Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor
Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor
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DOI：
10.1111/j.1365-313x.2005.02371.x
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发表时间：
2005-04-01
期刊：
影响因子：
7.2
通讯作者：
Saito, K
中科院分区：
文献类型：
--
作者：
Tohge, T;Nishiyama, Y;Saito, K
代谢组学和转录组学的整合可以提供基因-代谢物网络的精确信息，用于鉴定未知基因的功能，除非存在转录后修饰。在这里，我们报告了一个全面的分析代谢组和转录组的拟南芥过表达的PAP 1基因编码的MYB转录因子，识别新的基因功能参与类黄酮的生物合成。对于代谢组学分析，我们通过高效液相色谱-质谱法进行类黄酮靶向分析，并通过具有超高分辨率能力的傅立叶变换离子回旋质谱法进行非靶向分析。这种组合分析揭示了花青素和槲皮素衍生物的特异性积累，并从PAP 1过表达植物中推定的1800种代谢物中鉴定出8种新型花青素。对22810个基因的转录组分析显示，38个基因的诱导异位PAP 1过表达。除了已知的参与花青素苷产生的基因外，还发现了几个功能未知或被认为具有功能的基因，包括糖基转移酶、酰基转移酶、谷胱甘肽S-转移酶、糖转运蛋白和转录因子等。两个推定的糖基转移酶基因（At 5g 17050和At 4g 14090）诱导的PAP 1表达被证实分别编码类黄酮3-O-葡萄糖基转移酶和花青素5-O-葡萄糖基转移酶，从它们的重组蛋白在体外的酶活性和在各自的T-DNA插入突变体中的花青素的分析结果。功能基因组学的方法，通过整合代谢组学和转录组学在这里提供了一个创新的手段，确定新的基因功能参与植物代谢。
The integration of metabolomics and transcriptomics can provide precise information on gene-to-metabolite networks for identifying the function of unknown genes unless there has been a post-transcriptional modification. Here, we report a comprehensive analysis of the metabolome and transcriptome of Arabidopsis thaliana over-expressing the PAP1 gene encoding an MYB transcription factor, for the identification of novel gene functions involved in flavonoid biosynthesis. For metabolome analysis, we performed flavonoid-targeted analysis by high-performance liquid chromatography-mass spectrometry and non-targeted analysis by Fourier-transform ion-cyclotron mass spectrometry with an ultrahigh-resolution capacity, This combined analysis revealed the specific accumulation of cyanidin and quercetin derivatives, and identified eight novel anthocyanins from an array of putative 1800 metabolites in PAP1 over-expressing plants. The transcriptome analysis of 22 810 genes on a DNA microarray revealed the induction of 38 genes by ectopic PAP1 overexpression. In addition to well-known genes involved in anthocyanin production, several genes with unidentified functions or annotated with putative functions, encoding putative glycosyltransferase, acyltransferase, glutathione S-transferase, sugar transporters and transcription factors, wore induced by PAP1. Two putative glycosyltransferase genes (At5g17050 and At4g14090) induced by PAP1 expression were confirmed to encode flavonoid 3-O-glucosyltransferase and anthocyanin 5-O-glucosyltransferase, respectively, from the enzymatic activity of their recombinant proteins in vitro and results of the analysis of anthocyanins in the respective T-DNA-inserted mutants. The functional genomics approach through the integration of metabolomics and transcriptomics presented here provides an innovative means of identifying novel gene functions involved in plant metabolism.