده مرکبات و میوه‌های نیمه‌گرمسیری، رامسر

2 دانشیار، پژوهشکده مرکبات و میوه‌های نیمه‌گرمسیری، رامسر

10.22055/ppr.2020.15997

چکیده

ویروس رگبرگ روشنی زرد مرکبات (Citrus yellow vein clearing virus)، ویروسی خسارت‌زا از جنس Mandarivirus است که در سال‌های اخیر وقوع آن پس از کشورهای پاکستان، هند، ترکیه و چین، از ایران هم گزارش شده است. در میزبان‌های حساس شناخته شده در کشور شامل نارنج، لمون و پرشین‌لایم، علائم زردی رگبرگ‌های جانبی با نواحی آب‌سوخته متناظر در پشت برگ ظاهر می‌شود. این علائم مبنای تشخیص اولیه بیماری در نهال‌ها و درختان نارنج‌، لمون و پرشین‌لایم در نهالستان‌ها و باغ‎های مرکبات استان مازندران قرار گرفت. از تعداد 53 نمونه جمع‌آوری شده از شهرستان‌های این استان در بهار سال‌های 1395، 1396 و 1397، سه جدایه SOU، LEN و LIE از میزبان‌های فوق جهت آزمون‌های تکمیلی انتخاب شدند. انتقال مکانیکی و پیوندی این جدایه‌ها در گلخانه با دمای کنترل شده به گیاهان لوبیا، نارنج و لمون با موفقیت انجام شد. در شرایط باغ نیز، علائم شاخص بیماری پس از پیوند ارقام حساس عاری از ویروس روی تنه درختان بیمار به دست آمد. به منظور تأیید آلودگی گیاهان محک نارنج و لمون و نمونه‌های باغی، استخراج آران­ا کل به روش SDS-Potassium acetate از بافت‌های جوان علائم‌دار انجام پذیرفت. بررسی نمونه­ ها با استفاده از آزمون RT-PCR دو مرحله‌ای و جفت آغازگر‌ اختصاصی ژن پروتئین پوششی ویروس رگبرگ روشنی زرد مرکبات، منجر به تکثیر قطعه‌ مورد نظر با اندازه 614 جفت باز شد. ترادف این ناحیه از ژن در جدایه‌های مورد بررسی با توالی سایر جدایه‌های ویروس رگبرگ روشنی زرد مرکبات از چین، هند، پاکستان و ترکیه در سطح  75/95 تا 37/98 درصد شباهت داشت. درخت تبارزایی بر اساس این قسمت از ژنوم، جدایه‌های ایرانی را با دو جدایه از هند و ترکیه و دو جدایه پاکستانی در یک گروه قرار داد. با توجه به شناسایی ویروس رگبرگ روشنی زرد مرکبات در استان مازندران و نظر به شیوه‌های مختلف انتقال و خسارت بیماری ناشی از این ویروس، پیشگیری از گسترش آن به کانون‌های جدید بسیار حائز اهمیت است.

کلیدواژه‌ها


عنوان مقاله [English]

Identification of Citrus yellow vein clearing virus in Mazandaran province

نویسندگان [English]

  • S. M. Bani Hashemian 1
  • S. Aghajanzadeh 2
1 Assistant Professor, Citrus and Subtropical Fruits Research, Center , Ramsar,,
2 Associate Professor, Citrus and Subtropical Fruits Research Center, Ramsar
چکیده [English]

Background and Objectives
Citrus yellow vein clearing virus (CYVCV), a new member of the genus Mandarivirus (family Alfafelxiviridae), has been recently reported from Iran after being recognized in Pakistan, India, Turkey and China. Typical symptoms of infection on the leaves of sour orange, lemon and Persian lime, as local sensitive hosts, consist of yellowing lateral veins with corresponding water-soaked appearance on the lower surface of the leaves. Biological and molecular detection, some modes of transmission and phylogenetic analysis of CYVCV isolates of Mazandaran province have been investigated in the present study.
Materials and Methods
Typical induced symptoms by CYVCV infection; namely yellowing and water soaking of the lateral veins, was considered as the initial criterion for diagnosis of the disease within seedlings and trees of sour orange, lemon and Persian lime cultivars in nurseries and orchards in Mazandaran province.  Three isolates, out of 53 samples, were selected for more analysis from the region collected during the spring of 2016, 2017, and 2018. Inoculation of these isolates into bean plants, sour oranges and lemons were performed using mechanical and grafting methods under greenhouse conditions with controlled temperatures. In order to confirm the infection, total RNA of the collected samples and the inoculated plants were extracted by the SDS-Potassium acetate method. A two-step Reverse Transcription Polymerase chain reaction (RT-PCR) was carried out using a specific primer pair of the virus coat protein (CP) gene.
Results
Characteristic symptoms of the disease were expressed after top-work grafting of the susceptible virus-free orchard cultivars on the trunks of diseased trees. Using specific primer pair in RT-PCR, an expected 614 bp fragment was amplified in symptomatic samples. The consensus sequences, obtained from direct sequencing the PCR products of the studied isolates based on forward and reverse specific primes, were similar to the sequences of other CYVCV isolates of China, India, Pakistan and Turkey at the level of 95.75 to 98.37%. A phylogenetic tree of this part of the genome, grouped Iranian isolates with two isolates from India and Turkey and two Pakistani isolates.
Discussion
The present study confirmed the presence of Citrus yellow vein clearing virus in the north of the country. Lemon and Persian lime are the two most important susceptible hosts of the virus. These citrus varieties are also valuable cultivars in the south of the country and are considered tolerant genotypes to Witches' broom disease, which is prevalent in these areas. Preventing the spread of the yellow vein clearingdisease to the new infection foci by continuous monitoring of the citrus nurseries should be seriously considered.

کلیدواژه‌ها [English]

  • Citrus yellow vein clearing virus
  • lemon
  • Persian lime
  • Sour orange
Alshami, A.A.A., Ahlawat, Y.S., and Pant, R.P. 2003. A hitherto unreported yellow vein clearing disease of citrus in India and its viral etiology. Indian Phytopathology, 56 (4): 422-427.
Bani Hashemian, S.M. 2012. Current situation of virus and virus-like diseases of citrus trees. 6th Iranian congress of virology, Tehran, Iran. P. 113.
Bani Hashemian, S.M., Taheri, H., Alian, Y.M., Bové, J.M., and Durán-Vila, N. 2013. Complex mixtures of viroids identified in the two main citrus growing areas of Iran. Journal of plant pathology, 95: 647-654.
Bani Hashemian, S.M., and Aghajanzadeh, S. 2017. Occurrence of Citrus yellow vein clearing virus in citrus species in Iran. Journal of Plant Pathology, 99 (1): 290.
Bani Hashemian, S.M. 2018. Citrus yellow vein clearing disease, a new threat for citriculture of Iran. Proceedings of the 23rd Iranian Plant Protection Congress, Gorgan, Iran. P. 626-627.
Bernad, L., and Durán-Vila, N. 2006. A novel RT-PCR approach for detection and characterization of citrus viroids. Molecular and cellular probes, 20 (2): 105-113.
Catara, A., Azzaro, A., Davino, M., and Polizzi, G., 1993. Yellow vein clearing of lemon in Pakistan. Proceedings of the 12th International Organization of Citrus Virologists Conference, New Delhi, India. P. 364-367.
Chen, H.M., Li, Z.A., Wang, X.F., Zhou, Y., Tang, K.Z., Zhou, C.Y., Zhao, X.Y., and Yue, J.Q. 2014. First report of Citrus yellow vein clearing virus on lemon in Yunnan, China. Plant Disease, 98 (12): 1747.
Ebrahimi, Y., and Nematollahi Sani, S. 2003. Top working is a suitable method for improving citrus orchards in northern Iran. Extensional publication of Iran Citrus Research Institute, Ministry of Jihad Agriculture (In Farsi).
Ebrahim-Nesbat, F., and Nienhouse, F. 1978. Occurrence of Citrus tristeza virus in Iran. Zeitschrift für Pflanzenkrankheiten und Pflanzenschutz, 85: 308-312.
FAOSTAT. 2018. Crop statistics of Food and Agriculture Organization. Retrieved December 12, 2019, from http://www.fao.org/faostat.
Grimaldi, V., and Catara, A. 1996. Association of a filamentous virus with yellow vein clearing of lemon. Proceedings of the 13th International Organization of Citrus Virologists Conference, Fouzhou, China. P. 343-345.
Liu, Y., Wang, Y., Wang, Q., Zhang, Y., Shen, W., Li, R., Cao, M., Chen, L., Li, X., Zhou, C., and Zhou, Y. 2019. Development of a sensitive and reliable reverse transcription droplet digital PCR assay for the detection of Citrus yellow vein clearing virus. Archives of virology, 164 (3): 691-697.
Loconsole, G., Önelge, N., Potere, O., Giampetruzzi, A., Bozan, O., Satar, S., De Stradis, A., Savino, V., Yokomi, R.K., and Saponari, M. 2012. Identification and characterization of Citrus yellow vein clearing virus, a putative new member of the genus Mandarivirus. Phytopathology, 102 (12): 1168-1175.
Önelge, N. 2002. First report of yellow vein clearing of lemons in Turkey. Journal of Turkish Phytopathology, 32: 53-55.
Önelge, N., Bozan, O., Gök, M., and Satar, S. 2010. Yellow vein clearing of lemons in Turkey. Proceedings of the 17th International Organization of Citrus Virologists Conference, Adana, Turkey. P. 227-228.
Önelge, N., Satar, S., Elibüyük, Ö., and Bozan, O. 2011a. Citrus yellow vein clearing virus: A new aphid-transmitted citrus virus. Citrograph, 1: 22-24.
Önelge, N., Satar, S., Elibüyük, Ö., Bozan, O., and Kamberoolu, M. 2011b. Transmission studies on Citrus yellow vein clearing virus. Proceedings of the 18th International Organization of Citrus Virologists Conference, Sao Paulo, Brazil. P. 11-14.
Nasrollanejad, S., Ziyarat, M.A., and Fakhrabad, F.Z. 2012. The determination of Citrus tristeza virus strains in the North of Iran. International Journal of Agriculture and Crop Sciences, 4 (22): 1714-1719.
Roistacher, C.N. 1991. Graft-transmissible diseases of citrus: Handbook for detection and diagnosis. FAO publications, Rome.
Salehi, M., Nejat, N., Tavakkoli, A.R., and Izadpanah, K., 2005. Reaction of citrus cultivars to Candidatus phytoplasma aurantifolla in Iran. Iranian Journal of Plant Pathology, 41: 363-376 (In Farsi with English summary).
Tarviji, M., Bani Hashemian, S.M., Elahinia, S.A., and Ebrahimi Moghaddam, L. 2012. Biological and molecular characterization of two Citrus tristeza virus isolates obtained from west part of Mazandaran province. Proceeding of the 20th Iranian Plant Protection Congress. Shiraz, Iran. P. 888.
Zafari, H., Bani Hashemian, S.M., Ruhibakhsh, A., and Bahri, B. 2013. First report of Citrus tristeza virus from Guilan province. Iranian Journal of Plant Pathology, 49: 175-176 (In Farsi with English summary).
Zhang, Y., Wang, Y., Wang, Q., Cao, M., Zhou, C., and Zhou, Y. 2018. Identification of Aphis spiraecola as a vector of Citrus yellow vein clearing virus. European Journal of Plant Pathology, 152 (3): 841-844.
Zhang, Y.H., Liu, C.H., Wang, Q., Wang, Y.L., Zhou, C.Y., and Zhou, Y. 2019a. Identification of Dialeurodes citri as a vector of Citrus yellow vein clearing virus in China. Plant disease, 103 (1): 65-68.
Zhang, Y., Liu, Y., Wang, Y., Wang, Q., He, S., Li, X., and Zhou, Y. 2019b. Transmissibility of Citrus yellow vein clearing virus by contaminated tools. Journal of Plant Pathology, 101 (1): 169-171.
Zhou, Y., Chen, H.M., Wang, X.F., Li, Z.A., Tang, M., and Zhou, C.Y. 2015. Lack of evidence for seed transmission of Citrus yellow vein clearing virus despite its frequent detection in seed tissues. Journal of Plant Pathology, 97: 1-3.
Zhou, Y., Chen, H.M., Cao, M.J., Wang, X.F., Jin, X., Liu, K.H., and Zhou, C.Y. 2017. Occurrence, distribution, and molecular characterization of Citrus yellow vein clearing virus in China. Plant disease, 101 (1): 137-143.