PERTANIKA JOURNAL OF TROPICAL AGRICULTURAL SCIENCE

 

e-ISSN 2231-8542
ISSN 1511-3701

Home / Regular Issue / JTAS Vol. 39 (2) May. 2016 / JTAS-0662-2014

 

Accumulation of Potato Virus Y- in Nicotiana tabacum Callus Culture to Obtain a Virus Preparation

Khassanov, V. T., Beisembina, B. and Fida, M. A.

Pertanika Journal of Tropical Agricultural Science, Volume 39, Issue 2, May 2016

Keywords: Potato virus Y- (PVY), callus tissue, ELISA, antigen, polyclonal antibodies

Published on: 11 Apr 2016

The accumulation of potato virus Y- (PVY) in tissue culture of Nicotiana tabacum (N. tabacum) was studied. Plants of N. tabacum variety, Samsun, inoculated with PVY-infected sap of potato variety, Cherie, were used for the virus accumulation. According to enzyme-linked immunosorbent assay (ELISA) results, virus showed the highest optical density (OD) on the 25th day of inoculation. Murashige and Skoog medium containing kinetin 2 mg/l, 2.4-D 0.5 mg/l, indole-acetic acid 1 mg/l sucrose 2% agar 0.7%, in addition to the standard components, was used to induce callus culture from N. tabacum leave explants. ELISA results showed that the callus culture was able to maintain viral infection during four transplantations. Slightly and highly purified (Y-Cherie) virus preparations were obtained from the PVY-infected tissue culture. The slightly-purified antigens showed an OD approximately equal to the positive control in sandwich ELISA. The Y-Cherie antigen was detected as PVY necrotic strain. Specific to the virus polyclonal antibodies that reacted with a maximum 1/3200 titer of antigen in indirect ELISA were obtained in the result of the laboratory mouse immunisation.

ISSN 1511-3701

e-ISSN 2231-8542

Article ID

JTAS-0662-2014

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