Virus induced gene silencing of a DEFICIENS ortholog in Nicotiana benthamiana
Virus induced gene silencing of a DEFICIENS ortholog in Nicotiana benthamiana
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DOI：
10.1023/b:plan.0000040899.53378.83
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发表时间：
2004-03-01
影响因子：
5.1
通讯作者：
Dinesh-Kumar, SP
中科院分区：
文献类型：
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作者：
Liu, Y;Nakayama, N;Dinesh-Kumar, SP
传统上，植物生物学的发育研究因缺乏研究非模式植物物种基因功能的便捷方法而受阻。在此我们表明，病毒诱导的基因沉默（VIGS）是一种有效的新工具，可用于研究非模式系统中花同源异型基因（如DEFICIENS（DEF））同源基因的功能。我们使用基于烟草脆裂病毒（TRV）的VIGS方法来研究本氏烟草DEF同源基因（NbDEF）的功能。利用TRV - VIGS使本氏烟草中NbDEF沉默的情况与金鱼草def和拟南芥ap3突变体相似，并导致花瓣转变为萼片，雄蕊转变为心皮。对NbDEF沉默植株的分子分析显示，花朵中NbDEF的mRNA和蛋白质水平显著降低。NbDEF沉默是特异性的，对NbDEF最接近的旁系同源基因NbTM6的mRNA水平没有影响。在NbDEF沉默植株的花朵中还观察到本氏烟草GLOBOSA（NbGLO）的mRNA和蛋白质水平显著降低，这表明NbDEF对这个类似GLO的基因存在交叉调控。综上所述，我们的结果表明NbDEF是金鱼草DEF的功能同源物。我们的研究结果意义重大，因为它们表明TRV能有效地诱导幼嫩和正在分化的花朵中的基因沉默，并且VIGS是分析非模式生物中发育基因功能的一种有前途的新工具。
Traditionally, developmental studies in plant biology have suffered from the lack of a convenient means to study gene function in non-model plant species. Here we show that virus-induced gene silencing (VIGS) is an effective new tool to study the function of orthologs of floral homeotic genes such as DEFICIENS (DEF) in non-model systems. We used a tobacco rattle virus (TRV)-based VIGS approach to study the function of the Nicotiana benthamiana DEF ortholog (NbDEF). Silencing of NbDEF in N. benthamiana using TRV-VIGS was similar to that of Antirrhinum def and Arabidopsis ap3 mutants and caused transformation of petals into sepals and stamens into carpels. Molecular analysis of the NbDEF-silenced plants revealed a dramatic reduction of the levels of NbDEF mRNA and protein in flowers. NbDEF silencing was specific and has no effect on the mRNA levels of NbTM6, the closest paralog of NbDEF A dramatic reduction of the levels of N. benthamiana GLOBOSA (NbGLO) mRNA and protein was also observed in flowers of NbDEF-silenced plants, suggesting that cross-regulation of this GLO-like gene by NbDEF Taken together, our results suggest that NbDEF is a functional homolog of Antirrhinum DEF. Our results are significant in that they show that TRV efficiently induces gene silencing in young and differentiating flowers and that VIGS is a promising new tool for analyses of developmental gene function in non-model organisms.