TY - JOUR
T1 - Biologically active cymbidium ringspot virus satellite RNA in transgenic plants suppresses accumulation of DI RNA
AU - Rubino, Luisa
AU - Carrington, James C.
AU - Russo, Marcello
N1 - Funding Information:
L.R. and M.R. thank Deon D. Freed for introducing them to plant transformation techniques, Antonella Antonacci for the skillful technical help given during the course of this work, and Professor G. P. Martelli for helpful discussion. This research was supported by National Research Council of Italy, Special Project RAISA, Subproject No. 2, Paper No. 295, and by a grant to J.C.C. from the National Institutes of Health (Al27832).
PY - 1992/6
Y1 - 1992/6
N2 - A full-length DNA copy of cymbidium ringspot virus (CyRSV) satellite RNA was cloned downstream of the bacteriophage T7 RNA polymerase promoter. In vitro transcripts were biologically active in plants when coinoculated with the helper virus or its RNA. Although the transcripts contained 7 or 29 extra nucleotides at the 3′ end, the proper 3′ terminus was restored in the satRNA progeny. Full-length cDNA clones of CyRSV satRNA under the control of the cauliflower mosaic virus 35S promoter and terminator were used to transform Nicotiana benthamiana plants. Integration of CyRSV satRNA sequence in the plant genome was tested by PCR amplification of DNA extracts from transformed plants and by detection of satRNA-related transcripts in total RNA extracts. Inoculation of transgenic plants with the helper virus induced replication of satRNA of the same size as the native molecule. Sequence analysis of the satRNA progeny showed that it was identical to natural CyRSV satRNA. Infected transgenic plants were not protected from apical necrosis and death by the presence of satRNA sequences. Rather, replication of satRNA was found to suppress accumulation of defective interfering RNA, which acts in the absence of satRNA as an attenuator of virus replication and disease.
AB - A full-length DNA copy of cymbidium ringspot virus (CyRSV) satellite RNA was cloned downstream of the bacteriophage T7 RNA polymerase promoter. In vitro transcripts were biologically active in plants when coinoculated with the helper virus or its RNA. Although the transcripts contained 7 or 29 extra nucleotides at the 3′ end, the proper 3′ terminus was restored in the satRNA progeny. Full-length cDNA clones of CyRSV satRNA under the control of the cauliflower mosaic virus 35S promoter and terminator were used to transform Nicotiana benthamiana plants. Integration of CyRSV satRNA sequence in the plant genome was tested by PCR amplification of DNA extracts from transformed plants and by detection of satRNA-related transcripts in total RNA extracts. Inoculation of transgenic plants with the helper virus induced replication of satRNA of the same size as the native molecule. Sequence analysis of the satRNA progeny showed that it was identical to natural CyRSV satRNA. Infected transgenic plants were not protected from apical necrosis and death by the presence of satRNA sequences. Rather, replication of satRNA was found to suppress accumulation of defective interfering RNA, which acts in the absence of satRNA as an attenuator of virus replication and disease.
UR - http://www.scopus.com/inward/record.url?scp=0026772591&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(92)90496-C
DO - 10.1016/0042-6822(92)90496-C
M3 - Article
C2 - 1374981
AN - SCOPUS:0026772591
SN - 0042-6822
VL - 188
SP - 429
EP - 437
JO - Virology
JF - Virology
IS - 2
ER -