Transgenic resistance to Cymbidium mosaic virus in Dendrobium expressing the viral capsid protein gene.
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 virions.
Chang
Chen
C
Department of Botany, National Museum of Natural Science, Taichung, Taiwan 404, ROC.
Chen
Ying-Chun
YC
Hsu
Yau-Heiu
YH
Wu
Jian-Tai
JT
Hu
Chung-Chi
CC
Chang
Wei-Chin
WC
Lin
Na-Sheng
NS
D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics*; D017931:DNA Primers; D031667:Dendrobium / Q000235:genetics / Q000821:virology*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000821:virology*
D036022:Capsid Proteins; D017931:DNA Primers