Purification and antiserum production of Iranian isolate of Beet Necrotic Yellow Vein Virus (BNYVV), the causal agent of sugar beet rhizomania disease
2010
S. Darabi (Instructor of Sugar Beet Research Department, Fars Agricultural and Natural Resources Research Center, AREEO, Shiraz, Iran.) | M. Yassaie (Instructor of Agricultural and Natural Resources Research Centre- Seed and Plant Improvement Department, Fars, Iran) | K. Izadpanah (Professor of Plant Virology Research Center, College of Agriculture, Shiraz University, Shiraz, Iran.)
Beet Necrotic Yellow Vein Virus (BNYVV) is the causal agent of sugar beet rhizomania, disease . In the present study which was conducted in 2005 in Shiraz, Iran, serological methods (including ELISA) were used for detection of disease and screening of disease tolerant germplasm in sugar beet breeding programs. For doing this, Iranian isolate of BNYVV was purified and a specific antiserum was prepared against it. The virus was propagated on Chenopodium quinoa leaves by mechanical inoculation of sap extracted from the infected sugar beet leaves. Purification of BNYVV from C. quinoa comprised of extraction of infected sap in phosphate buffer, primary clarification, high speed centrifugation through a cushion of 20% sucrose and sucrose density gradient centrifugation. The light-scattering band of virus particles was formed in centrifuge tubes with more densitymainly in three sections. For preparation of antiserum, the purified virus was injected subcutaneously into a New Zeland rabbit. DAS-ELISA system was developed after isolation of γ-globulin from absorbed antiserum and preparation of enzyme-antibody conjugate. This antiserum could be used in the related research at education and research organizations. The antiserum produced was of high quality and did not react with the healthy plant sap.
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