PRODUCTION AND EVALUATION OF SPECIFIC ANTIBODIES RAISED AGAINST A NEW ISOLATE OF TOMATO YELLOW LEAF CURL GEMINIVIRUS INFECTING SUGAR BEET
Abstract
Sugar beet is considered as the second important crop for sugar production worldwide. The major problem associated with sugar beet production, its high vulnerability to at least sixteen different viruses. Therefore, production of efficient and specific diagnostic tools is highly required for rapid and convenient virus detection. This study aimed to better characterization of a new isolate of the tomato yellow leaf curl geminivirus infecting sugar beet plants in Egypt, followed by producing of specific diagnostic antibodies to facilitate the viral detection in infected plants. The presence of the virus isolate in infected beet plants were tested using whitefly transmitting geminivirus specific primers. Viruleferous whiteflies were used to infect healthy beet plants to obtain sufficient amount required for analysis. Virus monitoring in beet plants was characterized by PCR using whitefly transmitting geminivirus (WTG) specific primers which confirmed the viral presence in DNA extracted from infected beet plants. Further molecular characterization was performed by sequencing the PCR product and it was blasted to the GenBank for generating phylogenetic tree, which proved that the obtained virus sequence is close to the TYLCV isolates. This suggested that it could be new isolate related to the published TYLCV isolates. Furthermore, Serological studies were performed using total proteins extracted from the infected Beet plants after symptoms manifestation including curling and upward cup shape of the leaves in 40-45 days post infection. Virus capsid protein band was extracted and used for mice injection to raise viral specific antibodies. Efficacy of the obtained viral antiserum for virus infection diagnoses was tested and confirmed the use of ELISA and western blotting for this purpose.
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