Glutathione S-transferases (GSTs) are a critical superfamily of multifunctional enzymes in plants. As a ligand or binding protein, GSTs regulate plant growth and development and detoxification. Foxtail millet (Setaria italica (L.) P. Beauv) could respond to abiotic stresses through a highly complex multi-gene regulatory network in which the GST family is also involved. However, GST genes have been scarcely studied in foxtail millet. Genome-wide identification and expression characteristics analysis of the foxtail millet GST gene family were conducted by biological information technology. The results showed that 73 GST genes (SiGSTs) were identified in the foxtail millet genome and were divided into seven classes. The chromosome localization results showed uneven distribution of GSTs on the seven chromosomes. There were 30 tandem duplication gene pairs belonging to 11 clusters. Only one pair of SiGSTU1 and SiGSTU23 were identified as fragment duplication genes. A total of ten conserved motifs were identified in the GST family of foxtail millet. The gene structure of SiGSTs is relatively conservative, but the number and length of exons of each gene are still different. The cis-acting elements in the promoter region of 73 SiGST genes showed that 94.5% of SiGST genes possessed defense and stress-responsive elements. The expression profiles of 37 SiGST genes covering 21 tissues suggested that most SiGST genes were expressed in multiple organs and were highly expressed in roots and leaves. By qPCR analysis, we found that 21 SiGST genes were responsive to abiotic stresses and abscisic acid (ABA). Taken together, this study provides a theoretical basis for identifying foxtail millet GST family information and improving their responses to different stresses.

Ear photosynthesis plays a key role in wheat photosynthesis during the grain filling stage, particularly under drought stress. Thus, dissecting the responsibilities of the glume and awn in photosynthetic carbon fixation and assimilates transportation during the grain filling stage in spikes is imperative. In this study, the detachment of the glume (DG) and awn (DA) of a wheat variety (Pubing143) was used to estimate their influences on ear photosynthesis and dry matter distribution. Radioactive carbon-14 (¹⁴C) isotope was detected by a multifunctional liquid scintillation counting system. The accumulation of ¹⁴C assimilates and their contributions to grain weight were then calculated. Under well-watered conditions, ear photosynthesis was reduced by 16.8 % and 46.2 % 25 d after anthesis (DAA) in the de-glumed control (DGC) and de-awned control (DAC) treatments, respectively, compared with the intact ear control (IEC). Under drought stress, ear photosynthesis was reduced by 46 % and 74.2 % at 25 DAA after removing the glume and awn, respectively. Under normal conditions, the number of ¹⁴C assimilates of DGC, and DAC was reduced by 14.6 % and 20.9 % in grains at 25 DAA, respectively, compared with the IEC. Compared with IED, the ¹⁴C assimilates of DGD, and DAD declined by 17.2 % and 27 %, respectively, in grains at 25 DAA under drought conditions. Under well-watered conditions, the grain weight per pot was reduced by 11.2 % and 25.4 % in the de-glumed control (DGC) and de-awned control (DAC) treatments, respectively, compared with the intact ear control (IEC). The grain weight per pot was further reduced after removing the glume and awn (16 % and 32.2 %, respectively) under drought stress. Furthermore, the awn contribution to grain weight was twice that of the glume. Our results suggest that the glume and awn of ears play prominent roles during grain filling in wheat, especially under drought stress, and that the awn is more crucial than the glume.

Low-temperature stress limits the growth and development of foxtail millet. Freezing stress caused by sudden temperature drops, such as late-spring coldness, often occurs in the seedling stage of foxtail millet. However, the ability and coping strategies of foxtail millet to cope with such stress are not clear. In the present study, we analyzed the self-regulatory mechanisms of freezing stress in foxtail millet. We conducted a physiological study on foxtail millet leaves at −4 °C for seven different durations (0, 2, 4, 6, 8, 10, and 12 h). Longer freezing time increased cell-membrane damage, relative conductance, and malondialdehyde content. This led to osmotic stress in the leaves, which triggered an increase in free proline, soluble sugar, and soluble protein contents. The increases in these substances helped to reduce the damage caused by stress. The activities of superoxide dismutase, peroxidase, and catalase increased reactive oxygen species (ROS) content. The optimal time point for the response to freezing stress was 8 h after exposure. The transcriptome analysis of samples held for 8 h at −4 °C revealed 6862 differentially expressed genes (DEGs), among which the majority are implicated in various pathways, including the starch and sucrose metabolic pathways, antioxidant enzyme pathways, brassinolide (BR) signaling pathway, and transcription factors, according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. We investigated possible crosstalk between BR signals and other pathways and found that BR signaling molecules were induced in response to freezing stress. The beta-amylase (BAM) starch hydrolase signal was enhanced by the BR signal, resulting in the accelerated degradation of starch and the formation of sugars, which served as emerging ROS scavengers and osmoregulators to resist freezing stress. In conclusion, crosstalk between BR signal transduction, and both starch and sucrose metabolism under freezing stress provides a new perspective for improving freezing resistance in foxtail millet.

The cytochrome P450 monooxygenases (CYP450) are the largest enzyme family in plant metabolism and widely involved in the biosynthesis of primary and secondary metabolites. Foxtail millet (Setaria italica (L.) P. Beauv) can respond to abiotic stress through a highly complex polygene regulatory network, in which the SiCYP450 family is also involved. Although the CYP450 superfamily has been systematically studied in a few species, the research on the CYP450 superfamily in foxtail millet has not been completed. In this study, three hundred and thirty-one SiCYP450 genes were identified in the foxtail millet genome by bioinformatics methods, which were divided into four groups, including forty-six subgroups. One hundred and sixteen genes were distributed in thirty-three tandem duplicated gene clusters. Chromosome mapping showed that SiCYP450 was distributed on seven chromosomes. In the SiCYP450 family of foxtail millet, 20 conserved motifs were identified. Cis-acting elements in the promoter region of SiCYP450 genes showed that hormone response elements were found in all SiCYP450 genes. Of the three hundred and thirty-one SiCYP450 genes, nine genes were colinear with the Arabidopsis thaliana genes. Two hundred SiCYP450 genes were colinear with the Setaria viridis genes, including two hundred and forty-five gene duplication events. The expression profiles of SiCYP450 genes in different organs and developmental stages showed that SiCYP450 was preferentially expressed in specific tissues, and many tissue-specific genes were identified, such as SiCYP75B6, SiCYP96A7, SiCYP71A55, SiCYP71A61, and SiCYP71A62 in the root, SiCYP78A1 and SiCYP94D9 in leaves, and SiCYP78A6 in the ear. The RT-PCR data showed that SiCYP450 could respond to abiotic stresses, ABA, and herbicides in foxtail millet. Among them, the expression levels of SiCYP709B4, SiCYP71A11, SiCYP71A14, SiCYP78A1, SiCYP94C3, and SiCYP94C4 were significantly increased under the treatment of mesotrione, florasulam, nicosulfuron, fluroxypyr, and sethoxydim, indicating that the same gene might respond to multiple herbicides. The results of this study will help reveal the biological functions of the SiCYP450 family in development regulation and stress response and provide a basis for molecular breeding of foxtail millet.

Soil salinization can lead to reduced soil activity, damage to the plant root system, stunted crop growth and reduced yield. Foxtail millet, an important cereal crop, has high nutritional and economic value but is affected by salt stress during growth. During long-term evolution, foxtail millet has developed various regulatory mechanisms to cope with salt stress. Among them, the glutathione S-transferase (GST) gene family plays a key role in the response to salt stress. GSTs are superfamily enzymes encoded by multiple genes with multiple functions that increase plant resistance to abiotic stresses through antioxidant defence and detoxification processes. We identified the gene SiGSTU24, which was the most highly upregulated gene under salt stress among the foxtail millet GST gene family, via screening. We found that, compared with WT and atgstu24 plants, Arabidopsis overexpressing SiGSTU24 presented a greater germination rate and taller plant height after salt stress. Moreover, we found that SiGSTU24 reduced ROS accumulation and changed the activities of antioxidant defence system enzymes in Arabidopsis. SiGSTU24 increased the expression of the antioxidant enzyme-related genes ascorbate peroxidase (AtAPX), superoxide dismutase (AtSOD), peroxidase (AtPOD), and catalase (AtCAT) in Arabidopsis. RNA-Seq and qRT‒PCR verification revealed that SiGSTU24 enhanced salt tolerance and antioxidant capacity in Arabidopsis under salt stress by regulating antioxidant enzyme-related genes and transcription factors. These genes and transcription factors may help Arabidopsis adapt to salt stress environments through various biochemical pathways and regulatory networks. SiGSTU24 overexpression enhances salt stress tolerance in Arabidopsis. This research provides a foundation for the study of SiGSTU24 function and supplements studies on gene functions in the foxtail millet GST gene family.