TY - JOUR
T1 - Analysis of the nucleocapsid protein gene from Tomato spotted wilt virus as target and inducer for posttranscriptional gene silencing
AU - Sonoda, Shoji
N1 - Funding Information:
This study was supported in part by the Grant-in-Aid for Encouragement of Young Scientists from the Ministry of Education, Culture, Sports, Science and Technology (No. 13760037) and the Ohara Foundation for Agricultural Research. I thank Dr D.C. Baulcombe (The Sainsbury Laboratory, Norwich, UK) for providing the PVX vector; Dr S. Tsuda (National Agricultural Research Center, Tsukuba, Japan) for providing Tomato spotted wilt virus; Dr M. Nishiguchi (Ehime University, Matsuyama, Japan) for reading the manuscript.
PY - 2003/5/1
Y1 - 2003/5/1
N2 - Nicotiana benthamiana was transformed with the nucleocapsid protein (N) gene of Tomato spotted wilt virus (TSWV). Out of 17 transgenic lines regenerated, 6 showed virus resistance against TSWV and recombinant Potato virus X containing the N gene (PVX.N) and accumulated relatively low steady-state levels of mRNA of the N transgene, suggesting that the resistance was manifested by a posttranscriptional gene silencing. The targets for RNA degradation in the resistant lines were mainly concentrated in the 3′ portion. Non-transgenic N. benthamiana infected with PVX.N showed resistance to TSWV. On the other hand, no resistance to TSWV was observed in plants infected with PVX containing a sequence non-homologous to TSWV. These results suggest that the protection was RNA-mediated. Both 5′ and 3′ portions of the N gene were shown to be efficient inducers for the RNA-mediated cross-protection. When a non-silenced line was inoculated with recombinant PVX containing various regions of the N gene, virus-induced gene silencing (VIGS) of the N transgene mRNA was observed. Furthermore, it was shown that the 5′ portion of the N gene induced VIGS more efficiently than the 3′ portion.
AB - Nicotiana benthamiana was transformed with the nucleocapsid protein (N) gene of Tomato spotted wilt virus (TSWV). Out of 17 transgenic lines regenerated, 6 showed virus resistance against TSWV and recombinant Potato virus X containing the N gene (PVX.N) and accumulated relatively low steady-state levels of mRNA of the N transgene, suggesting that the resistance was manifested by a posttranscriptional gene silencing. The targets for RNA degradation in the resistant lines were mainly concentrated in the 3′ portion. Non-transgenic N. benthamiana infected with PVX.N showed resistance to TSWV. On the other hand, no resistance to TSWV was observed in plants infected with PVX containing a sequence non-homologous to TSWV. These results suggest that the protection was RNA-mediated. Both 5′ and 3′ portions of the N gene were shown to be efficient inducers for the RNA-mediated cross-protection. When a non-silenced line was inoculated with recombinant PVX containing various regions of the N gene, virus-induced gene silencing (VIGS) of the N transgene mRNA was observed. Furthermore, it was shown that the 5′ portion of the N gene induced VIGS more efficiently than the 3′ portion.
KW - Cross-protection
KW - Gene silencing
KW - Nicotiana benthamiana
KW - Tomato spotted wilt virus
KW - Virus resistance
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U2 - 10.1016/S0168-9452(03)00028-1
DO - 10.1016/S0168-9452(03)00028-1
M3 - Article
AN - SCOPUS:0038068138
SN - 0168-9452
VL - 164
SP - 717
EP - 725
JO - Plant Science
JF - Plant Science
IS - 5
ER -