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RT-RPA Assay Combined with a Lateral Flow Strip to Detect Soybean Mosaic Virus

  • Bong Geun Oh (Department of Agricultural Biology, Jeonbuk National University) ;
  • Ju-Yeon Yoon (Department of Plant Protection and Quarantine, Jeonbuk National University) ;
  • Ho-Jong Ju (Department of Agricultural Biology, Jeonbuk National University)
  • Received : 2024.02.07
  • Accepted : 2024.06.13
  • Published : 2024.08.01

Abstract

Soybean (Glycine max L.) is one of the most widely planted and used legumes in the world, being used for food, animal feed products, and industrial production. The soybean mosaic virus (SMV) is the most prevalent virus infecting soybean plants. This study developed a diagnostic method for the rapid and sensitive detection of SMV using a reverse transcription-recombinase polymerase amplification (RT-RPA) technique combined with a lateral flow strip (LFS). The RT-RPA and RT-RPA-LFS conditions to detect the SMV were optimized using the selected primer set that amplified part of the VPg protein gene. The optimized reaction temperature for the RT-RPA primer and RT-RPA-LFS primer used in this study was 38℃ for both, and the minimum reaction time was 10 min and 5 min, respectively. The RT-RPA-LFS was as sensitive as RT-PCR to detect SMV with 10 pg/µl of total RNA. The reliability of the developed RT-RPA-LFS assay was evaluated using leaves collected from soybean fields. The RT-RPA-LFS diagnostic method developed in this study will be useful as a diagnostic method that can quickly and precisely detect SMV in the epidemiological investigation of SMV, in the selection process of SMV-resistant varieties, on local farms with limited resources.

Keywords

Acknowledgement

This study was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Plant Virus and Industrialization in Response to Pests Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (Grant No: 120080-05). The authors would like to thank the Writing Center at Jeonbuk National University for its skilled proofreading service.

References

  1. Botelho, S. R., Martins, T. P., Duarte, M. F., Barbosa, A. V., Lau, D., Fernandes, F. R. and Sanches, M. M. 2016. Development of methodologies for virus detection in soybean and wheat seeds. MethodsX 3:62-68. https://doi.org/10.1016/j.mex.2016.01.005
  2. Cho, E.-K. 2007. Severity of cowpea mosaic virus and putkong disease monitoring and purification of cowpea mosaic virus. Res. Plant Dis. 13:30-33 (in Korean). https://doi.org/10.5423/RPD.2007.13.1.030
  3. Cho, E. K. and Chung, B. J. 1976. Studies on identification and classification of soybean virus diseases in Korea. Res. Plant Dis. 15:61-68.
  4. Crannell, Z., Castellanos-Gonzalez, A., Nair, G., Mejia, R., White, A. C. and Richards-Kortum, R. 2016. Multiplexed recombinase polymerase amplification assay to detect intestinal protozoa. Anal. Chem. 88:1610-1616. https://doi.org/10.1021/acs.analchem.5b03267
  5. Crannell, Z. A., Rohrman, B. and Richards-Kortum, R. 2014. Equipment-free incubation of recombinase polymerase amplification reactions using body heat. PLoS ONE 9:e112146.
  6. Del Rio, J. S., Steylaerts, T., Henry, O. Y. F., Bienstman, P., Stakenborg, T., Van Roy, W. and O'Sullivan, C. K. 2015. Realtime and label-free ring-resonator monitoring of solid-phase recombinase polymerase amplification. Biosens. Bioelectron. 73:130-137. https://doi.org/10.1016/j.bios.2015.05.063
  7. Domier, L. L., Hobbs, H. A., McCoppin, N. K., Bowen, C. R., Steinlage, T. A., Chang, S., Wang, Y. and Hartman, G. L. 2011. Multiple loci condition seed transmission of soybean mosaic virus (SMV) and SMV-induced seed coat mottling in soybean. Phytopathology 101:750-756. https://doi.org/10.1094/PHYTO-09-10-0239
  8. Domier, L. L., Steinlage, T. A., Hobbs, H. A., Wang, Y., HerreraRodriguez, G., Haudenshield, J. S., McCoppin, N. K. and Hartman, G. L. 2007. Similarities in seed and aphid transmission among soybean mosaic virus isolates. Plant Dis. 91:546-550. https://doi.org/10.1094/PDIS-91-5-0546
  9. Goodman, R. M. and Oard, J. H. 1980. Seed transmission and yield losses in tropical soybeans infected by soybean mosaic virus. Plant Dis. 64:913-914. https://doi.org/10.1094/PD-64-913
  10. Hajimorad, M. R., Domier, L. L., Tolin, S. A., Whitham, S. A. and Saghai Maroof, M. A. 2018. Soybean mosaic virus: a successful potyvirus with a wide distribution but restricted natural host range. Mol. Plant Pathol. 19:1563-1579. https://doi.org/10.1111/mpp.12644
  11. Higgins, M., Ravenhall, M., Ward, D., Phelan, J., Ibrahim, A., Forrest, M. S., Clark, T. G. and Campino, S. 2019. PrimedRPA: primer design for recombinase polymerase amplification assays. Bioinformatics 35:682-684. https://doi.org/10.1093/bioinformatics/bty701
  12. Irwin, M. E. and Goodman, R. M. 1981. Ecology and control of soybean mosaic virus. In: Plant diseases and vectors: ecology and epidemiology, eds. by K. Maramorosch and K. F. Harris, pp. 181-229. Academic Press, Cambridge, MA, USA.
  13. Jang, Y. W., Jo, Y., Cho, W. K., Choi, H., Yoon, Y. N., Lim, S. M., Lee, Y. H., Bae, J. Y. and Lee, B. C. 2018. First report of bean common mosaic necrosis virus infecting soybean in Korea. Plant Dis. 102:2051.
  14. Jayaram, C., Hill, J. H. and Miller, W. A. 1992. Complete nucleotide sequences of two soybean mosaic virus strains differentiated by response of soybean containing the Rsv resistance gene. J. Gen. Virol. 73:2067-2077. https://doi.org/10.1099/0022-1317-73-8-2067
  15. Jiang, T., Wang, Y., Jiao, W., Song, Y., Zhao, Q., Wang, T., Bi, J. and Shen, A. 2022. Recombinase polymerase amplification combined with real-time fluorescent probe for Mycoplasma pneumoniae detection. J. Clin. Med. 11:1780.
  16. Jo, Y., Yoon, Y. N., Jang, Y.-W., Choi, H., Lee, Y.-H., Kim, S.- M., Choi, S. Y., Lee, B. C. and Cho, W. K. 2020. Soybean viromes in the Republic of Korea revealed by RT-PCR and next-generation sequencing. Microorganisms 8:1777.
  17. Kil, E.-J., Park, J., Choi, H.-S., Kim, C.-S. and Lee, S. 2017. Seed transmission of tomato yellow leaf curl virus in white soybean (Glycine max). Plant Pathol. J. 33:424-428. https://doi.org/10.5423/PPJ.NT.02.2017.0043
  18. Kim, S.-M., Lee, J.-B., Lee, Y.-H., Choi, S.-H., Choi, H.-S., Park, J.-W., Lee, J.-S., Lee, G.-S., Moon, J.-K., Moon, J.-S., Lee, K.-W. and Lee, S.-H. 2006. First report of soybean dwarf virus on soybean (Glycine max) in Korea. Res. Plant Dis. 12:213-220 (in Korean). https://doi.org/10.5423/RPD.2006.12.3.213
  19. Kim, Y.-H., Kim, O.-S., Lee, B.-C., Roh, J.-H., Kim, M.-K., Im, D.-J., Hur, I.-B. and Lee, S.-C. 1999. Detection of soybean mosaic virus using RT-PCR. Korean J. Crop Sci. 44:253-255.
  20. Kunze, A., Dilcher, M., Abd El Wahed, A., Hufert, F., Niessner, R. and Seidel, M. 2016. On-chip isothermal nucleic acid amplification on flow-based chemiluminescence microarray analysis platform for the detection of viruses and bacteria. Anal. Chem. 88:898-905. https://doi.org/10.1021/acs.analchem.5b03540
  21. Lee, H.-J., Choi, S., Cho, I.-S., Yoon, J.-Y. and Jeong, R.-D. 2023. Development and application of a reverse transcription droplet digital PCR assay for detection and quantification of plantago asiatica mosaic virus. Crop Prot. 169:106255.
  22. Lee, H. J., Kim, M.-K., Lee, S. G., Choi, C. S., Choi, H.-S., Kwak, H. R., Choi, G. S. and Chun, C. 2015a. Physiological characteristics of melon plants showing leaf yellowing symptoms caused by CABYV infection. Hortic. Sci. Technol. 33:210-218 (in Korean). https://doi.org/10.7235/hort.2015.14149
  23. Lee, H. S., Lee, M. H. and Tochihara, H. 1980. An infectious virus isolated from soybeans. Korean J. Plant Prot. 9:175-179.
  24. Lee, H. T. 2022. Development of lateral flow reverse transcription recombinase polymerase amplification (LF RT-RPA) for detection of Cucumber mosaic virus (CMV) on Cnidium officinale in field. M.S. thesis. Jeonbuk National University, Jeonju, Korea.
  25. Lee, S. H., Choi, Y. M., Kim, J. S. and Chung, B. J. 1985. Identification of alfalfa mosaic virus from soybean. Korean J. Plant Pathol. 1:33-37 (in Korean).
  26. Lee, Y.-H., Bae, D.-H., Kim, B.-S., Yoon, Y.-N., Bae, S.-D. Kim, H.-J. Mainali, B. P., Park, I.-H., Lee, S.-H. and Kang, H.-W. 2015b. Detection of soybean mosaic virus by reverse transcription loop-mediated isothermal amplification. Res. Plant Dis. 21:351-320 (in Korean).
  27. Lee, Y. H., Lim, S. T., Yoon, Y. N., Jeon, M. G., Yun, H. T., Ko, J. M., Lee, S. H., Lee, K. W. and Baek, I. Y. 2012. Incidence of soybean viral diseases in Korea. Korea Soybean Digest 29:7-15.
  28. Lee, Y.-H., Yoon, Y.-N., Yun, H.-T., Baek, I. Y., Lim, S., Moon, J. S. and Lee, S.-H. 2015c. First report of bean common mosaic virus infecting soybean in South Korea. Plant Dis. 99:1189.
  29. Li, J., Macdonald, J. and von Stetten, F. 2019. Review: a comprehensive summary of a decade development of the recombinase polymerase amplification. Analyst 144:31-67. https://doi.org/10.1039/C8AN01621F
  30. Li, J., Zhong, Q., Shang, M.-Y., Li, M., Jiang, Y.-S., Zou, J.-J., Ma, S.-S., Huang, Q. and Lu, W.-P. 2022. Preliminary evaluation of rapid visual identification of Burkholderia pseudomallei using a newly developed lateral flow strip-based recombinase polymerase amplification (LF-RPA) system. Front. Cell. Infect. Microbiol. 11:804737.
  31. Libault, M. A. R. C., Thibivilliers, S., Bilgin, D. D., Radwan, O., Benitez, M., Clough, S. J. and Stacey, G. 2008. Identification of four soybean reference genes for gene expression normalization. Plant Genome 1:44-54. https://doi.org/10.3835/plantgenome2008.02.0091
  32. Lim, S., Lee, Y.-H., Igori, D., Zhao, F., Yoo, R. H., Lee, S.-H., Baek, I. Y. and Moon, J. S. 2014. First report of peanut mottle virus infecting soybean in South Korea. Plant Dis. 98:1285.
  33. Liu, D., Shen, H., Zhang, Y., Shen, D., Zhu, M., Song, Y., Zhu, Z. and Yang, C. 2021. A microfluidic-integrated lateral flow recombinase polymerase amplification (MI-IF-RPA) assay for rapid COVID-19 detection. Lab Chip 21:2019-2026. https://doi.org/10.1039/D0LC01222J
  34. Liu, J.-Z., Fang, Y. and Pang, H. 2016. The current status of the soybean-soybean mosaic virus (SMV) pathosystem. Front. Microbiol. 7:1906.
  35. Lobato, I. M. and O'Sullivan, C. K. 2018. Recombinase polymerase amplification: basics, applications and recent advances. Trends Anal. Chem. 98:19-35. https://doi.org/10.1016/j.trac.2017.10.015
  36. Lopez, M. M., Llop, P., Olmos, A., Marco-Noales, E., Cambra, M. and Bertolini, E. 2009. Are molecular tools solving the challenges posed by detection of plant pathogenic bacteria and viruses?. Curr. Issues Mol. Biol. 11:13-46.
  37. Maury, Y., Duby, C., Bossennec, J.-M. and Boudazin, G. 1986. Group analysis using ELISA: determination of the level of transmission of soybean mosaic virus in soybean seed. Agronomie 5:405-415. https://doi.org/10.1051/agro:19850504
  38. Mehetre, G. T., Leo, V. V., Singh, G., Sorokan, A., Maksimov, I., Yadav, M. K., Upadhyaya, K., Hashem, A., Alsaleh, A. N., Dawoud, T. M., Almaary, K. S. and Singh, B. P. 2021. Current developments and challenges in plant viral diagnostics: a systematic review. Viruses 13:412.
  39. Ministry of Agriculture, Food and Rural Affairs. 2022. Cultivation status of flowering plants in Korea 2021. URL http://lib.mafra.go.kr/Search/Detail/49887/ [16 May 2023].
  40. Nam, M., Kim, S. M., Domier, L. L., Koh, S., Moon, J. K., Choi, H. S., Kim, H. G., Moon, J. S. and Lee, S.-H. 2009. Nucleotide sequence and genomic organization of a newly identified member of the genus Carmovirus, soybean yellow mottle mosaic virus, from soybean. Arch. Virol. 154:1679-1684. https://doi.org/10.1007/s00705-009-0480-z
  41. Nam, M., Park, S.-J., Kim, Y.-J., Kim, J.-S., Park, C. Y., Lee, J.-S., Choi, H.-S., Kim, J.-S., Kim, H. G. and Lee, S.-H. 2012. First report of peanut stunt virus on Glycine max in Korea. Plant Pathol. J. 28:330. https://doi.org/10.5423/PPJ.DR.07.2011.0138
  42. Piepenburg, O., Williams, C. H., Stemple, D. L. and Armes, N. A. 2006. DNA detection using recombination proteins. PLoS Biol. 4:e204.
  43. Qi, Y., Yin, Q., Shao, Y., Li, S., Chen, H., Shen, W., Rao, J., Li, J., Li, X., Sun, Y., Lin, Y., Deng, Y., Zeng, W., Zheng, S., Liu, S. and Li, Y. 2018. Rapid and visual detection of Coxiella burnetii using recombinase polymerase amplification combined with lateral flow strips. Biomed Res. Int. 2018:6417354.
  44. Shin, J.-C., Kim, M.-K., Kwak, H.-R., Choi, H.-S., Kim, J.-S., Park, C.-Y. and Cha, B. J. 2014. First report of clover yellow vein virus on Glycine max in Korea. Plant Dis. 98:1283.
  45. Tamura, K., Stecher, G. and Kumar, S. 2021. MEGA11: molecular evolutionary genetics analysis version 11. Mol. Biol. Evol. 38:3022-3027. https://doi.org/10.1093/molbev/msab120
  46. Tolin, S. A. and Lacy, G. H. 2004. Viral, bacterial, and phytoplasmal diseases of soybean. In: Soybean: improvement, production, and uses, eds. by H. R. Boerma and J. E. Specht, pp. 765-819. American Society of Agronomy, Crop Science Society of America, Soil Science Society of America, Madison, WI, USA.
  47. Tsaloglou, M.-N., Nemiroski, A., Camci-Unal, G., Christodouleas, D. C., Murray, L. P., Connelly, J. T. and Whitesides, G. M. 2018. Handheld isothermal amplification and electrochemical detection of DNA in resource-limited settings. Anal. Biochem. 543:116-121. https://doi.org/10.1016/j.ab.2017.11.025
  48. Wang, J., Koo, K. M., Wee, E. J. H., Wang, Y. and Trau, M. 2017. A nanoplasmonic label-free surface-enhanced raman scattering strategy for non-invasive cancer genetic subtyping in patient samples. Nanoscale 9:3496-3503. https://doi.org/10.1039/C6NR09928A
  49. Yan, L., Zhou, J., Zheng, Y., Gamson, A. S., Roembke, B. T., Nakayama, S. and Sintim, H. O. 2014. Isothermal amplified detection of DNA and RNA. Mol. Biosyst. 10:970-1003. https://doi.org/10.1039/c3mb70304e
  50. Yonesaki, T. and Minagawa, T. 1985. T4 phage gene uvsX product catalyzes homologous DNA pairing. EMBO J. 4:3321-3327. https://doi.org/10.1002/j.1460-2075.1985.tb04083.x
  51. Yoon, J.-Y., Choi, G.-S., Cho, I.-S. and Choi, S.-K. 2014. Development of a novel sap extraction buffer for simultaneous detection of pepper viruses using VC/RT-PCR. J. Korean Soc. Int. Agric. 26:41-47 (in Korean). https://doi.org/10.12719/KSIA.2014.26.1.41
  52. Yoon, Y. N., Jo, Y., Cho, W. K., Choi, H., Jang, Y., Lee, Y. H., Bae, J. Y. and Lee, B. C. 2018. First report of tomato spotted wilt virus infecting soybean in Korea. Plant Dis. 102:461.
  53. Zhou, Y., Zheng, H. Y., Jiang, D. M., Liu, M., Zhang, W. and Yan, J. Y. 2022. A rapid detection of tomato yellow leaf curl virus using recombinase polymerase amplification-lateral flow dipstick assay. Lett. Appl. Microbiol. 74:640-646. https://doi.org/10.1111/lam.13611
  54. Zou, X., Dong, C., Ni, Y., Yuan, S. and Gao, Q.-H. 2022a. Rapid detection of strawberry mottle virus using reverse transcription recombinase polymerase amplification with lateral flow strip. J. Virol. Methods 307:114566.
  55. Zou, X., Yuan, S., Dong, C. and Gao, Q. 2022b. Development and evaluation of an isothermal recombinase polymerase amplification-lateral flow assay for rapid detection of strawberry vein banding virus in the field. Crop Prot. 158:105994.