S.G. Vologin, I.V. Pikalova, Z. Stashevski, M.Sh. Tagirov
Tatar Agriculture Research Institute, Orenburgsky trakt, 48, Kazan, 420059, Russia
E-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.
Summary. The study was carried out to assess complex effect of potato biological material and storage temperature on results of diagnostic of Potato virus Y (PVY) by enzyme-linked immunoassay (EIA). At examination of fractional composition of bioassay, prepared for the diagnostic, it was found that the mass fraction of destroyed plant components (DPC) not merged in fraction used in the diagnostics in leaf samples was in the range of 15…20 %, and in tuber sprout samples – at 45…50 %. The main elements of DPC in the leaf samples were chloroplasts, and in etiolated sprouts it was starch grains. Introduction of the biomaterial, containing DPC and stored at +2 ... + 8 degrees, to the diagnostic analysis resulted in decrease in the level of detectable colorimetric signal in EIA in the examination of leaf samples on average by 24.8% and tuber samples – on 47.7%. This suggested inhibition of the reaction "antigen-antibody". Similar inhibition was also observed when using etiolated tuber shoots containing DCP and once subjected to the procedure of freezing-thawing (-18 ... -25 deg). At the same time, the application of leaf samples subjected to freezing-thawing cycle, resulted in the increase in detectable signal level on the average by 18.8 %. In the course of the experiments it was registered the output of additional amount of the virus from the DPC of leaf samples during low-temperature processing. To improve the accuracy of the diagnostic result of viral pathogens in potato leaf samples it is recommended to carry out freezing-thawing cycle of destroyed tissues and their subsequent removal by centrifugation immediately prior to EIA. With this method of sample preparation it was noted twofold increase in detectable colorimetric signal level in the EIA.
Keywords: diagnosis, potato, sample preparation, potato viruses, enzyme-linked immunoassay.
Author Details: S.G. Vologin, Сand. Sc. (Biol.), Head of Laboratory (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); I.V. Pikalova, Researcher; Z. Stashevski, Сand. Sc. (Biol.), Head of Department; M.Sh. Tagirov, Dr. Sc. (Agr.), Director.
For citation: Vologin S.G., Pikalova I.V., Stashevski Z., Tagirov M.Sh. Biomaterial preparation for potato virus diagnostic: effect of samplе composition and storage temperature. Dostizheniya nauki i tekhniki APK. 2015. V.29. №5. pp. 43-46 (In Russ)