School of Forest Resources and Environmental Science, Michigan Technological University, Houghton, MI 49931; and.
Department of Forest Ecosystems and Society, Oregon State University, Corvallis, OR 97331-5752.
Proc Natl Acad Sci U S A. 2014 Jul 8;111(27):10001-6. doi: 10.1073/pnas.1405621111. Epub 2014 Jun 20.
Trees from temperate latitudes transition between growth and dormancy to survive dehydration and freezing stress during winter months. We used activation tagging to isolate a dominant mutation affecting release from dormancy and identified the corresponding gene EARLY BUD-BREAK 1 (EBB1). We demonstrate through positioning of the tag, expression analysis, and retransformation experiments that EBB1 encodes a putative APETALA2/Ethylene responsive factor transcription factor. Transgenic up-regulation of the gene caused early bud-flush, whereas down-regulation delayed bud-break. Native EBB1 expression was highest in actively growing apices, undetectable during the dormancy period, but rapidly increased before bud-break. The EBB1 transcript was localized in the L1/L2 layers of the shoot meristem and leaf primordia. EBB1-overexpressing transgenic plants displayed enlarged shoot meristems, open and poorly differentiated buds, and a higher rate of cell division in the apex. Transcriptome analyses of the EBB1 transgenics identified 971 differentially expressed genes whose expression correlated with the EBB1 expression changes in the transgenic plants. Promoter analysis among the differentially expressed genes for the presence of a canonical EBB1-binding site identified 65 putative target genes, indicative of a broad regulatory context of EBB1 function. Our results suggest that EBB1 has a major and integrative role in reactivation of meristem activity after winter dormancy.
温带地区的树木在冬季会经历生长和休眠的交替,以适应脱水和冻结的压力。我们使用激活标签来分离影响休眠解除的显性突变,并鉴定了相应的基因 EARLY BUD-BREAK 1(EBB1)。通过标记定位、表达分析和转基因恢复实验,我们证明 EBB1 编码一个假定的 APETALA2/乙烯响应因子转录因子。该基因的过表达导致芽的早期萌发,而过表达则延迟芽的萌发。EBB1 的表达在活跃生长的顶端最高,休眠期无法检测到,但在芽突破前迅速增加。EBB1 转录本定位于茎尖分生组织和叶原基的 L1/L2 层。EBB1 过表达的转基因植物表现出增大的茎尖分生组织、开放且分化不良的芽,以及更高的顶端细胞分裂率。EBB1 转基因植物的转录组分析鉴定了 971 个差异表达基因,其表达与转基因植物中 EBB1 表达变化相关。对差异表达基因的启动子分析发现了 65 个可能的 EBB1 结合位点,表明 EBB1 功能具有广泛的调节背景。我们的结果表明,EBB1 在冬季休眠后重新激活分生组织活性方面具有主要和综合的作用。