Evaluation of certain physicochemical and biological characteristics of Beet necrotic yellow vein virus (BNYVV) and preparation of antiserum against it
2005
Darabi, S. | Bazrafshan, M. | Khajeh, A. | Izad-Panah, K.
Rhizomania is a very damaging disease of sugar beet worldwide. In this research, in order to use of serological methods to detect of disease as well as for screening tolerant plant materials, BNYVV was purified and antiserum was prepared against it. The virus was propagated on Chenopodium quinoa leaves by mechanical inoculation of infected sugar beet leaves. Purification of BNYVV from C.quinoa involved extraction of infected tissue in phosphate buffer, primary clarification, centrifugation at high speed through a cushon of 20% sucrose and centrifugation at high speed through a sucrose density gradient. Then, light-scattering band of virus particles was detected in centrifuge tube. For preparation of antiserum, purified virus was injected subcutaneously into a NewZeland rabbit. DAS-ELISA system was developed after isolation of ã-globulin from absorbed antiserum and preparation of enzyme-antibody conjugate. Agarose gel electrophoresis of BNYVV purified RNA revealed three band at 2.22 106 Da RNA1 position, 1.65 106 Da RNA2 position and 0.81 106 Da RNA3 position.
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