اهمیت و ویژگی های مولکولی ویروس موزائیک خیار مزارع جالیزی جهرم
محورهای موضوعی : ویروس شناسی
کاووس ایازپور
1
(هیات علمی، دانشگاه آزاد اسلامی، واحد جهرم)
سیده زهرا ساجدی
2
(گروه بیماری شناسی گیاهی، واحد جهرم، دانشگاه آزاد اسلامی، جهرم، ایران)
کلید واژه: DAS-ELISA, آنالیز فیلوژنتیکی, کوکوربیتاسه,
چکیده مقاله :
سابقه و هدف: ویروس موزاییک خیار( (CMV یک کوکوموویروس با انتشار جهانی و یکی از ویروس های مهم کدوئیان و سایر محصولات است که سبب کاهش قابل توجه محصول می گردد. این پژوهش به منظور به دست آوردن اطلاعاتی در مورد انتشار، اهمیت و ویژگی مولکولی CMV در مزارع جالیزی منطقه جهرم انجام شد.مواد و روش ها: 147 نمونه گیاهی مشکوک به علایم CMV از برگ های بوته های ارقام مختلف خیار، کدو، هندوانه و خربزه از مزارع مختلف کدوئیان و گلخانه های خیار شهرستان جهرم جمع آوری و با استفاده از آزمون سرولوژیکی الایزا(DAS-ELISA) و آزمون مولکولی RT-PCR مورد بررسی قرار گرفتند. محصولات PCR مربوط به دو جدایه از روستاهای قطب آباد و هورموج تعیین توالی و با 18 توالی NCBI مقایسه شدند. سپس درخت فیلوژنتیکی با کمک نرم افزار های ClustalW و MEGA6 با روش Neighbor-Joining ترسیم گردید.یافته ها: با استفاده از آزمون الیزا 59 نمونه آلوده به CMV تشخیص داده شد. واکنش RT-PCR نیز آلودگی ها را تأیید نمود. آنالیز توالی ها نشان داد که دو جدایه CMV-JS1و CMV-JS2شهرستان جهرم همراه با جدایه های M21464.1، AJ242585، AJ866272.1،L40953.1 به ترتیب از استرالیا، چین، هند و آفریقای جنوبی در یک زیر گروه قرار می گیرند.نتیجه گیری: نتایج نشان داد که آلودگی قابل توجهی به ویروس موزائیک خیار در مزارع جالیزی منطقه جهرم وجود دارد که می تواند کاهش محسوس محصول را به دنبال داشته باشد. به دلیل دامنه میزبانی وسیع CMV و امکان انتقال به وسیله شته ها، از این رو توصیه می گردد که علاوه بر کنترل علف های هرز در مزارع مبارزه با شته ها نیز انجام شود.
Background and Objectives: The Cucumber Mosaic Virus (CMV) is a worldwide distributed Cucumovirus, and one of the important viruses of cucurbits and other agricultural crops, which causes a significant reduction of yield. This study was done to determine some information about the distribution, importance, and molecular characteristics of CMV in cucurbitaceous fields of the Jahrom area. Material and Methods: 147 plant samples suspected of CMV symptoms were collected from leaves of different varieties of cucumber, pumpkin, watermelon, and melons from various farms of cucurbits and cucumber greenhouses of the Jahrom area and were investigated using DAS-ELISA and RT-PCR methods. The PCR products of two-isolates were sequenced from the Ghotbabad and Hormoj villages. These isolates sequences were compared with 19 selected sequences of NCBI. Using Clustal W and Mega6 software, the phylogenetic tree was drawn via the Neighbor-Joining method. Results: 59 samples were detected as CMV infected by DAS-ELISA test. The RT-PCR reaction confirmed the infection. Sequences analysis showed that the two isolates of CMV-JS1 and CMV-JS2 from the Jahrom area with the isolates of M21464.1, AJ242585, AJ866272.1, and L40953.1, respectively from Australia, China, India, and South Africa are located in an individual subgroup. Conclusion:The results of this study showed that there where a significant infection in cucurbit fields of the Jahrom area and may cause significant yield losses. Since the CMV has a wide host range and is transmitted by aphids, hence it is recommended to control weeds as well as vector aphids.
1992; 414: 281-348.
2. Roossinck MJ. Cucumber Mosaic Virus, a model for RNA virus evolution. Mol Plant Pathol.
2001; 2: 59–63.
3. Yoshida K, Goto T, Lizuka N. Attenuated isolates of Cucumber Mosaic Virus produced by
satellite RNA and cross-protection between attenuated and virulent ones. Ann Phytopathol Soc
Japan. 1985; 51: 238-242.
4. Gallitelli D, Di Franco A, Vovlas C, Kaper JM. Infezioni miste del virus del mosaico del
cetriolo (CMV) e di potyvirus in colture ortive di Puglia e Basilicata. Inf Fitopatol. 1988; 12:
57–64.
5. Kaper JM, Gallitelli D, Tousignant ME. Identification of a 334-ribonucleotid viral satellite as
principal a etiological agent in a tomato necrosis epidemic. Res Virol. 1990; 141:81-95.
1. Jorda C, Alfaro A, Aranda MA, Moriones E, Garcıa-Arenal F. Epidemic of Cucumber Mosaic
Virus plus satellite RNA in tomatoes in Eastern Spain. Plant Dis. 1992; 76: 363–366.
1. Jones RAC, Proudlove W. Further studies on cucumber mosaic virus infection of narrow-leafed
lupin (Lupinus angustifolius): seed-borne infection, aphid transmission, spread and effects on
grain yield. Ann Appl Biol. 1991; 118, 319–329.
8. Tepfer M, Girardot G, Feneant L, Tamarzizt HB, Verdin E, Moury BT, Jacquemond M.
A genetically novel, narrow-host-range isolate of Cucumber Mosaic Virus (CMV) from
rosemary. Arch Virol. 2016; 161: 2013-2017.
1. Ohshima K, Matsumoto K, Yasaka R, Nishiyama M, Soejima K, Korkmaz S, Ho SY, Gibbs AJ,
Takeshita M. Temporal analysis of reassortment and molecular evolution of Cucumber mosaic
virus: Extra clues from its segmented genome. 2016; Virol. 487: 188-197.
10. Hasiow-Jaroszewska B, Chrzanowski M, Budzynska D, Rymelska N, Borodynko-Filas N.
Genetic diversity, distant phylogenetic relationships and the occurrence of recombination
events among cucumber mosaic virus isolates from zucchini in Poland. Arch Virol. 2017; 162:
1751-1756.
11. Choi SK, Choi JK, Park WM, Ryu KH. RT-PCR detection and identification of three species
of cucumoviruses with a genus-specific single pair of primers. J Virol Methods. 1999; 83:
67-73.
12. Shi L, Yang Y, Xie Q, Miao H, Bo K, Song Z, Wang Y, Xie B, Zhang S, Gu X. Inheritance
and QTL mapping of Cucumber Mosaic Virus resistance in cucumber (Cucumis Sativus L.).
PLoS One. 2018; 13: e0200571. https://doi.org/10.1371/journal.pone.0200571.
13. Palukaitis P, Garcia-Arenal F. Cucumoviruses. Advances in Virus Research. 2003; 62:
241-323.
14. Tomlinson JA, Carter AL. Studies on the seed transmission of Cucumber Mosaic Virus in
chickweed, Stellaria media, in relation to the ecology of the virus. Ann Appl Biol. 1970;
66:381-386.
15. Seo YS, Rojas MR, Lee JY, Lee SW, Jeon JS, Ronald P, Lucas WJ, Gilbertson RL. A viral
resistance gene from common bean functions across plant families and is up-regulated in a
non-virus-specific manner. Proc Natl Acad Sci USA. 2006; 103 (32): 11856–11861.
11. Wang HL, Sudarshana MR, Gilbertson RL, Lucas WJ. Analysis of cell-to-cell and
long-distance movement of a bean dwarf mosaic geminivirus-green fluorescent protein reporter
in host and non-host species: identification of sites of resistance. Mol Plant Microbe Interact.
1999; 12: 345–355.
11. Ayazpour K. Alphabetic lists of plant viruses and viroids reported from Iran. Jahrom. Jahrom
Branch, Islamic Azad University; 2014.
18. Clark MF, Adams AN. Characteristics of microplate method of enzyme-linked
immunosorbent assay for detection of plant viruses. J Gen Virol. 1977; 34: 475-483.
11. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics
Analysis across computing platforms. Mol Biol Evol. 2018; 35:1547-1549.
20. Zollanvari Z, Ghasemi S, Pakniat A. Detection of cucurbits infecting viruses in Shiraz zone
and suburbs by using serological and molecular methods. Plant Protec J. 2012; 4: 95-106 (In
Persian).
21. Samei A, Massumi H, Shaabanian M, Hosseini Pour A, Heydarnejad J. Evaluation of some
cucurbit cultivars grown in the fields and greenhouses to six important viruses. J Agric Sci Nat
Resou. 2008; 15:1-8 (In Persian).
22. Sydanmetsa M, Mbanzibwa DR. Occurrence of Cucumber Mosaic Virus, Zucchini Yellow
Mosaic Virus and Watermelon Mosaic Virus in cultivated and wild cucurbits in the coastal
areas of Tanzania. Afr J Agric Res. 2016; 11: 4062-4069.
23. Sokhandan NB, Nematollahi S, Torabi E. Cucumber Mosaic Virus subgroup IA frequently
occurs in the northwest Iran. Acta Virol. 2008; 52: 237-242.
24. Ayazpour K, Vahidian M. Study of Watermelon Mosaic Virus in Cucurbit Fields of Jahrom
Area, Iran. Vegetos. 2016; 29: DOI: 10.4172/2229-4473.1000171.
25. Demeski W, Sowell G. Susceptibility of Cucurbita pepo and Citrullus lanatus introduction to
Watermelon Mosaic Virus-2. Plant Dis. 1970; 54: 880-881.
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1992; 414: 281-348.
2. Roossinck MJ. Cucumber Mosaic Virus, a model for RNA virus evolution. Mol Plant Pathol.
2001; 2: 59–63.
3. Yoshida K, Goto T, Lizuka N. Attenuated isolates of Cucumber Mosaic Virus produced by
satellite RNA and cross-protection between attenuated and virulent ones. Ann Phytopathol Soc
Japan. 1985; 51: 238-242.
4. Gallitelli D, Di Franco A, Vovlas C, Kaper JM. Infezioni miste del virus del mosaico del
cetriolo (CMV) e di potyvirus in colture ortive di Puglia e Basilicata. Inf Fitopatol. 1988; 12:
57–64.
5. Kaper JM, Gallitelli D, Tousignant ME. Identification of a 334-ribonucleotid viral satellite as
principal a etiological agent in a tomato necrosis epidemic. Res Virol. 1990; 141:81-95.
1. Jorda C, Alfaro A, Aranda MA, Moriones E, Garcıa-Arenal F. Epidemic of Cucumber Mosaic
Virus plus satellite RNA in tomatoes in Eastern Spain. Plant Dis. 1992; 76: 363–366.
1. Jones RAC, Proudlove W. Further studies on cucumber mosaic virus infection of narrow-leafed
lupin (Lupinus angustifolius): seed-borne infection, aphid transmission, spread and effects on
grain yield. Ann Appl Biol. 1991; 118, 319–329.
8. Tepfer M, Girardot G, Feneant L, Tamarzizt HB, Verdin E, Moury BT, Jacquemond M.
A genetically novel, narrow-host-range isolate of Cucumber Mosaic Virus (CMV) from
rosemary. Arch Virol. 2016; 161: 2013-2017.
1. Ohshima K, Matsumoto K, Yasaka R, Nishiyama M, Soejima K, Korkmaz S, Ho SY, Gibbs AJ,
Takeshita M. Temporal analysis of reassortment and molecular evolution of Cucumber mosaic
virus: Extra clues from its segmented genome. 2016; Virol. 487: 188-197.
10. Hasiow-Jaroszewska B, Chrzanowski M, Budzynska D, Rymelska N, Borodynko-Filas N.
Genetic diversity, distant phylogenetic relationships and the occurrence of recombination
events among cucumber mosaic virus isolates from zucchini in Poland. Arch Virol. 2017; 162:
1751-1756.
11. Choi SK, Choi JK, Park WM, Ryu KH. RT-PCR detection and identification of three species
of cucumoviruses with a genus-specific single pair of primers. J Virol Methods. 1999; 83:
67-73.
12. Shi L, Yang Y, Xie Q, Miao H, Bo K, Song Z, Wang Y, Xie B, Zhang S, Gu X. Inheritance
and QTL mapping of Cucumber Mosaic Virus resistance in cucumber (Cucumis Sativus L.).
PLoS One. 2018; 13: e0200571. https://doi.org/10.1371/journal.pone.0200571.
13. Palukaitis P, Garcia-Arenal F. Cucumoviruses. Advances in Virus Research. 2003; 62:
241-323.
14. Tomlinson JA, Carter AL. Studies on the seed transmission of Cucumber Mosaic Virus in
chickweed, Stellaria media, in relation to the ecology of the virus. Ann Appl Biol. 1970;
66:381-386.
15. Seo YS, Rojas MR, Lee JY, Lee SW, Jeon JS, Ronald P, Lucas WJ, Gilbertson RL. A viral
resistance gene from common bean functions across plant families and is up-regulated in a
non-virus-specific manner. Proc Natl Acad Sci USA. 2006; 103 (32): 11856–11861.
11. Wang HL, Sudarshana MR, Gilbertson RL, Lucas WJ. Analysis of cell-to-cell and
long-distance movement of a bean dwarf mosaic geminivirus-green fluorescent protein reporter
in host and non-host species: identification of sites of resistance. Mol Plant Microbe Interact.
1999; 12: 345–355.
11. Ayazpour K. Alphabetic lists of plant viruses and viroids reported from Iran. Jahrom. Jahrom
Branch, Islamic Azad University; 2014.
18. Clark MF, Adams AN. Characteristics of microplate method of enzyme-linked
immunosorbent assay for detection of plant viruses. J Gen Virol. 1977; 34: 475-483.
11. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics
Analysis across computing platforms. Mol Biol Evol. 2018; 35:1547-1549.
20. Zollanvari Z, Ghasemi S, Pakniat A. Detection of cucurbits infecting viruses in Shiraz zone
and suburbs by using serological and molecular methods. Plant Protec J. 2012; 4: 95-106 (In
Persian).
21. Samei A, Massumi H, Shaabanian M, Hosseini Pour A, Heydarnejad J. Evaluation of some
cucurbit cultivars grown in the fields and greenhouses to six important viruses. J Agric Sci Nat
Resou. 2008; 15:1-8 (In Persian).
22. Sydanmetsa M, Mbanzibwa DR. Occurrence of Cucumber Mosaic Virus, Zucchini Yellow
Mosaic Virus and Watermelon Mosaic Virus in cultivated and wild cucurbits in the coastal
areas of Tanzania. Afr J Agric Res. 2016; 11: 4062-4069.
23. Sokhandan NB, Nematollahi S, Torabi E. Cucumber Mosaic Virus subgroup IA frequently
occurs in the northwest Iran. Acta Virol. 2008; 52: 237-242.
24. Ayazpour K, Vahidian M. Study of Watermelon Mosaic Virus in Cucurbit Fields of Jahrom
Area, Iran. Vegetos. 2016; 29: DOI: 10.4172/2229-4473.1000171.
25. Demeski W, Sowell G. Susceptibility of Cucurbita pepo and Citrullus lanatus introduction to
Watermelon Mosaic Virus-2. Plant Dis. 1970; 54: 880-881.