Molecular identification, analysis, and sequencing of the coat protein gene of Tomato yellow leaf curl virus in tomato fields from selected regions of Iran

Document Type : Research Paper

Authors

Department of Plant Protection, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

Abstract

Tomato yellow leaf curl virus (TYLCV; Begomovirus coheni) is one of the most destructive plant viruses worldwide, causing severe yield losses in tomato fields. In this study, field surveys were conducted in tomato-growing areas of Isfahan, Yazd, and Varamin (central Iran) to detect and investigate the genetic diversity of TYLCV. Leaf samples showing virus-like symptoms were collected, and total DNA was extracted for molecular detection. PCR amplification using V1 gene-specific primers resulted in a ~670 bp fragment in 12 suspected samples, of which seven isolates were selected for sequencing. The obtained sequences were aligned with reference sequences from GenBank, and phylogenetic analyses were performed using the Neighbor-Joining method in MEGA11. The results revealed that selected Iranian isolates clustered into two distinct groups within clade I, along with isolates from other parts of the world. Haplotype network analysis further supported these groupings. Notably, phylogenetic placement and strain-level classification of the newly identified isolates showed that all seven Iranian isolates (PV579121–PV579127) clustered within the TYLCV-OM group, forming a distinct lineage. This close genetic relationship suggests a common evolutionary origin and supports the hypothesis that these isolates belong to a region-specific strain adapted to the environmental conditions of central Iran. Genetic diversity analyses revealed significant differences among virus populations from the surveyed regions. Furthermore, neutrality tests (Tajima’s D, Fu and Li’s D* and F*) and population genetic analyses provided evidence of purifying selection and recent population expansion. One significant recombination event was also detected using RDP4 software. Selection pressure analysis using FEL and MEME methods identified several codons under both positive and negative selection. Collectively, these findings provide a comprehensive picture of the genetic diversity and evolutionary dynamics of TYLCV in central Iran and offer valuable insights for developing effective regional disease management strategies.

Keywords


Bananej K, 2016. An analysis on the status of tomato yellow leaf curl disease. Applied Entomology & Phytopathology 84: 157–173.
Cohen S, & Harpaz I, 1964. Periodic, rather than continual acquisition of a new tomato virus by its vector, the tobacco whitefly (Bemisia tabaci gennadius) 1. Entomologia experimentalis et Applicata 7: 155–166.
Duffy S, & Holmes EC, 2008. Phylogenetic evidence for rapid rates of molecular evolution in the single-stranded DNA begomovirus tomato yellow leaf curl virus. Journal of Virology 82: 957–965.
Fiallo-Olivé E, Trenado HP, Louro D, & Navas-Castillo J, 2019. Recurrent speciation of a tomato yellow leaf curl geminivirus in Portugal by recombination. Scientific Reports 9: 1332.
Fidan H, Karacaoglu M, Koç G, & Caglar B, 2019. Tomato yellow leaf curl virus (TYLCV) strains and epidemiological role of Bemisia tabaci (Hemiptera: Aleyrodidae) biotypes on tomato agroecology in Turkey. Applied Ecology & Environmental Research 17.
Gambino G, Perrone I, & Gribaudo IJPA, 2008. A rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants  Phytochemical Analysis 19: 520–525.
García-Arenal F, & Zerbini FM, 2019. Life on the edge: geminiviruses at the interface between crops and wild plant hosts. Annual Review of Virology 6: 411–433.
García-Estrada RS, Diaz-Lara A, Aguilar-Molina VH, & Tovar-Pedraza JM, 2022. Viruses of economic impact on tomato crops in Mexico: From diagnosis to management—A review. Viruses 14: 1251.
Hajimorad M, Kheyr‐Pour A, Alavi V, Ahoonmanesh A, Bahar M, Rezaian M, & Gronenborn B, 1996. Identification of whitefly transmitted tomato yellow leaf curl geminivirus from Iran and a survey of its distribution with molecular probes. Plant Pathology 45: 418–425.
Hamilton C-L, Mazumdar-Leighton S, Amarakoon I, & Roye M, 2015. Tomato yellow leaf curl. In: Tennant P,  Fermin G (eds). Virus diseases of tropical and subtropical crops, CABI, Wallingford UK. Pp. 177–188.
Hanson PM, Bernacchi D, Green S, Tanksley SD, Muniyappa V, Padmaja AS, Chen H-M, Kuo G, Fang D, & Chen J-T, 2000. Mapping a wild tomato introgression associated with tomato yellow leaf curl virus resistance in a cultivated tomato line. Journal of the American Society for Horticultural Science 125: 15–20.
Kil E-J, Kim S, Lee Y-J, Byun H-S, Park J, Seo H, Kim C-S, Shim J-K, Lee J-H, & Kim J-K, 2016. Tomato yellow leaf curl virus (TYLCV-IL): a seed-transmissible geminivirus in tomatoes. Scientific Reports 6: 19013.
Lapidot M, & Polston JE, 2006. Resistance to tomato yellow leaf curl virus in tomato. Natural Resistance Mechanisms of Plants to Viruses 503–520.
Lefeuvre P, Martin DP, Harkins G, Lemey P, Gray AJ, Meredith S, Lakay F, Monjane A, Lett J-M, & Varsani A, 2010. The spread of tomato yellow leaf curl virus from the Middle East to the world. PLoS Pathogens 6: e1001164.
Li F, Qiao R, Yang X, Gong P, & Zhou X, 2022. Occurrence, distribution, and management of tomato yellow leaf curl virus in China. Phytopathology Research 4: 28.
Marchant WG, Gautam S, Hutton SF, & Srinivasan R, 2020. Tomato yellow leaf curl virus-resistant and-susceptible tomato genotypes similarly impact the virus population genetics. Frontiers in Plant Science 11: 599697.
Martin DP, Murrell B, Golden M, Khoosal A, & Muhire B, 2015. RDP4: Detection and analysis of recombination patterns in virus genomes. Virus Evolution 1(1): vev003.
Navot N, Pichersky E, Zeidan M, Zamir D, & Czosnek H, 1991. Tomato yellow leaf curl virus: a whitefly-transmitted geminivirus with a single genomic component. Virology 185: 151-161.
Ong SN, Taheri S, Othman RY, & Teo CH, 2020. Viral disease of tomato crops (Solanum lycopesicum L.): an overview. Journal of Plant Diseases & Protection 127: 725–739.
Rozas J, Ferrer-Mata A, Sánchez-DelBarrio JC, Guirao-Rico S, Librado P, Ramos-Onsins SE, Sánchez-Gracia A, 2017. DnaSP 6: DNA Sequence Polymorphism Analysis of Large Datasets. Molecular Biology & Evolution 34: 3299-3302.
Panno S, Caruso A, & Davino SJPD, 2019. First report of tomato brown rugose fruit virus on tomato crops in Italy.  Plant diseases 103: 1443–1443.
Pico B, Diez MJ, & Nuez F, 1999. Improved diagnostic techniques for tomato yellow leaf curl virus in tomato breeding programs. Plant Disease 83: 1006–1012.
Polston JE, & Lapidot M, 2007. Management of Tomato yellow leaf curl virus: US and Israel perspectives. Tomato Yellow Leaf Curl Virus Disease: management, molecular biology, breeding for resistance. Springer.
Prasad A, Sharma N, Hari-Gowthem G, Muthamilarasan M, & Prasad M, 2020. Tomato yellow leaf curl virus: impact, challenges, and management. Trends in Plant Science 25: 897–911.
Rubio L, Galipienso L, & Ferriol I, 2020. Detection of Plant Viruses and Disease Management: Relevance of Genetic Diversity and Evolution. Frontiers in Plant Science 11:1092. doi: 10.3389/fpls. 2020.0109 2.
Seymour GB, & Rose JK, 2022. Tomato molecular biology–special collection of papers for molecular horticulture. Molecular Horticulture 2: 21.
Shirazi M, Mozafari J, Rakhshandehroo F, & Shams-Bakhsh M, 2014. Genetic diversity, host range, and distribution of Tomato yellow leaf curl virus in Iran. Acta Virologica 58: 128–136.
Yan Z, Wolters A-MA, Navas-Castillo J, & Bai Y, 2021. The global dimension of tomato yellow leaf curl disease: Current status and breeding perspectives. Microorganisms 9: 740.
Yazdani-Khameneh S, & Golnaraghi A, 2017. The status of Begomoviruses in Iran. In: Saxena S, & Tiwari A K (eds). Begomoviruses: Occurrence and Management in Asia and Africa. Springer Nature, Singapore. Pp.  229–253.
Zhang L, Wu S, Zhao M, Ghanem H, Wu G, Li M, & Qing L, 2024. Molecular Identification and Genetic Diversity Analysis of Papaya Leaf Curl China Virus Infecting Ageratum conyzoides. The Plant Pathology Journal 40(5): 551.