PRODUCTION OF POLYCLONAL ANTISERUM AGAINST THE CAUSAL AGENT OF LIME WITCHES'-BROOM

MIRZAI, M.; HEYDARNEJAD, J.; SALEHI, M.; HOSSEINI POUR, A.; MASSUMI, H.; SHAABANIAN, M.

PRODUCTION OF POLYCLONAL ANTISERUM AGAINST THE CAUSAL AGENT OF LIME WITCHES'-BROOM

Document Type : Research Article

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Abstract

Lime witches’-broom (LWB) caused by Candidatus Phytoplasma aurantifolia is one of the most destructive diseases of lime in the southern Provinces of Iran. To prepare LWB polyclonal antibody, midribs of the infected plants were homogenized using GMS buffer. After low and high speed centrifugations, the extract was incubated with an antiserum previously prepared against healthy lime extract. Differential centrifugation was repeated and the final extract was emulsified in equal volume of freund’s incomplete adjuvant and injected into New Zealand white rabbits. After purification of gamma globulin, the best reaction was found between the 1/100 dilution of the gamma globulin and the 1/500 dilution of infected plant extract in indirect ELISA. Furthermore, the 1/100 dilution of the gamma globulin was reacted with the 1/100 and 1/500 dilutions of infected plant extract in DIBA test. Whereas no reaction was found between the antiserum and healthy lime extracts. The results indicated that antibody raised against Candidatus Phytoplasma aurantifolia could be used for monitoring LWB disease in the field.

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