Dipartimento di Bioscienze e Territorio, University of Molise, Pesche, IS, Italy.
Planta. 2013 Aug;238(2):271-82. doi: 10.1007/s00425-013-1890-4. Epub 2013 May 5.
Using activation tagging in Populus, we have identified five mutant lines showing changes in their adventitious rooting. Among the affected lines, three showed increased and two decreased adventitious rooting. We have positioned the tag in the mutant lines via recovering genomic sequences flanking the left-hand border of the activation tagging vector and validated the transcriptional activation of the proximal genes. We further characterized one line in which the cause of the observed rooting phenotype was up-regulation of a gene encoding a transcription factor of the AP2/ERF family of unknown function (PtaERF003). We show, through retransformation, that this gene has a positive effect on both adventitious and lateral root proliferation. Comparative expression analyses show that the phenotype does not result from ectopic expression but rather up-regulation of the native expression pattern of the gene. PtaERF003 function is linked to auxin signal transduction pathway, as suggested by the rapid induction and accentuated phenotypes of the transgenic plants in presence of the hormone. Upregulation of PtaERF003 led to most significant metabolic changes in the shoot suggesting of a broader regulatory role of the gene that is not restricted to root growth and development. Our study shows that dominant tagging approaches in poplar can successfully identify novel molecular factors controlling adventitious and lateral root formation in woody plants. Such discoveries can lead to technologies that can increase root proliferation and, thus, have significant economic and environmental benefits.
利用杨树中的激活标签技术,我们鉴定了五个表现出生根改变的突变体系。在受影响的系中,三个系显示出增加的不定根形成,两个系显示出减少的不定根形成。我们通过回收激活标签载体左边界侧翼的基因组序列,在突变体系中定位了标签,并验证了临近基因的转录激活。我们进一步对一个系进行了特征描述,该系中观察到的生根表型的原因是一个编码具有未知功能的 AP2/ERF 家族转录因子的基因上调(PtaERF003)。我们通过再转化表明,该基因对不定根和侧根增殖都有正向作用。比较表达分析表明,该表型不是由异位表达引起的,而是基因的天然表达模式的上调。PtaERF003 功能与生长素信号转导途径有关,这是由激素存在下转基因植物的快速诱导和明显表型所表明的。PtaERF003 的上调导致了芽中最显著的代谢变化,表明该基因具有更广泛的调节作用,不受根生长和发育的限制。我们的研究表明,杨树中的显性标记方法可以成功鉴定控制木本植物不定根和侧根形成的新的分子因子。这种发现可以导致增加根增殖的技术,从而具有重要的经济和环境效益。