Expression data from amiEBBB1 poplar apices

We study gene expression Populus amiEBB1 lines affecting dormancy. We used microarrays to detail the global program of gene expression underlying morphological and developmental changes droved by expression of artifical micro RNA (ami) targeting EBB1 gene. By screening activation tagging population we identified the EARLY BUD-BREAK1 (EBB1) mutant. We positioned the tag, localized a putative candidate gene and verified transcription activation. The activated gene encodes an AP2/ERF transcription factor similar to a small B1 gene-subfamily in Arabidopsis encoding strong regulators of meristem function and lateral organ outgrowth. We fully recapitulated the phenotype by overexpression of the gene into the same genotype under strong constitutive promoter (P35S::EBB1). We also found that EBB1 transcript abundance in wild type plants increases during the period prior to bud-break, and in response to the hormone cytokinin. Down-regualtion of EBB1 influence negatively bud break. Our data indicates that EBB1 plays a important role in the process of bud-break. Poplar apex was selected for RNA extraction and hybridization on Affymetrix microarrays. We used apices tissue from amiEBB1 grown greenhouse healthy plants.

本研究聚焦影响休眠的杨属amiEBB1株系的基因表达特征。我们利用基因芯片技术，解析了靶向EBB1基因的人工微RNA（amiRNA）表达所引发的形态与发育变化背后的全局基因表达调控程序。通过激活标签突变体库筛选，我们获得了EARLY BUD-BREAK1（EBB1）突变体。我们对插入标签进行了定位，鉴定出推定候选基因，并验证了其转录激活功能。该激活基因编码的AP2/ERF转录因子，与拟南芥中调控分生组织功能及侧生器官生长的小型B1基因亚家族成员具有较高同源性。我们通过将该基因在强组成型启动子P35S驱动下过表达至同一遗传背景中，成功重现了该突变表型（P35S::EBB1）。我们还发现，野生型植株中EBB1的转录本丰度在芽萌发前的阶段以及细胞分裂素处理后均显著升高。EBB1的下调会对芽萌发产生负向影响。本研究数据表明，EBB1在芽萌发过程中发挥关键作用。我们选取杨属植物茎尖进行RNA提取，并在Affymetrix基因芯片上开展杂交实验。实验所用的茎尖组织来自温室培养的健康amiEBB1株系植株。