Detection and Characterization of Tomato spotted wilt virus and Cucumber mosaic virus on Pepper Growing Areas in Antalya

Abstract

The most efficient method to control the plant virus diseases is breeding resistant cultivars. However, the resistance could be broken down after using resistant cultivars. This study was aimed to determine the prevalence and also serological and molecular characterization of Tomato spotted wilt virus (TSWV) and Cucumber mosaic virus (CMV) that cause infections, especially, in resistant pepper cultivars. For this reason, samples were collected from pepper growing greenhouses and open fields during vegetation period of 2015 in different parts of Antalya province including Kumluca, Demre, Serik and Aksu districts. Out of 148 pepper samples collected, 53 (35.81%) were infected with TSWV and 11 (7.34%) with CMV as a result of Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DASELISA) test. Some regions on S RNA (nucleocapsid protein gene), M RNA (glycoprotein gene) and L RNA (RNAdependent RNA polymerase gene) of TSWV genome; RNA 1 (helicase/methyltransferase gene) and RNA 3 (coat protein

gene) of CMV genome of DAS-ELISA positive samples were amplified by RT-PCR with specific primers. Nucleotide similarity rates of nucleocapsid protein gene, glycoprotein gene and RNA-dependent RNA polymerase gene regions of TSWV isolate varied between 92-98% identity with other isolates in GenBank and CMV isolate varied between 89-96%. TSWV isolate showed nucleotide identity varied between 92-97% with Tsw resistance breaking isolates from Capsicum plants in Turkey, Italy and Spain. Genetic determinant for overcoming pepper Tsw resistance is located in S segment but aminoacid substitutions responsible for TSWV breakdown remain contradictory in several reports.

Keywords

• Cucumber mosaic virus (CMV)
• Tomato spotted wilt virus (TSWV)
• DAS-ELISA
• RT-PCR
• Detection
• Characterization

References

1. Adkins S (2000). Tomato spotted wilt virus-positive steps towards negative success, Molecular Plant Pathology, 1(3):151-157 pp.
2. Arli-Sokmen M A, Mennan H, Sevik M A & Ecevit O (2005). Occurrence of viruses in field-grown pepper crops and some of their reservoir weed hosts in Samsun, Turkey, Phytoparasitica, 33(4):347-358pp.
3. Biswas K, Hallan V, Zaidi A A & Pandey P K (2013). Molecular evidence of Cucumber mosaic virus subgroup II infecting Capsicum annuum L. in the Western region of India, Current Discovery, 2(2):97-105 pp.
4. Burgmans J L, Fry P R & Sunde R G (1986). Peppers: survey of virus diseases of Capsicum annuum in Hawke’s Bay and Poverty Bay, New Zealand Journal of Experimental Agriculture, 14:459-463 pp.
5. Buzkan N, Demir M, Öztekin V, Mart C, Çaǧlar B K & Yilmaz M A (2006). Evaluation of the status of capsicum viruses in the main growing regions of Turkey, EPPO Bulletin, 36:15-19 pp.
6. De Blas C, Borza M J, Saiz M & Romero J (1994). Broad spectrum detection of Cucumber mosaic virus (CMV) using the polymerase chain reaction, Journal of Phytopathology, 141:323-329 pp.
7. Deligoz I, Sokmen M A & Sari S (2014). First report of resistance breaking strain of Tomato spotted wilt virus (Tospovirus; Bunyaviridae) on resistant sweet pepper cultivars in Turkey, New Disease Reports, 30:26 p.
8. Ferrand L, García M L, Resende R O, Balatti P A & Dal Bó E (2015). First report of a resistance-breaking isolate of Tomato spotted wilt virus infecting sweet pepper harboring the Tsw gene in Argentina, Plant Disease, 99:12 p.

9. Foissac X, Svanella-Dumas L, Dulucq M J, Candresse T & Gentit P (2001). Polyvalent detection of fruit tree tricho, capillo and foveaviruses by nested RT-PCR using degenerated and inosine containing primers (PDO RT-PCR), Acta Hortic, 550:37-43 pp.
10. Gabor B, Krizbai L, Horvath J & Takacs A (2012). Resistance breaking strain of Tomato spotted wilt virus (TSWV) on resistant pepper cultivars in Hungary, Proceedings of the International Symposium on Current Trends in Plant Protection, Belgrade, Serbia, 239-241 pp.
11. Kumari R, Bhardwaj P, Singh L, Zaidi A A & Hallan V (2013). Biological and molecular characterization of Cucumber mosaic virus subgroup II isolate causing severe mosaic in cucumber, Indian Journal of Virology, 24(1):27-34 pp.
12. Margaria P, Ciuffo M & Turina M (2004). Resistance breaking strain of TSWV Tospovirus, (Bunyaviridae) on resistant pepper cultivars in Almeria Spain, Plant Pathology, 53:795 p.
13. Mavric I & Ravnikar M (2001). First report of Tomato spotted wilt virus and Impatient necrotic spot virus in Slovenia, Plant Disease, 85(12):1288 p.
14. Mound L A (2001). So many thrips–so few tospoviruses, Thrips and Tospoviruses: Proceedings of the VII. International Symposium on Thysanoptera, Reggio Calabria, 15-18 pp.
15. Moury B & Verdin E (2012). Viruses of pepper crops in the Mediterranean basin: a remarkable stasis, Advances in Virus Research, 84:127-162 pp.
16. Nour S M, Maleki M & Ghotbi T (2013). Biological and serological detection of TSWV on three commercial cultivars Chrysanthemum morifolium in Markazi province of Iran, Annals of Biological Research, 4(4):112-119 pp.
17. Perry K L, Zhang L & Palukaitis P (1998). Amino acid changes in the coat protein of Cucumber mosaic virus differentially affect transmission by the aphids Myzus persicae and Aphis gossypii, Virology, 242:204-210 pp.
18. Roggero P, Masenga V & Tavella L (2002). Field isolates of Tomato spotted wilt virus overcoming resistance in pepper and their spread to other hosts in Italy, Plant Disease, 86:950-954 pp.
19. Sharman M & Persley D M (2006). Field isolates of Tomato spotted wilt virus overcoming resistance in Capsicum in Australia, Australasian Plant Pathology, 35:123-128 pp.
20. Uzunoğulları N & Gümüş M (2015). Marmara bölgesi’nde bazı kültür bitkilerinde doğal enfeksiyona neden olan hıyar mozaik virüsü (Cucumber mosaic virus, CMV)’nün tespiti, Trakya University Journal of Natural Sciences, 16(1):9-15 pp.
21. Vozelj N, Petrovic N, Novak M P, Tusek M, Mavric I & Ravnikar M (2003). The most frequent viruses on selected ornamental plants and vegetables in Slovenia, Zbornik predavanj in referatov 6.Slovenskega Posvetovanje o Varstvu Rastlin, Zrece, 300-304 pp.
22. Yardımcı N & Çulal Kılıç H (2009). Tomato spotted wilt virus in vegetable growing areas in the West Mediterranean region of Turkey, African Journal of Biotechnology, 8(18):4539-4541 p.
23. Yılmaz M A & Davis R F (1985). Identification of viruses infecting vegetable crops along the Mediterranean Sea coast in Turkey, The Journal of Turkish Phytopathology,14:1-8 pp.