Mikrobiol. Z. 2018; 80(1):77-88. Ukrainian.
doi: https://doi.org/10.15407/microbiolj80.01.077

The Diagnosis of Beet Necrotic Yellow Vein Virus Circulating in Ukraine

Hrynchuk K.1, Antipov I.1, Kyrychenko A.2, Kraeva H.2, Likhanov A.3

1National University of Life and Environmental Sciences of Ukraine
15 Heroyiv Oborony Str., Kyiv, 03041, Ukraine

2Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

3Institute of Evolutionary Ecology, NAS of Ukraine
37 Akad. Lebedeva Str., Kyiv, 03143, Ukraine

Aim. This study was conducted to determine the prevalence of Beet necrotic yellow vein virus (BNYVV) circulating in agrocenoses of Ukraine and to develop some methods for detecting and identifying thе virus. Methods. The classical virology and molecular biological methods were used for the diagnosis and identification of viral pathogens, in particular, electron microscopy, plant-indicator method, polymerase chain reaction (PCR). The BNYVV was purified and all Koch’s postulates to improve infectious nature of the disease were fulfilled. To investigate the plant cell responses to the virus infections we performed histochemical analysis. Results. The study contains results on various methods used for the diagnosis of BNYVV. Polyclonal rabbit sera, obtained as a result of virus immunization contained high-titre antibodies. Specific antiviral antibodies can be used for a serological diagnostic system development. The presence of BNYVV has also been confrmed by electron microscopy and mechanical inoculation of the indicator plant. It was established significant decreasing of the cell-wall-bound peroxidase concentration in phloem cells of conducting beams of sugar beet infected with BNYVV. At the same time, the accumulation of free isoenzymes in protoplasts was observed. Such a redistribution of peroxidase molecules in cell compartments is accompanied by a significant increase in their activity. Conclusions. The survey of agrocenoses in Cherkassy region showed sugar beets are infected with both Beet necrotic yellow vein virus and Beet mosaic virus. The presence of a mixed infection in plants was confrmed by PCR. Indicator plants using in bio-test detection of the virus produced typical BNYVV symptoms. It was shown the possibility of rapid BNYVV diagnosis by the histochemical method. It is recommended to examine the virus presence both in leaves and in root crops of sugar beet plants.

Keywords: beet necrotic yellow vein virus, beet mosaic virus, identifcation of BNYVV, sugar beet, polymerase chain reaction.

Full text (PDF, in Ukrainian)

  1. Andrusyk I, Antipov I, Bogach E. [PCR-diagnostics and identifcation of beet mosaic virus.] Biological Resources and Nature Management. 2014; 6(5-6):11-13. Ukrainian.
  2. Andrusyk I, Antipov I, Kyrychenko A. [PCR diagnosis and identifcation of sugar beet yellow virus.] Visnyk of the Sumy National University. 2013; 11(26):42–46. Ukrainian.
  3. Antibodies. Methods. In: Catty D. editor. Moscow: Mir, 1991; 2. 384 p. Russian.
  4. Brown F. The classifcation and nomenclature of viruses. Summary of results of meetings of International Committee on Taxonomy of Viruses in Edmonton, Canada. Intervirology. 1989; 30(4):181-86. https://doi.org/10.1159/000150091
  5. Fauquet MC, Mayo MA, Manilof J et al. Virus Taxonomy: VIIIth Report of the International Committee on Taxonomy of Viruses. USA: Elsevier, 2005; 1562.
  6. Finkenstadt VL. A review on the complete utilization of the sugar beet. Sugar Tech. 2013; 16(4):339-46. https://doi.org/10.1007/s12355-013-0285-y
  7. Furst HH. [Methods of anatomical and histochemical investigation of plant tissues]. M.: Nauka, 1979;40-65 p. Russian.
  8. Gnutova RV. [Serology and immunochemistry of plant viruses]. Moscow: Nauka, 1993. 300 p. Russian.
  9. Harrison SJ, Curtis MD, Mclntyre CL et al. Differential expression of peroxidase isogenes during the early stages of infection of the tropical forage legume Stylosanthes humilis by Colletotrichum gloeosporioides. Mol. Plant Microbe Interact. 1995; 8(3):398-406. https://doi.org/10.1094/MPMI-8-0398
  10. Hiraga S, Yamamoto K, Ito H. et al. Diverse expression profles of 21 rice peroxidase genes. FEBS Lett. 2000; 471:245-50. https://doi.org/10.1016/S0014-5793(00)01409-5
  11. Hrynchuk K, Antipov I, Ermolaev M. [Beet necrotic yellow vein virus: PCR-diagnostics and identifcation of Ukrainian isolates]. Quarantine and plant protection. 2015; 7(1):28-34. Ukrainian.
  12. Hrynchuk K, Antipov I, Ermolaev M. [The development of PCR system for identification of beet necrotic yellow vein virus and monitoring in Ukraine]. Scientifc Herald of Chernivtsy University. (Biological Systems). 2015; 7(1):28-34. Ukrainian.
  13. Hrynchuk K, Antipov I. [Molecular diagnostics and identifcation of the beet necrotic yellow vein virus: methodical recommendations]. Kiev. 2015; 24 p. Ukrainian.
  14. Hrynchuk K. Molecular diagnostic and identifcation of beet necrotic yellow vein virus [dissertation]. Kiev: National University of life and environmental sciences; 2016.
  15. Klein RM. Methods for studying plants. Moscow: Kolos: 1974; 527 p. 11.
  16. Klyachenko OL. Testing of beet necrotic yellow vein virus by the method of molecular hybridization of nucleic acids [dissertation]. Moscow; 1993.
  17. Lagrimini ML, Joly RJ, Dunlap JR et al. The consequence of peroxidase overexpression in transgenic plants on root growth and development. Plant Mol. Biol. 1997; 33:887-95. https://doi.org/10.1023/A:1005756713493
  18. Lucena MA, Romero-Alanda R, Mecardo JA et al. Structural and physiological changes in the roots of tomato plants over-expressing a basic peroxidase. Physiol. Plant. 2003; 118:422-29. https://doi.org/10.1034/j.1399-3054.2003.00115.x
  19. Lukyanenko T, Hrygoryuk I, Lihanov A. Peroxidase as a resistance marker of the genus Aesculus L. to horse chestnut leaf miner (Cameraria ohridella Deschka et Dimič). Quarantine and plant protection. 2014; 4:13-15.
  20. Manco OA. Rhizomania sugar beet - selection for stability. [dissertation]. Kiev: Institute of sugar beet UAAS; 2003.
  21. Moskovets SN, Bobyr AD, Glushak LE, Onishchenko AN. [Viral diseases of farm crops]. Kyiv: Urozhay, 1975; 152 p.
  22. Nurmuhammedov AK. Increasing of the stability of sugar beets to pathogens of rhizomania, rotten root crops and coronet stains: [dissertation]. Kiev: National University of life and environmental sciences; 2006.
  23. Pausheva ZP. [Workshop on cytology of plants]. Moscow: Agropromizdat. 1988; 271 p. Russian.
  24. Rasmussen J, Smith J, Williams S. et al. cDNA cloning and systemic expression of acidic peroxidases associated with systemic acquired resistance to disease in cucumber. Physiol. Mol. Plant Pathol. 1995; 46:389-400. https://doi.org/10.1006/pmpp.1995.1030
  25. Roik MV, Nurmuhammedov AK, Kornienko AS. Diseases of sugar beet root crops: coronoid stains, rotten root crops in the period of vegetation, rhizomania, non-parasitic diseases. K.: Polygraph Consulting, 2004; 213 p.
  26. Roik MV, Nurmuhammedov AK, Vasiliev NO et al. Methodical recommendations for the diagnosis of beet necrotic yellow vein virus [methodical recommendations]. Kiev: Koloobih, 2005; 27 p.
  27. Rush CM, Liu H, Lewellen RT et al. The continuing saga of Rhizomania of sugar beets in the United States. Plant Disease. 2006; 90(1):4-15. https://doi.org/10.1094/PD-90-0004
  28. Shevchenko ZhP, Helman LV, Nevyga OE, et al Virus and mycoplasma diseases of feld crops. K.: Urozhay, 1975; 152 p.
  29. Thordal-Christinsen H, Zang Z, Wei Y et al. Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley - powdery mildew interaction. Plant J. 1997; 11:1187-194. https://doi.org/10.1046/j.1365-313X.1997.11061187.x
  30. Žižytė MG. The identifcation and molecular characterization of sugar beet rhizomania causing virus: summary of doctoral dissertation, biomedical sciences, biology (01 B), microbiology, bacteriology, virology, mycology (B 230) Vilnius, 2010; 37 p.