Duval Isabelle, Lachance Denis, Giguère Isabelle, Bomal Claude, Morency Marie-Josée, Pelletier Gervais, Boyle Brian, MacKay John J, Séguin Armand
Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, Québec, QC, G1V 4C7, Canada.
Centre d'Étude de la Forêt, Université Laval, Québec, QC, G1V A06, Canada.
J Exp Bot. 2014 Jun;65(9):2319-33. doi: 10.1093/jxb/eru116. Epub 2014 Apr 8.
This research aimed to investigate the role of diverse transcription factors (TFs) and to delineate gene regulatory networks directly in conifers at a relatively high-throughput level. The approach integrated sequence analyses, transcript profiling, and development of a conifer-specific activation assay. Transcript accumulation profiles of 102 TFs and potential target genes were clustered to identify groups of coordinately expressed genes. Several different patterns of transcript accumulation were observed by profiling in nine different organs and tissues: 27 genes were preferential to secondary xylem both in stems and roots, and other genes were preferential to phelloderm and periderm or were more ubiquitous. A robust system has been established as a screening approach to define which TFs have the ability to regulate a given promoter in planta. Trans-activation or repression effects were observed in 30% of TF-candidate gene promoter combinations. As a proof of concept, phylogenetic analysis and expression and trans-activation data were used to demonstrate that two spruce NAC-domain proteins most likely play key roles in secondary vascular growth as observed in other plant species. This study tested many TFs from diverse families in a conifer tree species, which broadens the knowledge of promoter-TF interactions in wood development and enables comparisons of gene regulatory networks found in angiosperms and gymnosperms.
本研究旨在探讨多种转录因子(TFs)的作用,并在相对高通量的水平上直接描绘针叶树中的基因调控网络。该方法整合了序列分析、转录谱分析以及针叶树特异性激活分析的开发。对102个转录因子和潜在靶基因的转录积累谱进行聚类,以识别协同表达的基因群体。通过在九个不同器官和组织中的分析,观察到几种不同的转录积累模式:27个基因在茎和根中都优先表达于次生木质部,其他基因则优先表达于栓内层和周皮,或者分布更为广泛。已经建立了一个强大的系统作为筛选方法,以确定哪些转录因子能够在植物中调控给定的启动子。在30%的转录因子-候选基因启动子组合中观察到了反式激活或抑制作用。作为概念验证,系统发育分析以及表达和反式激活数据被用于证明,正如在其他植物物种中所观察到的那样,两种云杉NAC结构域蛋白很可能在次生维管生长中起关键作用。本研究在一种针叶树种中测试了来自不同家族的许多转录因子,这拓宽了对木材发育中启动子-转录因子相互作用的认识,并能够比较被子植物和裸子植物中的基因调控网络。