Development and validation of diagnosis methods for wheat streak mosaic virus (WSMV)

Wheat streak mosaic virus, or Tritimovirus tritici; acronym – WSMV, is a harmful pest of wheat, corn, oats, millet, rye, sorghum, triticale, barley and numerous species of perennial cereal grasses. It can lead to wheat yield losses of up to 100%. WSMV is spread by contact, seed and the wheat curl mite Aceria tosichella Keifer. 16 countries importing Russian grain products include the absence of this virus as a prerequisite in the phytosanitary requirements.

To develop methodological guidelines for the detection and identification of WSMV, three ELISA test systems from leading manufacturers, four RT-PCR test systems in “real time” (real-time RT-PCR) and 8 pairs of primers for classical RT-PCR were tested. A sufficiently high level of specificity to the target object was established for the ELISA test systems for WSMV from Agdia, DSMZ and Loewe. The coincidence of the results of serological tests with the test system of Agdia and the results of molecular tests was 90.5–100%, which allows us to recommend this test system for screening tests for the presence of WSMV. It was established that reagent kits for RT-PCR to WSMV from Russian manufacturers can be recommended for screening tests for the presence of WSMV during phytosanitary monitoring of crops and for the analysis of grain seeds intended for use within the Russian Federation. For the analysis of grain products intended for export, an alternative to the ELISA method may be a test in the format of a 1-stage real-time RT-PCR with primers and probe WSMV F1/WSMV R1/WSMV P. For conducting confirmatory tests for the presence of WSMV, a protocol of 1-stage classical RT-PCR with primers PMB-70/PMB-71 has been developed, which is not inferior in sensitivity to tests using the PCR method in “real time”. The possibility of using primers WSMV-F/WSMV-R and WSMVL2/WSMVR2 as confirmatory tests has been established. To study the genetic characteristics of the detected isolates, it is advisable to use the primers WSMV-CP-AM-F2/WSMV-CP-AM-R and WSMV-CP-Czech-F/WSMV-CP-Czech-R as additional tests, which amplify the complete nucleotide sequences of the coat protein genes of the isolates of the WSMV-D and WSMV-B strains, respectively.

1. Ablova I.B., Bespalova L.A., Mokrousov V.V., Ablova O.S., Boyko A.P. Viral diseases of cereal crops in Krasnodar Krai [Virusnyye bolezni zernovykh kultur v Krasnodarskom kraye] // Plant protection and quarantine. 2012; 6: 14–17. (In Russ.)

2. Bogoutdinov D.Z., Kastalyeva T.B., Girsova N.V. Viral diseases of grain crops in Samara Oblast [Virusnyye zabolevaniya zernovykh kultur v Samarskoy oblasti] // Bulletin of the Orenburg State University. 2017; 4 (204): P. 46–52. (In Russ.)

3. Vlasov Yu.I., Larina E.I. Agricultural virology [Selskokhozyaystvennaya virusologiya] // M., Kolos. 1982; 240 p. (In Russ.)

4. Glinushkin A.P. Phytopathogenic complex of wheat and control measures [Fitopatogennyy kompleks pshenitsy i mery borby s nim] // Abstract of diss… doctor of agricultural sciences. – M., 2013; 38 p. (In Russ.)

5. Demyanenko F.P., Budzanivska I.Kh., Polischuk V.P., Kholonenko O.L. Using mathematical models to assess the prevalence of phytoviruses in some regions of Ukraine [Ispolzovaniye matematicheskikh modeley dlya otsenki rasprostranennosti fitovirusov v nekotorykh regionakh Ukrainy] // Abstracts of the 2nd international conference “Bioresources and viruses”. Kyiv, 1998; 4-5. (In Russ.)

6. Zhivaeva T.S., Prikhodko Yu.N., Shneyder Yu.A., Lozovaya E.N., Pruchkina M.A., Selyavkin S.N., Khorina N.A., Karimova E.V. Seromonitoring of viruses in wheat crops on the territory of the Russian Federation [Seromonitoring virusov v posevakh pshenitsy na territorii Rossiyskoy Federatsii] // V All-Russian Congress on Plant Protection. Collection of abstracts of reports. SPb.: FGBNU VIZR, 2024; 44. (In Russ.)

7. Prikhodko Yu. N., Zhivaeva T. S., Lozovaya E. N., Shneyder Yu. A., Pruchkina M. A., Bashkirova I. G., Karimova E. V., Selyavkin S. N., Kasatkin D. G., Komarov D. A., Kobzar V. F., Ablova I. B., Puzyrnaya O. Yu., Kerimov R. V. Species composition and prevalence of wheat viruses in the Russian Federation [Vidovoy sostav i rasprostranennost virusov pshenitsy v Rossiyskoy Federatsii] // Plant Health and Quarantine. 2024; 4S (20B): 71-72.https://doi.org/10.69536/FKR.2024.38.66.001. (In Russ.)

8. Prikhodko Yu.N., Zhivaeva T.S., Lozovaya E.N., Selyavkin S.N., Shneyder Yu.A., Pruchkina M.A., Karimova E.V. Seromonitoring of wheat viruses (2021–2023) [Seromonitoring virusov pshenitsy (2021–2023 gg.)] // Plant Health and Quarantine. 2024; S2-1 (18): 44. (In Russ.)

9. Razvyazkina G.M., Kalkova E.A., Belyanchikova Yu.V. Wheat stripe mosaic virus [Virus polosatoy mozaiki pshenitsy] // Plant protection from pests and diseases. 1963; 9: 54-55. (In Russ.)

10. Bennypaul H., Abdullahi I., Harding M.W., Aboukhaddour R. First detection of European isolates of Wheat streak mosaic virus in Canada// Plant Disease. – 2019. – Vol.103. – P. 1442. https://doi.org/10.1094/PDIS-12-18-2193-PDN.

11. Coutts B.A., Banovic M., Kehoe M.A., Severtson D.L., Jones R.A.C. Epidemiology of Wheat streak mosaic virus in wheat in a Mediterranean-type environment// Eur. J. Plant Pathol. – 2014 a. – Vol. 140. – P. 797-813. DOI:10.1007/s10658-014-0510-x.

12. Coutts B.A., Hammond N.E.B., Kehoe M.A., Jones, R.A.C. Finding Wheat streak mosaic virus in southwest Australia// Aust. J. Agric. Res. – 2008 a. – Vol. 59. – P. 836–843. https://doi.org/10.1071/AR08034.

13. Deb M., Anderson J.M. Development of a multiplexed PCR detection method for Barley and Cereal yellow dwarf viruses, Wheat spindle streak virus, Wheat streak mosaic virus and Soil-borne wheat mosaic virus// J. Virol. Methods. – 2008. – Vol. 148. – P. 17–24. DOI: 10.1016/j.jviromet.2007.10.015.

14. Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl. Acids. Symp. Ser.41: 95-98.

15. Inoue-Nagata A.K., Jordan R., Kreuze J., Li F., López-Moya J.J., Mäkinen K., Ohshima K., Wylie S.J. ICTV Virus Taxonomy Profile: Potyviridae// Journal of General Virology. – 2022. - 103:001738.

16. Jones A.C. Global plant virus disease pandemics and epidemics// Plants. – 2021, 10, 233. – https://doi.org/10.3390/plants10020233.

17. Jones R. A.C. Developing a sensitive dry seed test to detect seed-borne viruses in bulk seed samples of wheat// Project DAW00141, Department of Agriculture and Food Western Australia. – (DAFWA). – 2008. – 56 s.

18. Jones R.A.C., Vazquez-Iglesias I., Hajizadeh M., McGreig S., Gibbs A.J. Phylogenetics and evolution of Wheat streak mosaic virus: Its global origin and the source of the Australian epidemic// Plant Pathology. – 2022. – Vol.71. – P.1660-1673. https://doi.org/10.1111/ppa.13609.

19. Kúdela O., Kúdelová M., Nováková S., Glasa M. First report of Wheat streak mosaic virus in Slovakia// Plant Disease. – 2008. – Vol. 92, No. 9. – P. 1365. DOI: 10.1094/PDIS-92-9-1365C.

20. Lanoiselet V.M., Hind-Lanoiselet T.L., Murray G.M. Studies on the seed transmission of Wheat streak mosaic virus// Australasian Plant Pathology. – 2008. – Vol.37. – P. 584-588. https://doi.org/10.1071/AP08059.

21. Lee S., Kang E.-H., Chu Y.M., Shin Y.-G., Ahn T.-Y. Development of PCR diagnosis system for plant quarantine seed-borne Wheat streak mosaic virus// Korean Journal of Microbiology. – 2013. – Vol. 49, No. 2. – P.112–117. https://doi.org/10.7845/kjm.2013.3013.

22. Masumi M., Rastegar M., Zare A., Lesemann D.E., Ebrahim-Nesbat F., Izadpanah K. Wheat eqlid mosaic virus: a novel Potyviridae in Iran// Parasitica. – 2005. – Vol. Vol 61 (1). – P.101–104.

23. Mishchenko L.T., Dunich S.A., Mishchenko I.A., Petrenkova V.P., Mukha T.I. Monitoring of economically important wheat viruses under weather conditions change in Ukraine and investigation of seed transmission of Wheat streak mosaic virus// Bulgarian Journal of Agricultural Science. – 2018. – Vol. 24 (No 4). – P. 660–669.

24. Navia D., de Mendonça R.S., Skoracka A., Szydo W., Knihinicki D., Hein G.L., da Pereira P.R.V., Truol G., Lau D. Wheat curl mite, Aceria tosichella, and transmitted viruses: an expanding pest complex affecting cereal crops// Experimental and Applied Acarology. – 2013. – Vol. 59 (1/2). – P. 95–143. https://doi.org/10.1007/s10493-012-9633-y.

25. Price J.A., Smitha J., Simmons A., Fellers J., Rusha C.M. Multiplex real-time RT-PCR for detection of Wheat streak mosaic virus and Tritcum mosaic virus// Journal of Virological Methods. – 2010. – Vol. 165. – P. 198–201. DOI: 10.1016/j.jviromet.2010.01.019.

26. Rabenstein F., Seifers D.L., Schubert J., French R., Stenger D.C. Phylogenetic relationships, strain diversity and biogeography of tritimoviruses// Journal of General Virology. - 2002. – Vol.83 (4). – P.895-906. DOI: 10.1099/0022-1317-83-4-895.

27. Redila C.D., Prakash V., Nouri S. Metagenomics analysis of the wheat virome identifies novel plant and fungal-associated viral sequences// Viruses. – 2021. - 13, 2457. https://doi.org/10.3390/v13122457.

28. Rogers S.M., Payton M., Allen R.W., Melcher U., Carver J., Fletcher J. Method: a single nucleotide polymorphism genotyping method for Wheat streak mosaic virus// Genetics 2012, 3:10. - http://www.investigativegenetics.com/content/3/1/10.

29. Tamura K., Stecher G., and Kumar S. (2021) MEGA11: Molecular Evolutionary Genetics Analysis version 11. Molecular Biology and Evolution 38: 3022-3027.

30. Tatineni S., Graybosch R.A., Hein G.L., Wegulo S.N., French R. Wheat cultivar-specific disease synergism and alteration of virus accumulation during co-infection with Wheat streak mosaic virus and Triticum mosaic virus// Phytopathology. – 2010. – Vol.100 (3). – P.230-238. DOI: 10.1094/PHYTO-100-3-0230.

31. Tatineni S., Hein G.L. Tritimoviruses and Rymoviruses (Potyviridae)// Taxonomy – International Committee on Taxonomy of Viruses (ICTV). – 2020. - https://talk.ictvonline.org/taxonomy/.

32. CABI, 2025. Wheat streak mosaic virus (wheat streak) datasheet // http://www.cabi.org/isc/datasheet/56858 (last accessed – 11.03.2025 г).

33. https://blast.ncbi.nlm.nih.gov/Blast.cgi (last accessed: 15.01.2025 г.).