The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

Rapid and Sensitive Detection of Lettuce Necrotic Yellows Virus and Cucumber Mosaic Virus Infecting Lettuce (Lactuca sativa L.) by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Zhang, Yubao (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ;
  • Xie, Zhongkui (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ;
  • Fletcher, John D (The New Zealand Institute for Plant and Food Research) ;
  • Wang, Yajun (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ;
  • Wang, Ruoyu (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ;
  • Guo, Zhihong (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences) ;
  • He, Yuhui (Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences)
  • Received : 2019.12.16
  • Accepted : 2020.01.10
  • Published : 2020.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg2+ at 65℃ for LNYV and 60℃ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.

Keywords

References

  1. Bhat, A. I., Siljo, A. and Deeshma, K. P. 2013. Rapid detection of Piper yellow mottle virus and Cucumber mosaic virus infecting black pepper (Piper nigrum) by loop-mediated isothermal amplification (LAMP). J. Virol. Methods 193:190-196. https://doi.org/10.1016/j.jviromet.2013.06.012
  2. Blancard, D., Lot, H. and Maisonneuve, B. 2006. A color atlas of diseases of lettuce and related salad crops: observation, biology and control. Academic Press, Boston, MA, USA. 375 pp.
  3. Bruckart, W. L. and Lorbeer, J. W. 1975. Recent occurrences of cucumber mosaic, lettuce mosaic and broad bean wilt viruses in lettuce and celery fields in New York. Plant Dis. Rep. 59:203-206.
  4. Callaghan, B. and Dietzgen, R. G. 2005. Nucleocapsid gene variability reveals two subgroups of Lettuce necrotic yellows virus. Arch. Virol. 150:1661-1667. https://doi.org/10.1007/s00705-005-0528-7
  5. Deyong, Z., Willingmann, P., Heinze, C., Adam, G., Pfunder, M., Frey, B. and Frey, J. E. 2005. Differentiation of cucumber mosaic virus isolates by hybridization to oligonucleotides in a microarray format. J. Virol. Methods 123:101-108. https://doi.org/10.1016/j.jviromet.2004.09.021
  6. Dietzgen, R. G. and Francki, R. I. B. 1988. Analysis of lettuce necrotic yellows virus structural proteins with monoclonal antibodies and concanavalin A. Virology 166:486-494. https://doi.org/10.1016/0042-6822(88)90519-3
  7. El-Borollosy, A. M. and Waziri, H. M. A. 2013. Molecular characterization of a cucumber mosaic cucumovirus isolated from lettuce in Egypt. Ann. Agric. Sci. 58:105-109. https://doi.org/10.1016/j.aoas.2013.01.014
  8. Fletcher, J. D., France, C. M. and Butler, R. C. 2005. Virus surveys of lettuce crops and management of lettuce big-vein disease in New Zealand. N. Z. Plant Prot. 58:239-244.
  9. Fletcher, J. D., Walker, M., Davidson, M., Paull, S. and Palmer, A. 2017. Outdoor lettuce virus disease project 2016-2018 Year 1 report. Plant & Food Research SPTS No. 14591. URL https://www.freshvegetables.co.nz/assets/Lettuce-virus-disease-project-Report-John-Flectcher-PFR-2016-2018-Year-1-FINAL.pdf [10 May 2019].
  10. Fry, P. R., Close. R. C., Procter, C. H. and Sunde, R. 1973. Lettuce necrotic yellows virus in New Zealand. N. Z. J. Agric. Res. 16:143-146. https://doi.org/10.1080/00288233.1973.10421173
  11. Gambley, C. F., Geering, A. D. W. and Thomas, J. E. 2009. Development of an immunomagnetic capture-reverse transcriptase-PCR assay for three pineapple ampeloviruses. J. Virol. Methods 155:187-192. https://doi.org/10.1016/j.jviromet.2008.10.006
  12. Hardinge, P., Kiddle, G., Tisi, L. and Murray, J. A. H. 2018. Optimised LAMP allows single copy detection of 35Sp and NOSt in transgenic maize using bioluminescent assay in real time (BART). Sci. Rep. 8:17590. https://doi.org/10.1038/s41598-018-36207-4
  13. He, X., Xue, F., Xu, S. and Wang, W. 2016. Rapid and sensitive detection of Lily symptomless virus by reverse transcription loop-mediated isothermal amplification. J. Virol. Methods 238:38-41. https://doi.org/10.1016/j.jviromet.2016.10.003
  14. Higgins, C. M., Chang, W.-L., Khan, S., Tang, J., Elliott, C. and Dietzgen, R. G. 2016. Diversity and evolutionary history of lettuce necrotic yellows virus in Australia and New Zealand. Arch. Virol.161:269-277. https://doi.org/10.1007/s00705-015-2626-5
  15. Jackson, A. O., Dietzgen, R. G., Goodin, M. M., Bragg, J. N. and Deng, M. 2005. Biology of plant rhabdoviruses. Annu. Rev. Phytopathol. 43:623-660. https://doi.org/10.1146/annurev.phyto.43.011205.141136
  16. Koenig, R. 1981. Indirect ELISA methods for broad specificity detection of plant viruses. J. Gen. Virol. 55:53-62. https://doi.org/10.1099/0022-1317-55-1-53
  17. Krause-Sakate, R., Mello, R. N., Pavan, M. A., Zambolim, E. M., Carvalho, M. G., Le Gall, O. and Zerbini, F. M. 2001. Molecular characterization of two Brazilian isolates of Lettuce mosaic virus with distinct biological properties. Fitopatol. Bras. 26:153-157. https://doi.org/10.1590/S0100-41582001000200006
  18. Liang, Q.-L., Wei, L.-X. and Xu, B.-L. 2008. Study on the host range of viruses infecting ornamental lily. J. Gansu Agric. Univ. 43:94-96 (in Chinese).
  19. Liu, W.-H., Hong, J., Chen, J.-S. and Ye, M.-Q. 2004. Cloning and phylogenetic analysis of CP gene for two CMV isolates infecting Lilium cv. Oriental Hybrids. J. Agric. Biotechnol. 12:442-445 (in Chinese). https://doi.org/10.3969/j.issn.1674-7968.2004.04.018
  20. Liu, Y., Wang, Z., Qian, Y., Mu, J., Shen, L., Wang, F. and Yang, J. 2010. Rapid detection of tobacco mosaic virus using the reverse transcription loop-mediated isothermal amplification method. Arch. Virol. 155:1681-1685. https://doi.org/10.1007/s00705-010-0746-5
  21. Ma, C., Wang, Y., Zhang, P. and Shi, C. 2017. Accelerated isothermal nucleic acid amplification in betaine-free reaction. Anal. Biochem. 530:1-4. https://doi.org/10.1016/j.ab.2017.04.017
  22. Masuta, C., Seshimo, Y., Mukohara, M., Jung, H. J., Ueda, S., Ryu, K. H. and Choi, J. K. 2002. Evolutionary characterization of two lily isolates of Cucumber mosaic virus isolated in Japan and Korea. J. Gen. Plant Pathol. 68:163-168. https://doi.org/10.1007/PL00013070
  23. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N. and Hase, T. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28:e63. https://doi.org/10.1093/nar/28.12.e63
  24. Palukaitis, P., Roossinck, M. J., Dietzgen, R. G. and Francki, R. I. B. 1992. Cucumber mosaic virus. Adv. Virus Res. 41:281-348. https://doi.org/10.1016/S0065-3527(08)60039-1
  25. Parida, M., Sannarangaiah, S., Dash, P. K., Rao, P. V. L. and Morita, K. 2008. Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev. Med. Virol. 18:407-421. https://doi.org/10.1002/rmv.593
  26. Ragozzino, A., Alioto, D., Iengo, C. and Iego, C. 1989. The yellowing virus and mycoplasma diseases of lettuce in Campania and Latium regions. Riv. Patol. Veget. 25:15-19.
  27. Ravindran, A., Levy, J., Pierson, E. and Gross, D. C. 2012. Development of a loop-mediated isothermal amplification procedure as a sensitive and rapid method for detection of 'Candidatus Liberibacter solanacearum' in potatoes and psyllids. Phytopathology 102:899-907. https://doi.org/10.1094/PHYTO-03-12-0055-R
  28. Roossinck, M. J. 1999. Cucumoviruses (Bromoviridae) general features. In: Encyclopedia of virology, 2nd ed., eds. by L. Granoof and R. G. Webster, pp. 315-320. Academic Press, San Diego, CA, USA.
  29. Rubio-Huertos, M. and Garcia-Hidalgo, F. 1982. A rhabdovirus resembling lettuce necrotic yellows from lettuce in Spain. Phytopathol. Z. 103:232-238. https://doi.org/10.1111/j.1439-0434.1982.tb01747.x
  30. Stubbs, L. L. and Grogan, R. G. 1963. Necrotic yellows: a newly recognized virus disease of lettuce. Aust. J. Agric. Res. 14:439-459. https://doi.org/10.1071/AR9630439
  31. Tordo, N., Benmansour, A., Calisher, C., Dietzgen, R. G., Fang, R.-X., Jackson, A. O., Kurath, G., Nadin-Davis, S., Tesh, R. B. and Walker, P. J. 2005. Family Rhabdoviridae. In: Virus taxonomy: eighth report of the International Committee on the Taxonomy of Viruses, 2nd ed., eds. by C. M. Fauqent, M. A. Mayo, J. Maniloff, U. Desselberger and L. A. Ball, pp. 623-644. Elsevier Academic Press, San Diego, CA, USA.
  32. Viswanathan, R., Ganesh Kumar, V., Karuppaiah, R., Scindiya, M. and Chinnaraja, C. 2013. Development of duplex-immunocapture (Duplex-IC) RT-PCR for the detection of sugarcane streak mosaic virus and sugarcane mosaic virus in sugarcane. Sugar Tech. 15:399-405. https://doi.org/10.1007/s12355-013-0216-y
  33. Wikipedia: the free encyclopedia. 2019. Lettuce. URL https://en.wikipedia.org/w/index.php?title=Lettuce&oldid=894244593 [10 May 2019].
  34. Zhang, Y., Wang, Y., Xie, Z., Yang, G., Guo, Z. and Wang, L. 2017. Simultaneous detection of three lily viruses using Triplex IC-RT-PCR. J. Virol. Methods 249:69-75. https://doi.org/10.1016/j.jviromet.2017.08.018
  35. Zhao, K., Liu, Y. and Wang, X. 2010. Reverse transcription loopmediated isothermal amplification of DNA for detection of Barley yellow dwarf viruses in China. J. Virol. Methods 169:211-214. https://doi.org/10.1016/j.jviromet.2010.06.020
  36. Zhao, B., Yang, D., Zhang, Y., Xu, Y., Zhao, X., Liang, J., Fan, X., Du, Y., Zhu, Z., Shi, B., Zhang, Q., Zhang, X., Cai, Y. and Zhao, K. 2018. Rapid visual detection of lily mottle virus using a loop‑mediated isothermal amplification method. Arch. Virol.163:545-548. https://doi.org/10.1007/s00705-017-3618-4