Comparative analysis of mechanical inoculation methods for Tomato spotted wilt orthotospovirus (TSWV) in tomato

Year 2025, Volume: 38 Issue: 1, 1 - 7, 09.04.2025

Yunus Emre Uslu , Melda Ertürk , Sevdiye Yorgancı

https://doi.org/10.29136/mediterranean.1518443

Abstract

Tomato spotted wilt orthotospovirus (TSWV) is a pathogen that causes significant yield and quality losses in 1000 plant species in 84 families, including tomato, pepper, lettuce, tobacco, and various weeds. TSWV, a member of the Orthotospovirus genus within the Bunyaviridae family, is characterized by an isometrically enveloped particle. The virus is transmitted both mechanically and via vectors. This study evaluated the effects of two different mechanical inoculation methods, each using a different buffer solution, for in vitro TSWV inoculation. Tomato plants germinated from seeds were infected with TSWV using these two distinct mechanical inoculation methods, and infection was subsequently confirmed via PCR. The presence of TSWV in the infected plants was further validated by Sanger sequencing. Disease incidence and quantitative characteristics of TSWV-infected tomato plants were measured and statistically analyzed to compare the efficiency of each inoculation method and buffer combination. Differences in disease incidence rates and quantitative properties were observed between the two inoculation methods. One of the mechanical inoculation methods, in combination with its respective buffer, demonstrated a 30% higher transmission efficiency for TSWV, emphasizing the impact of both inoculation technique and buffer composition on successful virus transmission.

Keywords

Tomato spotted wilt Orthotospovirus, Tomato, Mechanic inoculation, Physical parameters

References

• Aksu G, Temel E, and Altay H (2017) Yaprak Gübrelemesi ile Roka (Eruca vesicaria) Bitkisinin İyotça Zenginleştirilmesi. Canakkale 18 Mart University Journal of Agriculture Faculty 5: 97-104.
• Alam A, Elçi E (2021) Evaluation of Eggplant Cultivars for Tomato Spotted Wilt Orthotospovirus (TSWV) Disease Tolerance in Greenhouse Conditions. International Journal of Agriculture, Environment, and Food Sciences 5: 497-507.
• Bora T, Karaca İ (1970) Kültür bitkilerinde hastalığın ve zararın ölçülmesi. Ege Üniversitesi Yardımcı Ders Kitabı, 167: 239-248.
• Caruso A, Ragona A, Agrò G, Bertacca S, Yahyaoui E, Galipienso L, Rubio L, Panno S, Davino S (2024) Rapid detection of tomato spotted wilt virus by real-time RT-LAMP and in-field application. Journal of Plant Pathology 106: 697-712.
• Coutts B, Jones R (2005) Suppressing spread of Tomato spotted wilt virus by drenching infected source or healthy recipient plants with neonicotinoid insecticides to control thrips vectors. Annals of Applied Biology 146: 95-103.
• de Haan P, Wagemakers L, Peters D, Goldbach R (1990) The S RNA segment of tomato spotted wilt virus has an ambisense character. The Journal of General Virology 71: 1001-1007.
• Dianese E, Fonseca M, Inoue-Nagata A Resende, and Boiteux L (2011) Search in Solanum (section Lycopersicon) germplasm for sources of broad-spectrum resistance to four Tospovirus species. Euphytica 180: 307-319.
• Erilmez S, Oz O (2023) First report of tomato spotted wilt virus in Zinnia elegans in Turkey. Journal of Plant Pathology 105: 609-609.
• Farooq A, Akanda A (2007) Symptoms and prevalence of Tomato Spotted Wilt Virus (TSWV) infection in Bangladesh. International Journal of Sustainable Crop Production 2: 51-58.
• Fidan H, Sari N (2019) Molecular characterization of resistance-breaking Tomato spotted wilt virus (TSWV) isolate medium segment in tomato. Applied Ecology and Environmental Research 17: 5321-5339.
• Food and Agriculture Organization (FAO) (2022) Tomato production in the world. FAO. Retrieved 2022.
• Gordillo LF, Stevens MR, Millard MA, and Geary B (2008) Screening Two Lycopersicon peruvianum Collections for Resistance to Tomato spotted wilt virus. Plant Disease 92: 694-704.
• Güldür ME, Marcoux G, Yürtmen M, Yılmaz MA (1995) Mersin ve çevresinde yetiştirilen domateslerde zararlı yeni bir virüs: Tomato spotted wilt virus. VII. Fitopatoloji Kongresi Adana, Türkiye 26(29): 303-305.
• Hossain M, Akanda M, Hossain M, and Ahmed J (2016) Yield Components in TSWV Infected Tomato (Solanum lycopersicum L.). International Journal of Business, Social and Scientific Research 4: 127-131.
• Hull R (2009) Mechanical inoculation of plant viruses. Current Protocols in Microbiology 13(1): 16-11.
• Lian S, Lee JS, Cho WK, Yu J, Kim MK, Choi HS, Kim KH (2013) Phylogenetic and Recombination Analysis of Tomato Spotted Wilt Virus. PLOS ONE 8(5): e63380.
• López C, Aramburu J, Galipienso L, Soler S, Nuez F, and Rubio L (2011) Evolutionary analysis of tomato Sw-5 resistance-breaking isolates of Tomato spotted wilt virus. Journal of General Virology 92(1): 210-215.
• McKinney JL, Tillman BL (2017) Spotted Wilt in Peanut as Impacted by Genotype Resistance, Planting Date, and Plant Population. Crop Science 57(1): 130-136.
• Moskova I, Sergiev I, Kirova E, and Dikova B (2021) Доклади на Българската академия на науките Effects of triacontanol on pepper plants infected with tomato spotted wilt virus (TSWV). Comptes rendus de l’Acade'mie bulgare des Sciences 74: 1091-1097.
• Mumford RA, Barker I, Wood KR (1994) The detection of tomato spotted wilt virus using the polymerase chain reaction. Journal of Virology Methods 46(3): 303-311.
• Ogada PA, Poehling HM (2015) Sex-Specific Influences of Frankliniella occidentalis (Western Flower Thrips) in the Transmission of Tomato spotted wilt virus (Tospovirus). Journal of Plant Disease and Protection 122: 264-274.
• Oğuz A, Ellialtioglu S, Çelik N, Kabaş A, Zengin S (2009) Bazi domates hatlarinin domates lekeli solgunluk virüsü (tswv=tomato spotted wilt virus)'ne karşi reaksiyonlarinin mekanik inokulasyon yöntemi ile belirlenmesi. Derim 26(1): 40-50.
• Pazarlar S, Gümüs M, Oztekin G (2013) The Effects of Tobacco mosaic virus Infection on Growth and Physiological Parameters in Some Pepper Varieties (Capsicum annuum L.). Notulae Botanicae Horti Agrobotanici 41: 427-433.
• Ramkat R, Wangai A, Ouma J, Rapando P, Lelgut D (2006) Effect of mechanical inoculation of Tomato spotted wilt tospovirus disease on disease severity and yield of greenhouse raised tomatoes. Asian Journal of Plants Sciences 5(10): 607-612.
• Salamon P, Szabó J (2016) Symptoms caused by Tomato spotted wilt virus (TSWV) in pepper (Capsicum spp.) and marker assisted selection of TSWV resistant pepper lines for hybrid constructions. Proceedings XVI. Eucarpia Capsicum and Eggplant Meeting 12: 69-75.
• Salaudeen M (2015) Growth and yield responses of some cowpea accessions to cucumber mosaic virus infection. Archives of Agronomy and Soil Science 62: 428-429.
• Şevik MA (2007) Domates lekeli solgunluk virüsü (TSWV)'nün Samsun ilinde domates üretim alanlarındaki yayılış durumunun ve bazı karakteristik özelliklerinin belirlenmesi. Doktora tezi. Tez no. 347165.
• Şevik MA, Cansız N (2021) The impact of turnip mosaic virus on physiological and morphological parameters of kale plants [Turnip mosaic virus’un yaprak lahana bitkilerinin fizyolojik ve morfolojik parametrelerine etkisi]. Gümüşhane Üniversitesi Fen Bilimleri Dergisi 11(3): 919-924.
• Shokoohi E (2023) First observation on morphological and molecular characters of Bitylenchus ventrosignatus (Tobar Jiménez, 1969) Siddiqi, 1986 isolated from tomato in Dalmada, South Africa. Biologia 78: 3599-3607.
• Svanella DL, Dulucq M, Candresse T, Gentit P, Foissac X (2000) Polyvalent detection of fruit tree tricho, capillo and foveaviruses by nested RT-PCR using degenerated and inosine containing primers (PDO RT-PCR). XVIII International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops-Top Fruit Diseases 550-552.
• Trojak GA, Laskowska D, Kursa K (2016) Morphological and chemical characteristics of doubled haploids of flue-cured tobacco combining resistance to Thielaviopsis basicola and TSWV. Breeding Science 66(2): 293-299.
• Turhan P, Korkmaz S (2006) Determination of Tomato spotted wilt virus using serological and biological methods in tomatoes grown in Çanakkale Province. Tarım Bilimleri Dergisi 12(2): 130-136.
• Usta M, Güller A, Demirel S, Korkmaz G, Kurt Z (2023) New insights into tomato spotted wilt orthotospovirus (TSWV) infections in Turkey: molecular detection, phylogenetic analysis, and in silico docking study. Notulae Botanicae Horti Agrobotanici 51: 1-22.
• Zhao D, Xia T, Zhou T, Zhao L, Zhu X, Gong B (2025) Inoculation method and disease evaluation of tomato chlorotic virus (ToCV) in Solanum lycopersicum. Vegetable Research 5(1).