Mikrobiol. Z. 2018; 80(1):77-88. Ukrainian.
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.
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