Production of siRNA targeted against TYLCV coat protein transcripts leads to silencing of its expression and resistance to the virus.
The coat protein (CP) of Tomato yellow leaf curl virus (TYLCV), encoded by the v1 gene, is the only known component of the viral capsid. In addition, the CP plays a role in the virus transport into the host cell nucleus where viral genes are replicated and transcribed. In this study, we analyzed the effect of small interfering double-stranded RNAs (siRNAs), derived from an intron-hairpin RNA (ihpRNA) construct and targeting the v1 gene product, on CP accumulation. Transient assays involving agroinfiltration of the CP-silencing construct followed by infiltration of a fused GFP-CP (green fluorescent protein-coat protein) gene showed down-regulation of GFP expression in Nicotiana benthamiana. Some of the transgenic tomato plants (cv. Micro-Tom), expressing the siRNA targeted against the TYLCV CP gene, did not show disease symptoms 7 weeks post-inoculation with the virus, while non-transgenic control plants were infected within 2 weeks post inoculation. The present study demonstrates, for the first time, that siRNA targeted against the CP of TYLCV can confer resistance to the virus in transgenic tomato plants, thereby enabling flowering and fruit production.
Zrachya
Avi
A
Department of Genetics, Agricultural Research Organization, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel.
Kumar
Pravin P
PP
Ramakrishnan
Usha
U
Levy
Yael
Y
Loyter
Abraham
A
Arazi
Tzahi
T
Lapidot
Moshe
M
Gafni
Yedidya
Y
D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000502:physiology; D036022:Capsid Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D018551:Solanum lycopersicum / Q000166:cytology / Q000235:genetics / Q000821:virology*; D018613:Microscopy, Confocal; D010935:Plant Diseases / Q000235:genetics* / Q000821:virology*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000821:virology; D020871:RNA Stability; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics; D014158:Transcription, Genetic
D036022:Capsid Proteins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins