S.G. Vologin, I.V. Pikalova, Z. Stashevski, M.Sh. Tagirov
Tatar Agriculture Research Institute, Orenburgsky Trakt Str., 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 find the optimal method for biomaterial preparation, which included the application of buffer and antiseptic compounds and increased the accuracy of potato virus Y (PVY) detection by enzyme-linked immunoassay (EIA). Application of 0.1 M Tris-HCl buffer (TB) was sufficient to maintain the acid-base balance in the potato material. Introduction of 0.01 M sodium azide together with 0.1 M TB into biomaterial led to the decrease in the level of detectable colorimetric signal during the EIA in the test of leaf samples by 59.6 % (p < 0.01), and of etiolated tuber shoots - by 7.6 % (p > 0.05). It was supposed that pre-analytical effect of sodium azide was associated with oxidative damage of PVY coat protein. During examination of leaf and tuber samples the application of 0.01 sodium merthiolate (SM) in combination with 0.1 M TB resulted in a statistically significant increase in the level of detectable signal, respectively, by 38.3 and 62.9 % (p < 0.01). It was suggested that the detected effect associated with the biochemical transformation of SM in plant material leading to the formation of thiosalicylic acid (TSA). This compound includes a thiogroup, which may block the negative effect of a number of polyphenolic compounds. Adding of 0.01 M TSA at the stage of sample preparation was accompanied by an increase in the level of the detected signal (p < 0.01). The positive effect of TSA was comparable with the effect of other thiogroup-containing compounds - thioglycolic acid and 2-mercaptoethanol, as well as of substances with antioxidant (sodium sulfite, ascorbic acid), chelating (EDTA) and phenol binding (polyvinyl pyrrolidone) properties. Application of SM and TSA provided reliable PVY immunodiagnostic during 7 days of preanalytical storage at + 2 ... + 8 deg. To increase the significance of potato virus diagnostic results it was recommended to use 0.1 M TB in combination with 0.01 M SM or 0.01 M TSA at the stage of the preparation of biomaterial.
Keywords: diagnosis, potato, sample preparation, potato virus, enzyme-linked immunoassay, sodium azide, sodium merthiolate, thiosalicylic acid.
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 stabilizing compounds. Dostizheniya nauki i tekhniki APK. 2015. V.29. №5. pp. 47-50 (In Russ)