Molecular detection of Cucumber vein yellowing virus and Cucurbit yellow stunting disorder virus in southeastern Iran and the genetic analysis of isolates from CVYV

Document Type : Research Paper

Authors

1 Ph. D. Student of Virology and Plant Viral Diseases, Department of Plant Protection, Gorgon University of Agricultural Sciences & Natural Resources, Gorgon, Iran.

2 Associate Professor, Department of Plant Protection, Gorgon University of Agricultural Sciences & Natural Resources, Gorgon, Iran.

3 Assistant Professor, Department of Biology, University of Sistan and Baluchestan, Zahedan, Iran.

4 Assistant Professor, Department of Plant Protection, Gorgon University of Agricultural Sciences & Natural Resources, Gorgon, Iran.

5 Assistant Professor, Department of Plant Protection, Higher Educational Complex of Saravan, Saravan, Iran.

Abstract

Yellowing diseases of field and greenhouse-grown cucurbits caused by whitefly-transmitted viruses are increasingly becoming important and cause economic losses in many cucurbits growing areas of the world. In this research, 195 samples with symptoms yellowing, stunting and deformation of leaves were collected from fields of cucurbits in the South-East Iran (Sistan and Baluchestan). In order to detect CVYV and CYSDV, cDNAs were prepared using Random Hexamer primer and total RNAs extracted from the collected samples.Then, Polymerase Chain Reaction (PCR) was performed using exteracted RNA and specific primers (CYSCPf and CYSCPr for CYSDV, CV+ and CV- for CVYV). The results revealed that a 450 bp fragment from the CVYV-CP and a 700 bp fragment from CYSDV-CP were amplified from 55 and 17 samples, respectively. About 7% of the samples had simultaneous contamination with the two viruses. Based on the Phylogenetic studies CVYV were divided into two main groups I (subgroups IA and IB) and II: the first group belongs of isolates from Lebanon and Jordan, Tunisia and Spain, but the second group contained only isolates of Iran (Chabahar) which indicates the high diversity of this isolate. The results of the genetic diversity and selection indices showed that the gene flow from Europe to Asia is very small and the genetic difference is high between them. The CP region is under severe pressure a negative choice, and has caused variation and evolution of the CVYV. Generally, based on the results, the most diversity molecular and host are in Asia group.

Keywords

Main Subjects


Abrahamian PE, Abou-Jawdah, Y (2014) Whitefly-transmitted criniviruses of cucurbits: current status and future prospects. Virus Disease. 25: 26-38.
Ahmadi K, Ebadzadeh HR, Abdi Shah H, Kazemian A, Rafiei M (2017) Agricultural statistics crop year 2016-2017. Ministry of Agriculture Jahad, Iran.
Bananej K, Kianfar N, Winter S, Menzel W (2014) The status of Cucumber vein yellowing virus in Iran. Phytopathologia Mediterranea. 53: 269-276.
Cohen S, Nitzany FE (1960) A whitefly-transmitted virus of cucurbits. Phytopathology Mediterranea. 1: 44-46.
Cuadrado IM, Janssen D, Velasco L, Ruiz L, Segundo E (2001) First report of Cucumber vein Yellowing virus in Spain. Plant Disease. 85: 336.
Desbiez C, Lecoq H, Girard M, Cotillon AC, Schoen L (2003) First report of Cucurbit yellow stunting disorder virus in commercial cucumber greenhouses in France. Plant Disease. 87: 600.     
De Bokx, Van der JA, Want JPH (1987) Viruses of potato and seed-potato production, 2nd Ed. Center for Agricultural Publishing and Documentation, Pudoc., Wageningen, Netherlands, 259 pp.
Gil-Salas FM, Peters J, Boonham N, Cuadrado IM, Janssen D (2011) Yellowing disease in zucchini squash produced by mixed infections of Cucurbit yellow stunting disorder virus and Cucumber vein yellowing virus. Phytopathology 101:1365-1372.
Hull R (2014) Comparative plant virology. 2th ed. Academic press.
Janssen D, Ruiz L, Velasco L, Segundo E, Cuadrado IM (2002) Non-cucurbitaceous weed species shown to be natural hosts of Cucumber vein yellowing virus in south-eastern Spain. Plant Pathology. 51: 797.
Janssen D, Ruiz L, Cano M, Belmonte A, Martin G, Segundo E, Cuadrado IM (2003) Physical and genetic control of Bemisia tabaci-transmitted Cucurbit yellow stunting disorder virus and Cucumber vein yellowing virus in cucumber. International Organisation for Biological and integrated Control (west palaearctic regional section) Bulletin 26: 101-6.
Janssen D, Martín G, Velasco L, Cuadrado IM, Segundo E (2007) Low genetic diversity among Cucumber vein yellowing virus isolates from Spain. Virus Genes. 34: 367–371.
Kosakovsky Pond SL, Frost SDW, Muse SV (2005) HyPhy: hypothesis testing using phylogenies. Bioinformatics. 21: 676-679.
Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution. 33: 1870-1874.
Lecoq H, Desbiez C (2012) Advances in virus research In: Gad L, Herve´ L (ed) Viruses of cucurbit crops in the Mediterranean region: an ever-changing picture, Academic Press, London, pp 67-126.
Mansour A, Al-Musa A (1993) Cucumber vein yellowing virus; host range and virus vector relationships. Journal of Phytopathology. 137: 73-78.
Pic´o B, Sifres A, Nuez, F (2005) Quantitative detection of Cucumber vein yellowing virus in susceptible and partially resistant plants using real-time PCR. Journal of Virological Methods. 128: 14-20.
Rozas J, Ferrer-Mata A, Sanchez-DelBarrio JC, Guirao-Rico S, Librado P, Ramos-Onsins SE, Sanchez-Gracia A (2017) DnaSP v6: DNA Sequence Polymorphism Analysis of Large Datasets. Molecular Biology and Evolution. 34: 3299-3302.
Rubio L, Abou-Jawdah Y, Lin H-X, Falk BW (2001) Geographically distant isolates of the crinivirus Cucurbit yellow stunting disorder virus show very low genetic diversity in the coat protein gene. Journal General Virology. 82: 929-33.
Ruiz L, Janssen D, Martín G, Velasco L, Segundo E, Cuadrado IM (2006) Analysis of the temporal and spatial disease progress of Bemisia tabaci-transmitted Cucurbit yellow stunting disorder virus And Cucumber vein yellowing virus in cucumber. Plant Pathology. 55: 264-275.
Sanchez-Campos S, Dı´az JA, Monci F, Bejarano ER, Reina J, Navas-Castillo J, Aranda MA Moriones E (2002) High genetic stability of the begomovirus Tomato yellow leaf curl Sardinia virus in southern Spain over an 8-year period. Phytopathology. 92: 842-849.
Samei A, Massumi H, Shaabanian M, Hosseinipour A, Heydarnejad J (2008) Evaluation of some cucurbit cultivars grown in the fields and greenhouses to six important viruses. Journal of Agricultural Sciences and Natural Resources. 15: 4-12.
Shoeibi S, Nasrollanezhad S (2009) Detection and Distribution of Watermelon mosaic virus (WMV) in cucurbits fields of Golestan province Journal of Agricultural Sciences and Natural Resources. 16: 1-4.
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties, and weight matrix choice. Nucleic Acids Research. 22: 4073-4080.
Yakoubi S, Desbiez C, Fakhfakh H, Wipf-Scheibel C, Marrakchi M Lecoq H (2007) Occurrence of Cucurbit yellow stunting disorder virus and Cucumber vein yellowing virus in Tunisia. Journal of Plant Pathology. 89: 417-20.