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Transgenic resistance to <Emphasis Type="Italic">Cymbidium mosaic virus</Emphasis> in <Emphasis Type="Italic">Dendrobium</Emphasis> expressing the viral capsid protein gene
Authors:Chen?Chang  Ying-Chun?Chen  Yau-Heiu?Hsu  Jian-Tai?Wu  Chung-Chi?Hu  Wei-Chin?Chang  Email author" target="_blank">Na-Sheng?LinEmail author
Institution:(1) Department of Botany, National Museum of Natural Science, 404, Taichung, Taiwan, ROC;(2) Institute of Botany, Academia Sinica, Taiwan, Taipei, ROC, 115;(3) Graduate Institute of Agricultural Biotechnology, National Chung Hsing University, 402, Taichung, Taiwan, ROC
Abstract:A Taiwan isolate of Cymbidium mosaic virus (CymMV-CS) was isolated from infected Cymbidium sinesis Willd. The cDNA of the capsid protein (CP) gene was synthesized and sequenced. Alignment of this CP gene with other reported CPs revealed homologies of 92–98% at the nucleotide level and 98–99% at the amino acid level. To generate virus-resistant varieties, the CymMV-CS CP gene was transformed into Dendrobium protocorms through particle bombardment. Transformants were selected on medium supplemented with 20 mg/L hygromycin and the presence of the transgene was confirmed by polymerase chain reaction, Southern, Northern and Western blot analyses. Transgenic Dendrobium harboring the CymMV CP gene expressed a very low level of virus accumulation four months post-inoculation with CymMV, as detected by ELISA. The transgenic plants exhibited much milder symptoms than the non-transgenic plants upon challenge with CymMV virionsSequence data reported from this article have been deposited at the GenBank Data Libraries under Accession No. AY429021.
Keywords:capsid protein (CP)  Cymbidium mosaic virus (CymMV)  particle bombardment  transgenic Dendrobium  virus resistance
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