Engineering of red cells of Arabidopsis thaliana and comparative genome-wide gene expression analysis of red cells versus wild-type cells
Engineering of red cells of Arabidopsis thaliana and comparative genome-wide gene expression analysis of red cells versus wild-type cells
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DOI：
10.1007/s00425-010-1335-2
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发表时间：
2011-04-01
期刊：
影响因子：
4.3
通讯作者：
Xie, De-Yu
中科院分区：
文献类型：
--
作者：
Shi, Ming-Zhu;Xie, De-Yu
我们报道了拟南芥红细胞的代谢工程和与花青素生物合成相关的全基因组基因表达分析，以及红细胞和野生型（WT）细胞之间的其他代谢途径。本文首次从拟南芥花青素1-显性色素（pap1-D）产生的叶片中构建了拟南芥红细胞。这些红细胞产生7种花青素分子，其中包括一种通过LC-MS分析表征的新分子。作为对照的野生型细胞不产生花青素。全基因组微阵列分析显示，近66%和65%的基因组基因分别在红细胞和野生型细胞中表达。与WT细胞相比，红细胞中有3.2%的表达基因存在差异表达。红血球细胞中14个花青素生物合成途径相关基因的表达量明显高于WT细胞。基因芯片和RT-PCR分析表明TTG1-GL3/TT8-PAP1复合物调控花青素的生物合成。此外，红细胞与野生型细胞中表达水平差异显著的基因大多具有不同的生化功能，其中许多被定位于不同的代谢途径（如核糖体蛋白生物合成、光合作用、糖酵解、glyoxylate代谢和植物次生代谢）或细胞器（如叶绿体）。我们认为，两种细胞系之间基因表达谱的差异可能与细胞类型、PAP1的过表达以及花青素的高代谢通量有关。
We report metabolic engineering of Arabidopsis red cells and genome-wide gene expression analysis associated with anthocyanin biosynthesis and other metabolic pathways between red cells and wild-type (WT) cells. Red cells of A. thaliana were engineered for the first time from the leaves of production of anthocyanin pigment 1-Dominant (pap1-D). These red cells produced seven anthocyanin molecules including a new one that was characterized by LC-MS analysis. Wild-type cells established as a control did not produce anthocyanins. A genome-wide microarray analysis revealed that nearly 66 and 65% of genes in the genome were expressed in the red cells and wild-type cells, respectively. In comparison with the WT cells, 3.2% of expressed genes in the red cells were differentially expressed. The expression levels of 14 genes involved in the biosynthetic pathway of anthocyanin were significantly higher in the red cells than in the WT cells. Microarray and RT-PCR analyses demonstrated that the TTG1-GL3/TT8-PAP1 complex regulated the biosynthesis of anthocyanins. Furthermore, most of the genes with significant differential expression levels in the red cells versus the WT cells were characterized with diverse biochemical functions, many of which were mapped to different metabolic pathways (e.g., ribosomal protein biosynthesis, photosynthesis, glycolysis, glyoxylate metabolism, and plant secondary metabolisms) or organelles (e.g., chloroplast). We suggest that the difference in gene expression profiles between the two cell lines likely results from cell types, the overexpression of PAP1, and the high metabolic flux toward anthocyanins.