Abstract

Nitrogen (N) is an extremely important macronutrient for plant growth and development. It is the main limiting factor in most agricultural production. However, it is well known that the nitrogen use efficiency (NUE) of rice gradually decreases with the increase of the nitrogen application rate. In order to clarify the underlying metabolic and molecular mechanisms of this phenomenon, we performed an integrated analysis of the rice transcriptome and metabolome. Both differentially expressed genes (DEGs) and metabolite Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that carbon and nitrogen metabolism is significantly affected by nitrogen availability. Further analysis of carbon and nitrogen metabolism changes in rice under different nitrogen availability showed that high N inhibits nitrogen assimilation and aromatic metabolism pathways by regulating carbon metabolism pathways such as the tricarboxylic acid (TCA) cycle and the pentose phosphate pathway (PPP). Under low nitrogen, the TCA cycle is promoted to produce more energy and α-ketoglutarate, thereby enhancing nitrogen transport and assimilation. PPP is also inhibited by low N, which may be consistent with the lower NADPH demand under low nitrogen. Additionally, we performed a co-expression network analysis of genes and metabolites related to carbon and nitrogen metabolism. In total, 15 genes were identified as hub genes. In summary, this study reveals the influence of nitrogen levels on the regulation mechanisms for carbon and nitrogen metabolism in rice and provides new insights into coordinating carbon and nitrogen metabolism and improving nitrogen use efficiency in rice.

Highlights

  • Nitrogen is an extremely important and necessary macronutrient for plant growth and development [1]

  • The integrated analysis of the rice transcriptome and metabolome allowed us to get more insight into the carbon and nitrogen metabolism regulation mechanism in response to nitrogen availability, which could be of great practical significance in coordinating rice carbon and nitrogen metabolism and increasing nitrogen use efficiency (NUE)

  • Nitrogen deficiency had a negative effect on leaf biomass and leaf area, while high N promoted leaf biomass and leaf area

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Summary

Introduction

Nitrogen is an extremely important and necessary macronutrient for plant growth and development [1]. Nitrogen use efficiency (NUE) refers to the dry matter produced by plants that absorbs per unit mass of nitrogen and results from the coordination of plant carbon and nitrogen metabolism during development. Maintaining the coordination of carbon and nitrogen metabolism, and an appropriate balance of carbohydrates to nitrogen metabolites, referred to as the “C/N balance,” is important for plant growth, development, and yield production [6,8,17,18,19]. Reasonable regulation of carbon and nitrogen metabolism to improve NUE is an effective way to increase rice yield and reduce environmental costs [20,21]. The integrated analysis of the rice transcriptome and metabolome allowed us to get more insight into the carbon and nitrogen metabolism regulation mechanism in response to nitrogen availability, which could be of great practical significance in coordinating rice carbon and nitrogen metabolism and increasing NUE

Results
Metabolite Profiles of Rice Leaves in Response to Nitrogen Availability
Carbon Metabolism
Regulation of Differential Gene Expression by Transcription Factors
Discussion
Plant Materials and N Treatment
Analysis of Leaf Characteristics
RNA Extraction and Transcriptome Sequencing Analysis
Co-Expression Network Analysis of Genes and Metabolites
Confirmation of Transcriptome Data Using qRT-PCR Analysis
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