Similar articles

Abstract: Pepino mosaic virus (PepMV) causes severe disease in tomato and other Solanaceous crops around globe. To effectively study and manage this viral disease, researchers need new, sensitive, and high-throughput approaches for viral detection. In this study, we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies (mAbs). A panel of highly specific and sensitive murine mAbs (15B2, 8H6, 23D11, 20D9, 3A6, and 8E3) could be produced through cell fusion, antibody selection, and cell cloning. Using the mAbs as the detection antibodies, we established double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), dot-ELISA, and tissue print-ELISA for detecting PepMV infection in tomato plants. Resulting data on sensitivity analysis assays showed that both DAS-ELISA and dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1 310 720 and 1:20 480 (weight/volume ratio (w/v), g/mL), respectively. Among the three methods developed, the tissue print-ELISA was found to be the most practical detection technique. Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing, demonstrating all three serological methods are reliable and effective for monitoring PepMV. Anti-PepMV mAbs and the newly developed DAS-ELISA, dot-ELISA, and tissue print-ELISA can benefit PepMV detection and field epidemiological study, and management of this viral disease, which is already widespread in tomato plants in Yunnan Province of China.

凤果花叶病毒高灵敏度血清学检测技术

[1]Aguilar JM, Hernández-Gallardo MD, Cenis JL, et al., 2002. Complete sequence of the Pepino mosaic virus RNA genome. Arch Virol, 147(10):2009-2015.

[2]Chen Z, Zhang MH, Zhou XP, et al., 2017. Development and detection application of monoclonal antibodies against Zucchini yellow mosaic virus. J Integr Agric, 16(1):115-124.

[3]Córdoba MC, Martínez-Priego L, Jordá C, 2004. New natural hosts of Pepino mosaic virus in Spain. Plant Dis, 88(8):906.

[4]Córdoba-Sellés MDC, García-Rández A, Alfaro-Fernández A, et al., 2007. Seed transmission of Pepino mosaic virus and efficacy of tomato seed disinfection treatments. Plant Dis, 91(10):1250-1254.

[5]Cotillon AC, Girard M, Ducouret S, 2002. Complete nucleotide sequence of the genomic RNA of a French isolate of Pepino mosaic virus (PepMV). Arch Virol, 147(11):2231-2238.

[6]French CJ, Bouthillier M, Bernardy M, et al., 2001. First report of Pepino mosaic virus in Canada and the United States. Plant Dis, 85(10):1121.

[7]Guo LQ, Wu JY, Chen R, et al., 2020. Monoclonal antibody-based serological detection of Rice stripe mosaic virus infection in rice plants or leafhoppers. Virol Sin, 35(2):227-234.

[8]Hanssen IM, Thomma BPHJ, 2010. Pepino mosaic virus: a successful pathogen that rapidly evolved from emerging to endemic in tomato crops. Mol Plant Pathol, 11(2):179-189.

[9]Hanssen IM, Paeleman A, Wittemans L, et al., 2008. Genetic characterization of Pepino mosaic virus isolates from Belgian greenhouse tomatoes reveals genetic recombination. Eur J Plant Pathol, 121(2):131-146.

[10]Hanssen IM, Mumford R, Blystad DR, et al., 2010. Seed transmission of Pepino mosaic virus in tomato. Eur J Plant Pathol, 126(2):145-152.

[11]Hasiów-Jaroszewska B, Borodynko N, 2013. Detection of Pepino mosaic virus isolates from tomato by one-step reverse transcription loop-mediated isothermal amplification. Arch Virol, 158(10):2153-2156.

[12]Hasiów-Jaroszewska B, Pospieszny H, Borodynko N, 2009. New necrotic isolates of Pepino mosaic virus representing the Ch2 genotype. J Phytopathol, 157(7-8):494-496.

[13]Huang DQ, Chen R, Wang YQ, et al., 2019. Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(4):343-354.

[14]Jones RAC, Koenig R, Lesemann DE, 1980. Pepino mosaic virus, a new potexvirus from pepino (Solanum muricatum). Ann Appl Biol, 94(1):61-68.

[15]Jordá C, Pérez AL, Martínez Culebras PV, et al., 2001. First report of Pepino mosaic virus on natural hosts. Plant Dis, 85(12):1292.

[16]Li FF, Zhang CW, Li YZ, et al., 2018. Beclin1 restricts RNA virus infection in plants through suppression and degradation of the viral polymerase. Nat Commun, 9:1268.

[17]Li YM, Fan PH, Zhou SS, et al., 2017. Loop-mediated isothermal amplification (LAMP):a novel rapid detection platform for pathogens. Microb Pathog, 107:54-61.

[18]Ling KS, 2008. Pepino mosaic virus on tomato seed: virus location and mechanical transmission. Plant Dis, 92(12):1701-1705.

[19]Ling KS, Wechter WP, Jordan R, 2007. Development of a one-step immunocapture real-time TaqMan RT-PCR assay for the broad spectrum detection of Pepino mosaic virus. J Virol Methods, 144(1-2):65-72.

[20]Ling KS, Wintermantel WM, Bledsoe M, 2008. Genetic composition of Pepino mosaic virus population in North American greenhouse tomatoes. Plant Dis, 92(12):1683-1688.

[21]Liu H, Song XJ, Ni YQ, et al., 2014. Highly sensitive and specific monoclonal antibody-based serological methods for Rice ragged stunt virus detection in rice plants and rice brown planthopper vectors. J Integr Agric, 13(9):1943-1951.

[22]Liu Z, Sunzhu YJ, Zhou XP, et al., 2017. Monoclonal antibody-based serological detection of Citrus yellow vein clearing virus in citrus groves. J Integr Agric, 16(4):884-891.

[23]Mansilla C, Sánchez F, Ponz F, 2003. The diagnosis of the tomato variant of pepino mosaic virus: an IC-RT-PCR approach. Eur J Plant Pathol, 109(2):139-146.

[24]Maroon-Lango CJ, Guaragna MA, Jordan RL, et al., 2005. Two unique US isolates of Pepino mosaic virus from a limited source of pooled tomato tissue are distinct from a third (European-like) US isolate. Arch Virol, 150(6):1187-1201.

[25]Pagán I, Córdoba-Sellés MDC, Martínez-Priego L, et al., 2006. Genetic structure of the population of Pepino mosaic virus infecting tomato crops in Spain. Phytopathology, 96(3):274-279.

[26]Roggero P, Masenga V, Lenzi R, et al., 2001. First report of Pepino mosaic virus in tomato in Italy. Plant Pathol, 50(6):798-798.

[27]Salomone A, Roggero P, 2002. Host range, seed transmission and detection by ELISA and lateral flow of an Italian isolate of Pepino mosaic virus. J Plant Pathol, 84(1):65-68.

[28]Shang HL, Xie Y, Zhou XP, et al., 2011. Monoclonal antibody-based serological methods for detection of Cucumber green mottle mosaic virus. Virol J, 8:228.

[29]Shipp JL, Buitenhuis R, Stobbs L, et al., 2008. Vectoring of Pepino mosaic virus by bumble-bees in tomato greenhouses. Ann Appl Biol, 153(2):149-155.

[30]Song G, Wu JY, Xie Y, et al., 2017. Monoclonal antibody-based serological assays for detection of Potato virus S in potato plants. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 18(12):1075-1082.

[31]Spence NJ, Basham J, Mumford RA, et al., 2006. Effect of Pepino mosaic virus on the yield and quality of glasshouse-grown tomatoes in the UK. Plant Pathol, 55(5):595-606.

[32]van der Vlugt RAA, Stijger CCMM, Verhoeven JTJ, et al., 2000. First report of Pepino mosaic virus on tomato. Plant Dis, 84(1):303.

[33]Verhoeven JTJ, van der Vlugt R, Roenhorst JW, 2003. High similarity between tomato isolates of Pepino mosaic virus suggests a common origin. Eur J Plant Pathol, 109(5):419-425.

[34]Wu JX, Meng CM, Shang HL, et al., 2011. Monoclonal antibody-based triple antibody sandwich-enzyme-linked immunosorbent assay and immunocapture reverse transcription-polymerase chain reaction for Odontoglossum ringspot virus detection. J Virol Methods, 171(1):40-45.

[35]Wu JX, Ni YQ, Liu H, et al., 2014. Monoclonal antibody-based serological assays and immunocapture-RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors. J Virol Methods, 195:134-140.

[36]Zhang MH, Chen R, Zhou XP, et al., 2018. Monoclonal antibody-based serological detection methods for wheat dwarf virus. Virol Sin, 33(2):173-180.

[37]Zhang Y, Gao YL, He WQ, et al., 2020. Monoclonal antibody-based serological detection of potato virus M in potato plants and tubers. J Integr Agric, 19(5):1283-1291.

[38]Zhou XP, Chen JH, Li DB, et al., 1994. A method for high yield purification of potyviruse. Microbiology China, 21(3):184-186 (in Chinese).