Roessner U, Willmitzer L, Fernie A R
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Golm, Germany.
Plant Physiol. 2001 Nov;127(3):749-64.
We conducted a comprehensive metabolic phenotyping of potato (Solanum tuberosum L. cv Desiree) tuber tissue that had been modified either by transgenesis or exposure to different environmental conditions using a recently developed gas chromatography-mass spectrometry profiling protocol. Applying this technique, we were able to identify and quantify the major constituent metabolites of the potato tuber within a single chromatographic run. The plant systems that we selected to profile were tuber discs incubated in varying concentrations of fructose, sucrose, and mannitol and transgenic plants impaired in their starch biosynthesis. The resultant profiles were then compared, first at the level of individual metabolites and then using the statistical tools hierarchical cluster analysis and principal component analysis. These tools allowed us to assign clusters to the individual plant systems and to determine relative distances between these clusters; furthermore, analyzing the loadings of these analyses enabled identification of the most important metabolites in the definition of these clusters. The metabolic profiles of the sugar-fed discs were dramatically different from the wild-type steady-state values. When these profiles were compared with one another and also with those we assessed in previous studies, however, we were able to evaluate potential phenocopies. These comparisons highlight the importance of such an approach in the functional and qualitative assessment of diverse systems to gain insights into important mediators of metabolism.
我们使用最近开发的气相色谱 - 质谱分析方法，对经过转基因或暴露于不同环境条件处理的马铃薯（Solanum tuberosum L. cv Desiree）块茎组织进行了全面的代谢表型分析。应用该技术，我们能够在一次色谱运行中鉴定和定量马铃薯块茎的主要组成代谢物。我们选择进行分析的植物系统包括在不同浓度的果糖、蔗糖和甘露醇中培养的块茎切片以及淀粉生物合成受损的转基因植物。然后，首先在单个代谢物水平上，接着使用层次聚类分析和主成分分析等统计工具对所得图谱进行比较。这些工具使我们能够将聚类分配给各个植物系统，并确定这些聚类之间的相对距离；此外，分析这些分析的载荷能够识别这些聚类定义中最重要的代谢物。用糖处理过的切片的代谢图谱与野生型稳态值有显著差异。然而，当将这些图谱相互比较以及与我们在先前研究中评估的图谱进行比较时，我们能够评估潜在的拟表型。这些比较突出了这种方法在对不同系统进行功能和定性评估以深入了解代谢重要调节因子方面的重要性。
