Detection of plants infected with citrus tristeza virus using serological methods in northern and southern regions of Iran
محورهای موضوعی : Journal of Plant ecophysiology
1 - استادیار گروه گیاهپزشکی، دانشکده کشاورزی و علوم پایه، واحد رودهن ، دانشگاه آزاد اسلامی ، رودهن، ایران
کلید واژه: Citrus tristeza virus, Diba, Western blot, serological methods,
چکیده مقاله :
Citrus tristeza virus is economically one of the most important citrus diseases in Iran. To prevent the impact of the disease in citrus growing areas, the production of virus-free plants and the removal of infected plants are key methods. In line with these goals, it is very important to obtain simple and sensitive diagnostic tools. The main goal of the present study is to develop serological methods for efficient detection of Tristeza virus in citrus cultivation areas. For this purpose, the gene encoding the coat protein (CP25 KDa) of Citrus tristeza virus was expressed in bacteria and purified through affinity chromatography. For the production of specific antibody against Tristeza virus, purified recombinant envelope protein was used to immunize rabbits, and then purification of immunoglobulin was done using protein A column. Antibodies and conjugates were made and purified, suspected plants with symptoms of citrus tristeza virus infection were collected from different parts of Mazandaran and Kerman province, ELISA, western blot and DIBA serological tests were used to identify infected plants. The results showed that the produced antibodies can be effectively used to identify plants infected with citrus tristeza virus, and out of 51 samples suspected of virus contamination, fourteen plants were definitely diagnosed with a high percentage of infection.
Citrus tristeza virus is economically one of the most important citrus diseases in Iran. To prevent the impact of the disease in citrus growing areas, the production of virus-free plants and the removal of infected plants are key methods. In line with these goals, it is very important to obtain simple and sensitive diagnostic tools. The main goal of the present study is to develop serological methods for efficient detection of Tristeza virus in citrus cultivation areas. For this purpose, the gene encoding the coat protein (CP25 KDa) of Citrus tristeza virus was expressed in bacteria and purified through affinity chromatography. For the production of specific antibody against Tristeza virus, purified recombinant envelope protein was used to immunize rabbits, and then purification of immunoglobulin was done using protein A column. Antibodies and conjugates were made and purified, suspected plants with symptoms of citrus tristeza virus infection were collected from different parts of Mazandaran and Kerman province, ELISA, western blot and DIBA serological tests were used to identify infected plants. The results showed that the produced antibodies can be effectively used to identify plants infected with citrus tristeza virus, and out of 51 samples suspected of virus contamination, fourteen plants were definitely diagnosed with a high percentage of infection.
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