Virus-induced gene silencing in tomato

被引:1280
作者
Liu, YL [1 ]
Schiff, M [1 ]
Dinesh-Kumar, SP [1 ]
机构
[1] Yale Univ, Dept Mol Cellular & Dev Biol, OML 451, New Haven, CT 06520 USA
关键词
tobacco rattle virus vector; gene silencing; tomato; tCTR1; functional genomics; GATEWAY cloning;
D O I
10.1046/j.1365-313X.2002.01394.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
We have previously demonstrated that a tobacco rattle virus (TRV)-based vector can be used in virus-induced gene silencing (VIGS) to study gene function in Nicotiana benthamiana . Here we show that recombinant TRV infects tomato plants and induces efficient gene silencing. Using this system, we suppressed the PDS , CTR1 and CTR2 genes in tomato. Suppression of CTR1 led to a constitutive ethylene response phenotype and up-regulation of an ethylene response gene, CHITINASE B . This phenotype is similar to Arabidopsis ctr1 mutant plants. We have constructed a modified TRV vector based on the GATEWAY recombination system, allowing restriction- and ligation-free cloning. Our results show that tomato expressed sequence tags (ESTs) can easily be cloned into this modified vector using a single set of primers. Using this vector, we have silenced RbcS and an endogenous gene homologous to the tomato EST cLED3L14. In the future, this modified vector system will facilitate large-scale functional analysis of tomato ESTs.
引用
收藏
页码:777 / 786
页数:10
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