Comparative Multi-Omics Analysis of Broomcorn Millet in Response to Anthracocystis destruens Infection.

Anthracocystis destruens is the causal agent of broomcorn millet (Panicum miliaceum) smut disease, which results in serious yield losses in broomcorn millet production. However, the molecular basis underlying broomcorn millet defense against A. destruens is less understood. In this study, we investigated how broomcorn millet responds to infection by A. destruens by employing a comprehensive multi-omics approach. We examined the responses of broomcorn millet across transcriptome, metabolome, and microbiome levels. Infected leaves exhibited an upregulation of genes related to photosynthesis, accompanied by a higher accumulation of photosynthesis-related compounds and alterations in hormonal levels. However, broomcorn millet genes involved in immune response were downregulated post A. destruens infection, suggesting that A. destruens may suppress broomcorn millet immunity. In addition, we show that the immune suppression and altered host metabolism induced by A. destruens have no significant effect on the microbial community structure of broomcorn millet leaf, thus providing a new perspective for understanding the tripartite interaction between plant, pathogen, and microbiota.

Cadmium tolerance and accumulation from the perspective of metal ion absorption and root exudates in broomcorn millet.

Cadmium (Cd) is a persistent heavy metal that poses environmental and public health concerns. This study aimed to identify the potential biomarkers responsible for Cd tolerance and accumulation by investigating the response of the content of essential metal elements, transporter gene expression, and root exudates to Cd stress in broomcorn millet (Panicum miliaceum). A hydroponics experiment was conducted using two broomcorn millet cultivars with distinct Cd tolerance levels and accumulation phenotypes (Cd-tolerant and Cd-sensitive cultivars). Cd stress inhibited lateral root growth, especially in the Cd-sensitive cultivar. Furthermore, Cd accumulation was significantly greater in the Cd-tolerant cultivar than in the Cd-sensitive cultivar. Cd stress significantly inhibited the absorption of essential metal elements and significantly increased the calcium concentration. Differentially expressed genes involved in metal ion transport were identified via transcriptome analysis. Cd stress altered the composition of root exudates, thus increasing lipid species and decreasing alkaloid, lignan, sugar, and alcohol species. Moreover, Cd stress significantly reduced most alkaloid, organic acid, and phenolic acid exudates in the Cd-tolerant cultivar, while it increased most lipid and phenolic acid exudates in the Cd-sensitive cultivar. Some significantly changed root exudates (ferulic acid, O-coumaric acid, and spermine) are involved in the phenylalanine biosynthesis, and arginine and proline metabolic pathways, thus, may be potential biomarkers of Cd stress response. Overall, metal ion absorption and root exudates are critical for Cd tolerance and accumulation in broomcorn millet. These findings provide valuable insights into improving Cd phytoremediation by applying mineral elements or metabolites.

The Life Cycle and Ultrastructure of the Host Response of the Smut Pathogen Anthracocystis destruens on Broomcorn Millet.

Broomcorn millet smut caused by the fungus Anthracocystis destruens is one of the most destructive diseases in broomcorn millet production. The life cycle of A. destruens and host defense responses against A. destruens remain elusive. Here we investigated the disease symptom development and the parasitic process of A. destruens as well as the ultrastructure of the host-pathogen interface. The results showed that there are four typical symptoms of broomcorn millet smut, which are blackfly, cluster leaves, hedgehog head, and incomplete fruiting. A. destruens colonizes all tissues of broomcorn millet but produces teliospores only in the inflorescence. After infection, A. destruens proliferates in the host, likely in a systemic manner. Ultrastructural study of the infected inflorescence showed that the pathogen grows intercellularly and intracellularly within the host. The host activates defense response to prevent pathogen infection, accumulation of callose analogs and highly electron-dense deposits to resist A. destruens infection.

Exogenous 24-epibrassinolide boosts plant growth under alkaline stress from physiological and transcriptomic perspectives: The case of broomcorn millet (Panicum miliaceum L.).

Land alkalization is an abiotic stress that affects global sustainable agricultural development and the balance of natural ecosystems. In this study, two broomcorn millet cultivars, T289 (alkaline-tolerant) and S223 (alkaline-sensitive), were selected to investigate the response of broomcorn millet to alkaline stress and the role of brassinolide (BR) in alkaline tolerance. Phenotypes, physiologies, and transcriptomes of T289 and S223 plants under only alkaline stress (AS) and alkaline stress with BR (AB) were compared. The results showed that alkaline stress inhibited growth, promoted the accumulation of soluble sugars and malondialdehyde, enhanced electrolyte leakage, and destroyed the integrity of broomcorn millet stomata. In contrast, BR lessened the negative effects of alkaline stress on plants. Transcriptome sequencing analysis showed that relative to control groups (CK, nutrient solution), in AS groups, 21,113 and 12,151 differentially expressed genes (DEGs) were identified in S223 and T289, respectively. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed various terms and pathways related to metabolism. Compared to S223, alkaline stress strongly activated the brassinosteroid biosynthesis pathway in T289. Conversely, ARF, TF, and TCH4, associated with cell growth and elongation, were inhibited by alkaline stress in S223. Moreover, alkaline stress induced the activation of the mitogen-activated protein kinase (MAPK) pathway, the abscisic acid signaling pathway that initiates stomatal closure, as well as the starch and sucrose metabolism. The EG and BGL genes, which are associated with cellulose degradation, were notably activated. BR enhanced alkaline tolerance, thereby alleviating the transcriptional responses of the two cultivars. Cultivar T289 is better in alkalized regions. Taken together, these results reveal how broomcorn millet responds to alkaline stress and BR mitigates alkaline stress, thus promoting agriculture in alkalized regions.

Endogenous bioactive gibberellin/abscisic acids and enzyme activity synergistically promote the phytoremediation of alkaline soil by broomcorn millet (Panicum miliaceum L.).

Broomcorn millet (Panicum miliaceum L.), an important food crop, grows in arid and semi-arid areas that face soil saline-alkalization. To date, no studies have investigated the mechanisms by which broomcorn millet seeds respond to and tolerate alkali stress. In this study, six broomcorn millet genotypes (B102, B220, B269, B279, B289, and B297) were selected to explore the physiological and molecular mechanisms of alkali stress at the germination stage. The results showed that alkali stress delayed the germination of broomcorn millet, and α-amylase activity was positively correlated with the germination rate. After alkali stress, the genotypes with lower alkali damage rates exhibited stronger antioxidant defenses. Real-time polymerase chain reaction analysis showed that alkali stress downregulated gibberellic acid (GA) synthesis genes but upregulated GA inactivation and abscisic acid (ABA) synthesis genes. Similarly, seeds displayed lower GA concentrations and higher ABA concentrations after alkali stress. Therefore, the ratios of various GAs/ABA decreased within the range of 35.77% to approximately 96.45%. Additionally, genotypes associated with lower alkali damage rates had higher GA/ABA ratios. These findings indicate that the alkali tolerance of broomcorn millet at the germination stage may be attributed to higher GA/ABA ratios, higher α-amylase activity, and stronger antioxidant defense, which synergistically resist alkali stress. This study will contribute to molecular breeding aiming to enhance alkali-tolerance and restoration of alkaline soils.

Leaf Transcriptome Analysis of Broomcorn Millet Uncovers Key Genes and Pathways in Response to Sporisorium destruens.

Broomcorn millet (Panicum miliaceum L.) affected by smut (caused by the pathogen Sporisorium destruens) has reduced production yields and quality. Determining the tolerance of broomcorn millet varieties is essential for smut control. This study focuses on the differences in the phenotypes, physiological characteristics, and transcriptomes of resistant and susceptible broomcorn millet varieties under Sporisorium destruens stress. In diseased broomcorn millet, the plant height and stem diameter were reduced, while the number of nodes increased. After infection, the activities of superoxide dismutase and peroxidase decreased, and malondialdehyde and relative chlorophyll content (SPAD) decreased. Transcriptome analysis showed 514 and 5452 differentially expressed genes (DEGs) in the resistant and susceptible varieties, respectively. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs showed that pathways related to plant disease resistance, such as phenylpropanoid biosynthesis, plant-pathogen interaction, and plant hormone signal transduction, were significantly enriched. In addition, the transcriptome changes of cluster leaves and normal leaves in diseased broomcorn millet were analysed. Gene ontology and KEGG enrichment analyses indicated that photosynthesis played an important role in both varieties. These findings lay a foundation for future research on the molecular mechanism of the interaction between broomcorn millet and Sporisorium destruens.

Widely targeted metabolomic analysis revealed the effects of alkaline stress on nonvolatile and volatile metabolites in broomcorn millet grains.

Broomcorn millet (BM) is a future smart food. However, no information is available on the metabolism of BM grains under alkaline stress. In this study, the effects of alkaline stress on nonvolatile and volatile metabolites in the BM grains of two varieties (S223 and T289) were investigated through metabolomics approaches. All 933 nonvolatile metabolites and 313 volatile metabolites were identified, with 114 and 89 nonvolatile metabolites and 16 and 20 volatile metabolites accumulating differentially under normal vs. alkaline stresses of S223 and T289, respectively. The results indicated that alkaline stress altered phenylpropanoids, flavonoids, flavone and flavonol, valine, leucine, and isoleucine biosynthesis, as well as arginine, proline, tryptophan, and ascorbate metabolism. The effects of alkaline stress were not identical between the two varieties, which could lead to variations in active substance content. These results provide valuable information for further studies on food chemistry and the functional food development of BM grains.

Comparison of the fine structure and physicochemical properties of proso millet (Panicum miliaceum L.) starch from different ecological regions.

Field experiments were conducted to evaluate the morphology, granule size, fine structure, thermal properties, and pasting properties of starch from a waxy (139) and a non-waxy (297) varieties of proso millet grown in Yulin (YY) and Yangling (YL). Compared with the starches from the two varieties grown in YY, the starches from the two varieties grown in YL exhibited higher relative crystallinities, 1045/1022 cm-1 ratio, and amounts of amylopectin long branch chains (APL) but lower 1022/995 cm -1 ratio, amounts of amylopectin short branch chains (APs), and APs/APL ratios. Starches from YL also synthesized long branch-chain amylopectin to enhance intermolecular interactions and form a stable granular structure, which resulted in increased starch gelatinization temperature, enhanced shear resistance, and reduced setback viscosity. Starch from the waxy (139) variety has good application prospects in the food industry because of its high gelatinization temperature and light transmittance and low setback value, which can be ascribed to its extremely low amylose content, polydispersity index, high molecular weight, and dispersed molecular density. It may serve as a reference for applying proso millet starches in the food industry and developing breeding programs to improve starch quality.

Nitrogen fertilizer affects starch synthesis to define non-waxy and waxy proso millet quality.

Understanding the effect of nitrogen fertilization on the quality of proso millet is key to expanding the use of this crop to address water scarcity and food security. Therefore, this study determined the impact of nitrogen fertilization on the proso millet quality. Nitrogen fertilization significantly increased the NR and GS activities and decreased the GBSSase activity, resulting in an increase in protein content and reduction in amylose content and L*, which decreased the appearance quality. Nitrogen fertilization increased the proportion of short amylopectin chains, resulting in a more disordered carbohydrate structure, and decreased the proportion of hydrophilic functional groups, contributing to an increase in setback viscosity and decrease in pasting temperature in the waxy (w139) variety. In contrast, the non-waxy (n297) variety exhibited a larger proportion of long amylopectin chains, lower ordered structure and hydrophobic functional groups after nitrogen fertilization, which strengthened the inter- and intramolecular forces of starch colloids.

[Determination of N,N'-ethylenediamine disuccinic acid in bioconversion reaction solution using high performance liquid chromatography].

A method was developed for the determination of N,N'-ethylenediamine disuccinic acid (EDDS) in bioconversion samples by high performance liquid chromatography (HPLC). An InertSustain AQ-C18 column (250 mm×4.6 mm, 5 µm) was used in the analysis. The mobile phase was 25% (v/v) methanol with 1.0 g/L Cu(CO2CH3)2·H2O, 2.0 g/L tetrabutylammonium hydroxide, and the pH was adjusted to 2.80 with phosphoric acid. The flow rate of the mobile phase was 1.0 mL/min, and the column temperature was set at 30℃. The detection wavelength was 254 nm. EDDS, fumaric acid, citric acid, malic acid and ethylenediaminetetraacetic acid (EDTA) were separated from one another within 8 min. EDDS showed good linearity in the range of 0.06-0.6 g/L. About 0.25 g/L EDDS was detected in the biosynthesis reaction solution catalyzed by recombinant EDDS-lyase, while 36.56 g/L malic acid was formed as the by-product from fumaric acid. The catalytic activity of the enzyme was confirmed in the hydrolysis of 10 g/L EDDS, which produced 3.05 g/L malic acid in 3 h. This analytical method is simple, rapid, sensitive, reliable, and suitable for the analysis in the research of EDDS bioconversion process.