Concurrent isotope-assisted metabolic flux analysis and transcriptome profiling reveal responses of poplar cells to altered nitrogen and carbon supply
| Autor: | Xiaofeng Zhang, Ashish Misra, Shilpa Nargund, Ganesh Sriram, Gary D. Coleman |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Nitrogen Citric Acid Cycle Plant Science Pentose phosphate pathway Biology Photosynthesis 01 natural sciences Pentose Phosphate Pathway Transcriptome 03 medical and health sciences Nutrient Acetyl Coenzyme A Gene Expression Regulation Plant Metabolic flux analysis Botany Genetics Carbon Isotopes Gene Expression Profiling Cell Biology Carbon Metabolic Flux Analysis Citric acid cycle Metabolic pathway Populus 030104 developmental biology Biochemistry Plant nutrition 010606 plant biology & botany |
| Zdroj: | The Plant Journal. 93:472-488 |
| ISSN: | 0960-7412 |
| Popis: | Reduced nitrogen is indispensable to plants. However, its limited availability in soil combined with the energetic and environmental impacts of nitrogen fertilizers motivates research into molecular mechanisms toward improving plant nitrogen use efficiency (NUE). We performed a systems-level investigation of this problem by employing multiple 'omics methodologies on cell suspensions of hybrid poplar (Populus tremula × Populus alba). Acclimation and growth of the cell suspensions in four nutrient regimes ranging from abundant to deficient supplies of carbon and nitrogen revealed that cell growth under low-nitrogen levels was associated with substantially higher NUE. To investigate the underlying metabolic and molecular mechanisms, we concurrently performed steady-state 13 C metabolic flux analysis with multiple isotope labels and transcriptomic profiling with cDNA microarrays. The 13 C flux analysis revealed that the absolute flux through the oxidative pentose phosphate pathway (oxPPP) was substantially lower (~threefold) under low-nitrogen conditions. Additionally, the flux partitioning ratio between the tricarboxylic acid cycle and anaplerotic pathways varied from 84%:16% under abundant carbon and nitrogen to 55%:45% under deficient carbon and nitrogen. Gene expression data, together with the flux results, suggested a plastidic localization of the oxPPP as well as transcriptional regulation of certain metabolic branchpoints, including those between glycolysis and the oxPPP. The transcriptome data also indicated that NUE-improving mechanisms may involve a redirection of excess carbon to aromatic metabolic pathways and extensive downregulation of potentially redundant genes (in these heterotrophic cells) that encode photosynthetic and light-harvesting proteins, suggesting the recruitment of these proteins as nitrogen sinks in nitrogen-abundant conditions. |
| Databáze: | OpenAIRE |
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