As one of the oldest agricultural crops in China, millet (Panicum miliaceum) has powerful drought tolerance. In this study, transcriptome and metabolome analyses of ‘Hequ Red millet’ (HQ) and ‘Yanshu No.10’ (YS10) millet after 6 h of drought stress were performed. Transcriptome characteristics of drought stress in HQ and YS10 were characterized by Pacbio full-length transcriptome sequencing. The pathway analysis of the differentially expressed genes (DEGs) showed that the highly enriched categories were related to starch and sucrose metabolism, pyruvate metabolism, metabolic pathways, and the biosynthesis of secondary metabolites when the two millet varieties were subjected to drought stress. Under drought stress, 245 genes related to energy metabolism were found to show significant changes between the two strains. Further analysis showed that 219 genes related to plant hormone signal transduction also participated in the drought response. In addition, numerous genes involved in anthocyanin metabolism and photosynthesis were confirmed to be related to drought stress, and these genes showed significant differential expression and played an important role in anthocyanin metabolism and photosynthesis. Moreover, we identified 496 transcription factors related to drought stress, which came from 10 different transcription factor families, such as bHLH, C3H, MYB, and WRKY. Further analysis showed that many key genes related to energy metabolism, such as citrate synthase, isocitrate dehydrogenase, and ATP synthase, showed significant upregulation, and most of the structural genes involved in anthocyanin biosynthesis also showed significant upregulation in both strains. Most genes related to plant hormone signal transduction showed upregulated expression, while many JA and SA signaling pathway-related genes were downregulated. Metabolome analysis was performed on ‘Hequ red millet’ (HQ) and ‘Yanshu 10’ (YS10), a total of 2082 differential metabolites (DEMs) were identified. These findings indicate that energy metabolism, anthocyanins, photosynthesis, and plant hormones are closely related to the drought resistance of millet and adapt to adversity by precisely regulating the levels of various molecular pathways.

The flowering stage is a critical period for water sensitivity and quality formation of broomcorn millets. However, the effects and mechanisms of drought stress on the quality formation of broomcorn millets are not clear. We used the drought-resistant variety Hequ red millet (H) and the drought-sensitive variety Yanshu No. 10 (Y) were used as materials for drought stress treatment during the flowering stage, metabolomics and physiological methods were used to study the differences in protein, starch, amino acids, medium and medium-long chain fatty acids, and their response characteristics to drought in broomcorn millet. The results showed that different genotypes of broomcorn millets exhibited different response mechanisms in the face of drought stress. In Hequ red millet, drought stress significantly increased the contents of amylopectin (2.57%), pyridoxine (31.89%), and anthocyanin, and significantly decreased the contents of water-soluble protein (5.82%), glutelin (10.07%), thiamine (14.95%) and nicotinamide (23.01%). In Yanshu No. 10, drought significantly decreased amylose by 6.05%, and significantly increased riboflavin and nicotinamide contents by 21.11% and 32.59%. Correlation analysis showed that total starch and amylose were highly significantly positively correlated with methyl palmitate; negatively correlated with amylopectin, vitamins, proteins, free amino acids, and medium-long chain fatty acids; and amylopectin was significantly positively correlated with water-soluble protein, riboflavin, and pyridoxine. Water-soluble protein and glutelin were significantly positively correlated with most free amino acids, medium-long chain fatty acids, and nicotinamide. Thiamine showed significant positive correlation with nicotinamide and significant negative correlation with pyridoxine. Riboflavin was significantly positively correlated with nicotinamide, pyridoxine, and water-soluble protein, and pyridoxine was significantly positively correlated with water-soluble protein. Hequ red millet transforms into amylopectin by consuming water-soluble protein and glutelin, and improves drought resistance by accumulating pyridoxine, and changes its physicochemical properties by decreasing the content of amylose and protein and elevating the content of amylopectin. Yanshu No. 10 resisted drought by catabolizing lipids to produce fatty acids and by consuming amylose for conversion into other metabolites. The present study helps to understand the response of the nutritional quality of millets to drought stress at the flowering stage and provides a theoretical basis for the selection and breeding of superior varieties of millets and drought resistance research.

In order to clarify the effect of different fertilizers on foxtail millet quality under low nitrogen conditions, we used JGNo.21 and LZGNo.2 as experimental materials and set up five treatments, including non-fertilization, nitrogen, phosphorus, compound, and organic fertilizers, to study the regulation of different fertilizer types on agronomic traits, nutrient fractions, and pasting characteristics of foxtail millet under low nitrogen conditions. Compared with the control, all of the fertilizers improved the agronomic traits of JGNo.21 to a certain extent. Nitrogen and compound fertilizer treatments reduced the starch content of JGNo.21; the starch content was reduced by 0.55% and 0.07% under nitrogen and compound fertilizers treatments. Phosphorus and organic fertilizers increased starch content, and starch content increased by 0.50% and 0.56% under phosphorus and organic fertilizer treatments. The effect of each fertilizer treatment on protein content was completely opposite to that of starch; different fertilizer treatments reduced the fat content of JGNo.21 and increased the fiber content. Among them, nitrogen and phosphorus fertilizers increased the yellow pigment content; the yellow pigment content increased by 1.21% and 2.64% under nitrogen and phosphorus fertilizer treatments. Organic and compound fertilizers reduced the content of yellow pigment; the yellow pigment content was reduced by 3.36% and 2.79% under organic and compound fertilizer treatments. Nitrogen and organic fertilizers increased the fat content of LZGNo.2; the fat content increased by 2.62% and 1.98% under nitrogen, organic fertilizer treatment. Compound and phosphorus fertilizer decreased the fat content; the fat content decreased by 2.16% and 2.90% under compound and phosphorus fertilizer treatment. Different fertilizer treatments reduced the cellulose and yellow pigment content of LZGNo.2. The content of essential, non-essential, and total amino acids of JGNo.21 was increased under compound and nitrogen fertilizer treatments and decreased under organic and phosphorus fertilizer treatments. The content of essential, non-essential, and total amino acids of LZGNo.2 was significantly higher under compound, nitrogen, and organic fertilizer treatments compared with control and significantly decreased under phosphorus fertilizer treatments. Nitrogen and compound fertilizer treatments significantly reduced the values of peak viscosity, trough viscosity, breakdown viscosity, final viscosity, setback viscosity, and pasting time of each index of JGNo.21; phosphorus and organic fertilizer treatments improved the values of each index. In contrast, the pasting viscosity of LZGNo.2 increased under phosphorus fertilizer treatment and decreased under nitrogen fertilizer treatment. Reasonable fertilization can improve the quality of foxtail millet, which provides a scientific theoretical basis for improving the quality of foxtail millet.

Drought stress is an important abiotic stress factor restricting crop production. Broomcorn millet (Panicum miliaceum L.) has become an ideal material for analyzing the stress adaptation mechanisms of crops due to its strong stress resistance. However, the functional characteristics of its rhizosphere microorganisms in response to drought remain unclear. In this study, metagenomics and metabolomics techniques were employed to systematically analyze the compositional characteristics of the microbial community, functional properties, and changes in metabolites in the rhizosphere soil of broomcorn millet under drought stress. On this basis, an analysis was conducted in combination with the differences in functional pathways. The results showed that the drought treatment during the flowering stage significantly altered the species composition of the rhizosphere microorganisms of broomcorn millet. Among them, the relative abundances of beneficial microorganisms such as Nitrosospira, Coniochaeta, Diversispora, Gigaspora, Glomus, and Rhizophagus increased significantly. Drought stress significantly affects the metabolic pathways of rhizosphere microorganisms. The relative abundances of genes associated with prokaryotes, glycolysis/gluconeogenesis, and other metabolic process (e.g., ribosome biosynthesis, amino sugar and nucleotide sugar metabolism, and fructose and mannose metabolism) increased significantly. Additionally, the expression levels of functional genes involved in the phosphorus cycle were markedly upregulated. Drought stress also significantly alters the content of specific rhizosphere soil metabolites (e.g., trehalose, proline). Under drought conditions, broomcorn millet may stabilize the rhizosphere microbial community by inducing its restructuring and recruiting beneficial fungal groups. These community-level changes can enhance element cycling efficiency, optimize symbiotic interactions between broomcorn millet and rhizosphere microorganisms, and ultimately improve the crop’s drought adaptability. Furthermore, the soil metabolome (e.g., trehalose and proline) functions as a pivotal interfacial mediator, orchestrating the interaction network between broomcorn millet and rhizosphere microorganisms, thereby enhancing plant stress tolerance. This study sheds new light on the functional traits of rhizosphere microbiota under drought stress and their mechanistic interactions with host plants.

A water-soluble polysaccharide from foxtail millet (FM-D1) was isolated and purified through gradient ethanol precipitation and column chromatography. Size-Exclusion Chromatography–Multi-Angle Light Scattering–Refractive Index (SEC-MALLS-RI) and high-performance anion-exchange chromatography (HPAEC) analyses revealed that FM-D1 constitutes a highly purified neutral polysaccharide exclusively composed of glucose as the sugar unit, with a molecular weight of 14.823 kDa. The structural characterization results obtained from gas chromatography–mass spectrometer (GC–MS) and nuclear magnetic resonance spectroscopy (NMR) spectra suggest that FM-D1 primarily consists of a main chain linked by →4)-α-D-Glcp-(1→ and minor quantities of →4,6)-α-D-Glcp-(1→ to form the main chain, with branching mainly composed of α-D-Glcp-(1→ attached to the O-6 position of →4,6)-α-D-Glcp-(1→ sugar residues. Based on these findings, the antioxidant and immunomodulatory activities of FM-D1 were evaluated in vitro. The results indicated that FM-D1 exhibited moderate 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radical scavenging capacity and total antioxidant capacity (TAOC). Furthermore, FM-D1 stimulated macrophage proliferation and inhibited the production of nitric oxide (NO) and inflammatory factors (TNF-α, IL-1β, and IL-6) in lipopolysaccharide (LPS)-stimulated RAW 246.7 cells. Overall, the findings of this study suggest that foxtail millet holds promise as a potential antioxidant agent and immunologic substance in functional foods.