NICOTIANA TABACUM TISSUE CULTURE AS A SOURCE OF POTATO VIRUS Y
Main Article Content
Authors
V.T. Khasanov
S. Seifullin Kazakh Agro Technical University, 62 Prospect Pobedy, Astana, 010000, Kazakhstan
Ya.I. Alekseev
Institute of Agricultural Biotechnology, 42, Timiryazeva, Moscow, 127550, Russian Federation
N.Yu. Minakova
Institute of Agricultural Biotechnology, 42, Timiryazeva, Moscow, 127550, Russian Federation
G.K. Orazbaeva
S. Seifullin Kazakh Agro Technical University, 62 Prospect Pobedy, Astana, 010000, Kazakhstan
B. Beisembina
S. Seifullin Kazakh Agro Technical University, 62 Prospect Pobedy, Astana, 010000, Kazakhstan
M.A. Fida
S. Seifullin Kazakh Agro Technical University, 62 Prospect Pobedy, Astana, 010000, Kazakhstan
Abstract
Finding alternative approaches to the technology of storage and purification of potato viruses in order to optimize the methods of their isolation is topical.
In vitro cultivation of plant callus tissue with simultaneous accumulation of a virus allows its long time accumulation, precluding other virus’s infection. Besides, the isolation of viral antigen from callus tissue provides a highly purified antigen due to absence of many specific proteins and enough amount of antigen all year round.
The purpose of our research was accumulation of potato virus Y (PVY) in plant culture tissues for further isolation and purification of viral antigen and producing specific antibodies. Accordingly, the potato virus Y was chosen as a focus of the research. Object of the research, Nicotiana tabacum Samsun variety served to accumulate the virus.
This article presents results of potato virus Y (PVY) accumulation in Nicotiana Tabacum tissue culture and its further purification. Capability of culture tissue produced from PVY-inoculated plant (Nicotiana Tabacum) to accumulate and maintain viral infection for a long time was ascertained. Purity of commercial viral preparation, as well as PVY maintaining possibility in vitro in Cherie potato plant, inoculated plants of Nicotiana tabacum and callus induced from its regenerated plants in our research were proved using ELISA and PCR in real time. In another study, mouse polyclonal antibodies specified to the virus were produced and reacted with a 1/6400 titer of antigen.
Keywords
Y-potato virus (PVY), Nicotiana tabacum, antibodies, antigen, callus, Enzyme-linked immune-sorbent assay (ELISA), PCR-method, antibody titer
Article Details
References
Agro-industrial complex of Kazakhstan. Potato production (2009) // URL (date accessed: 15.11.2012).
Guray O. How to get a good crop of potato? // URL (date accessed: 15.11.2012).
Gnutova R.V. Serologiya i immunokhimiya virusov rasteniy[Serology and immunochemistry of plant viruses]. Moscow, Nauka, 1993, 301 p.
Podderzhanie kollektsii fitopatogennykh virusov i ikh shtammov v kulture tkanei rasteniyn [Maintaining collection of pathogenic viruses and their strains in tissue culture]. Методические рекомендации ВАСХНИЛ - Guidelines VASKHNIL, 1989, vol. 7, pp. 9-11.
Clark M.F., Adams A.N. Characteristics of the mikroplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virilogy, 1977, vol. 34, pp. 475-483.
Simakov E.A., Uskov A.I. and Varitsev Y.A. Novye tekhnologii proizvodstva iskhodnogo ozdorovlennogo materiala v elitnom semenovodstve kartofelya [New technologies for production of virus-free source material in potato’s seed production], recommendation. Moscow, MSKH RF, 2000, рp. 76.
Rebrikov D.V. [PCR in Real-time. BINOM. Knowledge Laboratory]. Moscow, 2011, 223 р.
Shvidchenko V.K., Hassanov V.T., Fida M.A., Bеisеmbina B., Kharchenko P.N., Alekseev Ya. I. et al. Sravnenie metodov immunofermentnogo analiza i PCR v realnom vremeni dlya diagnostiki zarazhonnosti sortoobraztsov kartofelya virusami [Comparison of ELISA and PCR real-time for diagnosis of potato viral infection]. Journal. Vestnik Rossiyskoy akademii selskokhozyaystvennykh nauk J. Russian Academy of Agricultural Sciences, 2014, vol. 2, рр. 47-49.
Varitsev Y.A., Egorova L.I., Varitseva G.P. (1996). Shtamm Y-virusa kartofelya dlya polucheniya antigena, ispolzuemogo pri proizvodstve immunofermentnogo diagnostikuma [Strain of potato virus Y–for antigen production ELISA diagnostic]. Patent of the Russian Federation. J.Inventions №31.
Atabekov I.G., Bobkov A.F. Metodicheskie rekomendatsii po primeneniyu immunofermentnogo analiza dlya diagnostiki virusov kartofelya [Guidelines on the application of an enzyme immunoassay for the diagnosis of potato viruses]. Moscow, 1985, 19 р.
Atabekova I.G. Praktikum po obschei virusologii [Practicum on General Virology]. Moscow, publisher, MGU, 2002, 184 р.
Lebenshteyna G., Berger F.H., Brant A.A. and Lawson B.C. Virusnie i virusopodobnie bolezni i semenovodstvo kartofelya [Viral and virus-like diseases and potato’s seed production]. St. Petersburg ed. G.S., 2005, рp. 156-172.
Friemel J. Immunologicheskie metodi [Immunological Methods]. Moscow, medicine, 1987, pp. 89-97.