Max-Planck Institute of Molecular Plant Physiology, Am Muehlenberg 1, D-14476 Potsdam, Germany.
Plant J. 2011 Dec;68(6):999-1013. doi: 10.1111/j.1365-313X.2011.04750.x. Epub 2011 Oct 25.
Maturation of fleshy fruits such as tomato (Solanum lycopersicum) is subject to tight genetic control. Here we describe the development of a quantitative real-time PCR platform that allows accurate quantification of the expression level of approximately 1000 tomato transcription factors. In addition to utilizing this novel approach, we performed cDNA microarray analysis and metabolite profiling of primary and secondary metabolites using GC-MS and LC-MS, respectively. We applied these platforms to pericarp material harvested throughout fruit development, studying both wild-type Solanum lycopersicum cv. Ailsa Craig and the hp1 mutant. This mutant is functionally deficient in the tomato homologue of the negative regulator of the light signal transduction gene DDB1 from Arabidopsis, and is furthermore characterized by dramatically increased pigment and phenolic contents. We choose this particular mutant as it had previously been shown to have dramatic alterations in the content of several important fruit metabolites but relatively little impact on other ripening phenotypes. The combined dataset was mined in order to identify metabolites that were under the control of these transcription factors, and, where possible, the respective transcriptional regulation underlying this control. The results are discussed in terms of both programmed fruit ripening and development and the transcriptional and metabolic shifts that occur in parallel during these processes.
肉质果实(如番茄)的成熟受严格的遗传控制。在这里,我们描述了一种定量实时 PCR 平台的开发,该平台可准确量化大约 1000 个番茄转录因子的表达水平。除了利用这种新方法,我们还使用 GC-MS 和 LC-MS 分别进行了 cDNA 微阵列分析和初级和次级代谢物的代谢物分析。我们将这些平台应用于整个果实发育过程中采集的果皮材料,研究了野生型番茄 Solanum lycopersicum cv. Ailsa Craig 和 hp1 突变体。该突变体在番茄中功能缺失了拟南芥光信号转导基因 DDB1 的负调控因子的同源物,并且特征在于色素和酚类物质含量显著增加。我们选择这个特定的突变体是因为它之前已经显示出在几种重要果实代谢物的含量方面有显著变化,但对其他成熟表型的影响相对较小。对合并数据集进行了挖掘,以确定受这些转录因子控制的代谢物,并且在可能的情况下,确定这种控制背后的相应转录调节。结果从编程果实成熟和发育的角度以及在这些过程中同时发生的转录和代谢变化进行了讨论。