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Citrus psorosis is probably caused by a bipartate ssRNA virus
Institution:1. Central Region Agricultural Research Center, NARO, Kannondai, Tsukuba, Ibaraki 305-8666, Japan;2. Kochi Agricultural Research Center, Hataeda, Nankoku, Kochi 783-0023, Japan;3. Aichi Agricultural Research Center, Sagamine, Yazako, Nagakute, Aichi 480-1193, Japan;1. Department of International Agricultural Technology and Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Republic of Korea;2. Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Republic of Korea;1. Chair of Plant Physiology and Biotechnology, Nicolaus Copernicus University, Lwowska 1, 87-100 Toruń, Poland;3. Department of Microbiology, Nicolaus Copernicus University, Lwowska 1, 87-100 Toruń, Poland;1. Lehrstuhl für Technische Mikrobiologie, Technische Universität München (TUM), Freising, Germany;2. Institute of Applied Biosciences, Department of Food Chemistry and Phytochemistry, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany;1. Spanish National Centre for Biotechnology (CSIC), Madrid, Spain;2. Grupo Interdisciplinar de Sistemas Complejos (GISC), Madrid, Spain
Abstract:Isolate 90-1-1 Concordia (Argentina) of the citrus psorosis agent was graft-transmitted to citrus and mechanically transmitted to Chenopodium quinoa, which was used as a local lesion assay host. Infected citrus and C. quinoa plant lesions were used as starting materials for the purification of the psorosis-associated agent.In extracts partially purified by differential centrifugation, infectivity was abolished by RNase treatment, even in 0.3 M NaCl, indicating that ssRNA is required for biological activity. The total loss of infectivity produced by proteinase K treatment and the decline in infectivity caused by phenol extraction indicated that protein may be essential for infectivity.When partially purified extracts were subjected to sucrose density gradient centrifugation, infectivity on C. quinoa from certain 2-fraction combinations was higher than expected, compared to the infectivity of the individual fractions. Therefore, infectivity was not associated with a single component but with the combination of at least two components which were distinguishable on sedimentation.The infectious material was present in the top and bottom zones of a sucrose gradient, which, on further purification by a second gradient and agarose gel electrophoresis, revealed the presence of a 50-kDa protein. This protein was absent in comparable gradient fractions from healthy plants, and therefore most likely represented the capsid protein of both the top and bottom sucrose gradient zone components.Taken together, these results led to the conclusion that the citrus-psorosis-associated virus (CPsAV) is a multipartite virus, containing ssRNA and a 50-kDa coat protein. In view of the information available to date, CPsAV seems to be very closely related to citrus ringspot virus described in Florida (USA) and the psoriasis agent in Spain.
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