逆境下ERF转录因子调控基因表达所结合的关键顺式元件及抗逆调控机理研究

项目名称： 逆境下ERF转录因子调控基因表达所结合的关键顺式元件及抗逆调控机理研究

项目编号： No.31470663

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 王玉成

作者单位： 东北林业大学

项目金额： 90万元

中文摘要： 我们对柽柳的一条ERF基因ThERF1研究发现，仅20%的ThERF1调控基因启动子中有GCC-box或DRE元件，对柽柳其他ERF研究也有类似现象。进一步研究显示逆境下ERF可能结合其他元件调控基因表达，GCC-box和DRE不起主要作用。有鉴于此，本研究拟将抗逆ERF转入柽柳，对转基因柽柳进行正常、盐、旱、ABA处理，分别建立ChIP文库，进行ChIP-Seq高通量测序，获得ERF在不同逆境及非逆境下结合的DNA片段序列，进行序列的保守元件分析，鉴定出胁迫及非胁迫下ERF可能结合的元件，酵母单杂交验证。分析这些元件在胁迫和非胁迫下出现频率，鉴定出逆境下ERF在表达调控中主要结合的元件。建立逆境和非逆境下柽柳转录组，通过转录组与ChIP序列的深度拼接，阐明这些元件可能存在于哪些基因启动子中，参与了何种抗逆代谢途径的调控，进而鉴定出逆境胁迫下ERF所调控的关键元件及其所参与的抗逆调控机理。

中文关键词： 柽柳；ERF转录因子；顺式作用元件；基因表达；非生物胁迫

英文摘要： Our study showed that only 20% of the genes regulated by an ERF protein from Tamarix hispida contain GCC-box and DRE in their promoter regions, and similar results were also observed in studies of other ERF genes from T. hispida. Further studies showed that ERF may bind to other motif rather than GCC-box and DRE motifs to regulate gene expression in response to abiotic stresses. Based on this observation, further studies will be performed in this project. The ERF gene with high stress tolerance is transformed into T. hispida, and the transgenic T. hispida were treated with salt, drought, ABA or without treatment. After these treatments, ChIP libraries were respectively built from these samples, and ChIP-Seq were further performed to generate the DNA fragments bound by ERF under stress or normal growth conditions. The conserved motifs among these DNA fragments were identified, which may be recognized by ERF and will be further confirmed by yest one hybrid analysis. The frequencies of these motifs present in different ChIP libraries were analyezed to identify the motif bound by ERF with binding affinity in reponse to abiotic stress. The transcriptoms from wild type and transgenic T. hispida were built, and were assembled with the sequences generated from ChIP-Seq, and these motifs contained in which genes will be revealed by the assembled sequences. Based on these studies, the motifs recognzied by ERF protein and its binding affinities in response to abiotic stress will be identified and the stress tolerance pathways these motifs involved will be revealed.

英文关键词： Tamarix hispida;ERF transcription factor;cis-acting element;gene expression;abiotic stress