Virus-induced gene silencing in tomato
Virus-induced gene silencing in tomato
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DOI：
10.1046/j.1365-313x.2002.01394.x
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发表时间：
2002-09-01
期刊：
影响因子：
7.2
通讯作者：
Dinesh-Kumar, SP
中科院分区：
文献类型：
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作者：
Liu, YL;Schiff, M;Dinesh-Kumar, SP
我们先前已经证明，基于烟草摇铃病毒(TRV)的载体可以用于病毒诱导的基因沉默(VIGS)来研究本氏烟草的基因功能。在这里，我们证明了重组TRV侵染番茄植株并诱导有效的基因沉默。利用该系统，我们对番茄中的PDS、CTR1和CTR2基因进行了抑制。抑制CTR1导致了结构性的乙烯反应表型和乙烯反应基因几丁质酶B的上调。该表型与拟南芥CTR1突变株相似。我们构建了一个基于网关重组系统的改进的TRV载体，允许无限制和连接的克隆。我们的结果表明，只需一组引物，番茄表达序列标签(EST)就可以很容易地克隆到这个修改的载体中。利用这个载体，我们已经沉默了RBC和一个与番茄EST cLED3L14同源的内源基因。这一改良的载体系统将为番茄EST的大规模功能分析奠定基础。
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.