ABSTRACT
KEY MESSAGE: The study established split-root system (SRS) in foxtail millet, and identified the molecular regulatory mechanisms and metabolic pathways related to systemic nitrogen signaling based on this system and transcriptome analysis. The growth of crops is primarily constrained by the availability of nitrogen (N), an essential nutrient. Foxtail millet (Setaria italica L.) is a significant orphan crop known for its strong tolerance to barren conditions. Despite this, the signaling pathway of nitrogen in foxtail millet remains largely unexplored. Identifying the candidate genes responsible for nitrogen response in foxtail millet is crucial for enhancing its agricultural productivity. This study utilized the split-root system (SRS) in foxtail millet to uncover genes associated with Systemic Nitrogen Signaling (SNS). Transcriptome analysis of the SRS revealed 2158 differentially expressed genes (DEGs) implicated in SNS, including those involved in cytokinin synthesis, transcription factors, E3 ubiquitin ligase, and ROS metabolism. Silencing of SiIPT5 and SiATL31 genes through RNAi in transgenic plants resulted in reduced SNS response, indicating their role in the nitrogen signaling pathway of foxtail millet. Furthermore, the induction of ROS metabolism-related genes in response to KNO3 of the split-root System (Sp.KNO3) suggests a potential involvement of ROS signaling in the SNS of foxtail millet. Overall, this study sheds light on the molecular regulatory mechanisms and metabolic pathways of foxtail millet in relation to SNS.
Subject(s)
Gene Expression Profiling , Gene Expression Regulation, Plant , Nitrogen , Plant Roots , Setaria Plant , Signal Transduction , Setaria Plant/genetics , Setaria Plant/metabolism , Nitrogen/metabolism , Plant Roots/genetics , Plant Roots/metabolism , Signal Transduction/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified , Transcriptome/genetics , Reactive Oxygen Species/metabolismABSTRACT
Amino acid permeases (AAPs) transporters are crucial for the long-distance transport of amino acids in plants, from source to sink. While Arabidopsis and rice have been extensively studied, research on foxtail millet is limited. This study identified two transcripts of SiAAP9, both of which were induced by NO3- and showed similar expression patterns. The overexpression of SiAAP9L and SiAAP9S in Arabidopsis inhibited plant growth and seed size, although SiAAP9 was found to transport more amino acids into seeds. Furthermore, SiAAP9-OX transgenic Arabidopsis showed increased tolerance to high concentrations of glutamate (Glu) and histidine (His). The high overexpression level of SiAAP9 suggested its protein was not only located on the plasma membrane but potentially on other organelles, as well. Interestingly, sequence deletion reduced SiAAP9's sensitivity to Brefeldin A (BFA), and SiAAP9 had ectopic localization on the endoplasmic reticulum (ER). Protoplast amino acid uptake experiments indicated that SiAAP9 enhanced Glu transport into foxtail millet cells. Overall, the two transcripts of SiAAP9 have similar functions, but SiAAP9L shows a higher colocalization with BFA compartments compared to SiAAP9S. Our research identifies a potential candidate gene for enhancing the nutritional quality of foxtail millet through breeding.