Lotus japonicus metabolic profiling.: Development of gas chromatography-mass spectrometry resources for the study of plant-microbe interactions
Lotus japonicus metabolic profiling.: Development of gas chromatography-mass spectrometry resources for the study of plant-microbe interactions
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DOI：
10.1104/pp.104.054957
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发表时间：
2005-04-01
期刊：
影响因子：
7.4
通讯作者：
Udvardi, MK
中科院分区：
文献类型：
--
作者：
Desbrosses, GG;Kopka, J;Udvardi, MK
豆科植物根瘤的共生固氮需要植物和细菌代谢的分化和整合。生物化学、分子生物学和遗传学的经典方法已经揭示了豆科植物根瘤的初级代谢的许多方面，这些方面是SNF的基础。功能基因组学方法，特别是转录组学和蛋白质组学，开始为豆科植物根瘤的代谢潜力提供更全面的了解，如紫花苜蓿和日本莲花。为了扩展这些方法，我们建立了使用气相色谱-质谱联用的无偏测量和分析日本草数百种代谢物的方案。在建立了代表已知和未知代谢物的质谱标记库之后，我们测量并比较了共生植物根瘤、根、叶和花的相对代谢物水平。数据的主成分分析揭示了不同器官的不同代谢表型，并导致每个器官的标记代谢物的鉴定。结节中富集的代谢物包括：十八烷酸、天冬酰胺、谷氨酸、同型丝氨酸、半胱氨酸、腐胺、甘露醇、苏氨酸、葡萄糖酸、甘油三酯和甘油3-磷酸。根据荷花不同器官的分布模式，分层聚类分析可以区分出10组代谢物。本文所描述的资源和工具，以及目前正在进行的基因组测序、L. japonicus和Mesorhizobitm loti的转录组和蛋白质组分析，将有助于更好地理解SNE基础上的结节代谢
Symbiotic nitrogen fixation (SNF) in legume root nodules requires differentiation and integration of both plant and bacterial metabolism. Classical approaches of biochemistry, molecular biology, and genetics have revealed many aspects of primary metabolism in legume nodules that underpin SNF Functional genomics approaches, especially transcriptomics and proteomics, are beginning to provide a more holistic picture of the metabolic potential of nodules in model legumes like Medicago truncatula and Lotus japonicus. To extend these approaches, we have established protocols for nonbiased measurement and analysis of hundreds of metabolites from L. japonicus, using gas chromatography coupled with mass spectrometry. Following creation of mass spectral tag libraries, which represent both known and unknown metabolites, we measured and compared relative metabolite levels in nodules, roots, leaves, and flowers of symbiotic plants. Principal component analysis of the data revealed distinct metabolic phenotypes for the different organs and led to the identification of marker metabolites for each. Metabolites that were enriched in nodules included: octadecanoic acid, asparagine, glutamate, homoserine, cysteine, putrescine, mannitol, threonic acid, gluconic acid, glyceric acid-3-P, and glycerol-3-P. Hierarchical cluster analysis enabled discrimination of 10 groups of metabolites, based on distribution patterns in diverse Lotus organs. The resources and tools described here, together with ongoing efforts in the areas of genome sequencing, and transcriptome and proteome analysis of L. japonicus and Mesorhizobitm loti, should lead to a better understanding of nodule metabolism that underpins SNE