Drought is one of the most important factors affecting plant growth and production due to ongoing global climate change. Elymus sibiricus has been widely applied for ecological restoration and reseeding of degraded grassland in the Qinghai–Tibetan Plateau (QTP) because of its strong adaptability to barren, salted, and drought soils. To explore the mechanism of drought resistance in E. sibiricus, drought-tolerant and drought-sensitive genotypes of E. sibiricus were used in metabolomic studies under simulated long-term and short-term drought stress. A total of 1091 metabolites were detected, among which, 27 DMs were considered to be the key metabolites for drought resistance of E. sibiricus in weighted gene co-expression network analysis (WGCNA). Ten metabolites, including 3-amino-2-methylpropanoic acid, coniferin, R-aminobutyrate, and so on, and 12 metabolites, including L-Proline, L-histidine, N-acetylglycine, and so on, showed differential accumulation patterns under short-term and long-term drought stress, respectively, and thus, could be used as biomarkers for drought-tolerant and drought-sensitive E. sibiricus. In addition, different metabolic accumulation patterns and different drought response mechanisms were also found in drought-tolerant and drought-sensitive genotypes of E. sibiricus. Finally, we constructed metabolic pathways and metabolic patterns for the two genotypes. This metabolomic study on the drought stress response of E. sibiricus can provide resources and a reference for the breeding of new drought-tolerant cultivars of E. sibiricus.

Galega orientalis, a leguminous herb in the Fabaceae family, is an ecologically and economically important species widely cultivated for its strong stress resistance and high protein content. However, genomic information of Galega orientalis has not been reported, which limiting its evolutionary analysis. The small genome size makes chloroplast relatively easy to obtain genomic sequence for phylogenetic studies and molecular marker development. Here, the chloroplast genome of Galega orientalis was sequenced and annotated. The results showed that the chloroplast genome of G. orientalis is 125,280 bp in length with GC content of 34.11%. A total of 107 genes were identified, including 74 protein-coding genes, 29 tRNAs and four rRNAs. One inverted repeat (IR) region was lost in the chloroplast genome of G. orientalis. In addition, five genes (rpl22, ycf2, rps16, trnE-UUC and pbf1) were lost compared with the chloroplast genome of its related species G. officinalis. A total of 84 long repeats and 68 simple sequence repeats were detected, which could be used as potential markers in the genetic studies of G. orientalis and related species. We found that the Ka/Ks values of three genes petL, rpl20, and ycf4 were higher than one in the pairwise comparation of G. officinalis and other three Galegeae species (Calophaca sinica, Caragana jubata, Caragana korshinskii), which indicated those three genes were under positive selection. A comparative genomic analysis of 15 Galegeae species showed that most conserved non-coding sequence regions and two genic regions (ycf1 and clpP) were highly divergent, which could be used as DNA barcodes for rapid and accurate species identification. Phylogenetic trees constructed based on the ycf1 and clpP genes confirmed the evolutionary relationships among Galegeae species. In addition, among the 15 Galegeae species analyzed, Galega orientalis had a unique 30-bp intron in the ycf1 gene and Tibetia liangshanensis lacked two introns in the clpP gene, which is contrary to existing conclusion that only Glycyrrhiza species in the IR lacking clade (IRLC) lack two introns. In conclusion, for the first time, the complete chloroplast genome of G. orientalis was determined and annotated, which could provide insights into the unsolved evolutionary relationships within the genus Galegeae.

Soil salinization is a significant obstacle to improving natural grasslands, also affecting farmland productivity. The main distribution area of high salinity soils is primarily located in western China, with a significant presence in the Qinghai–Tibet Plateau (QTP). Siberian wildrye (Elymus sibiricus L.) has been widely used in ecological restoration and reseeding in the QTP, due to its strong salt and alkali resistance. To comprehensively evaluate its salt tolerance and to screen salt-tolerant and salt-sensitive germplasm resources, 50 wild E. sibiricus accessions were assessed through the measurement of several physiological parameters. These included the relative water content (RWC); relative electrical conductivity (REC); malondialdehyde content (MDA); proline content (Pro); and Na⁺/K⁺ ratio after 14 days of salt stress treatment (200 mmol L⁻¹ NaCl). Through principal component analysis (PCA) based on both material sources and physiological indexing, it was found that the salt damage score (SDS) and RWC had a strong positive correlation (p < 0.05), in opposition to other indices. The positive indicators were REC, Pro, Na⁺/K⁺, and MDA, and the negative indicators were RWC and SDS. Ultimately, by combining the results of the conventional membership function analysis and the weighted membership function analysis based on major components, three salt-tolerant accessions—including XJ013-28, NM021-13 and XJ012-17—and four salt-sensitive accessions, including XZ008-20, QH013-8, XZ009-20, and QH005-35, were screened out. These results could be effectively used to assess the effects of salt stress on Siberian wildrye and its related species. This study provides a reference for the discovery and breeding of salt-tolerant cultivars among the Elymus species.

Genetic markers play a central role in understanding genetic diversity, speciation, evolutionary processes, and how species respond to environmental stresses. However, conventional molecular markers are less effective when studying polyploid species with large genomes. In this study, we compared gene expression levels in 101 accessions of Elymus sibiricus, a widely distributed allotetraploid forage species across the Eurasian continent. A total of 20,273 high quality transcriptomic SNPs were identified. In addition, 72,344 evolutionary information loci of these accessions of E. sibiricus were identified using genome skimming data in conjunction with the assembled composite genome. The population structure results suggest that transcriptome SNPs were more effective than SNPs derived from genome skimming data in revealing the population structure of E. sibiricus from different locations, and also outperformed gene expression levels. Compared with transcriptome SNPs, the investigation of population-specifically-expressed genes (PSEGs) using expression levels revealed a larger number of locally adapted genes mainly involved in the ion response process in the Sichuan, Inner Mongolia, and Xizang geographical groups. Furthermore, we performed the weighted gene co-expression network analysis (WGCNA) and successfully identified potential regulators of PSEGs. Therefore, for species lacking genomic information, the use of transcriptome SNPs is an efficient approach to perform population structure analysis. In addition, analyzing genes under selection through nucleotide diversity and genetic differentiation index analysis based on transcriptome SNPs, and exploring PSEG through expression levels is an effective method for analyzing locally adaptive genes.

Kengyilia is a newly established genus. Most species in this genus survive in hash environment, which might be an indicator of an acquirement of stress resistance genes and the potential for molecular breeding in Triticeae species. Quantitative real-time PCR (qRT-PCR) is a widely used technique with varied sensitivity heavily dependent on the optimal level of the reference genes. K. melanthera is a typical psammophyte species which has high drought resistance. The reference genes of K. melanthera are not yet reported. This study aims to evaluate the expression stability of 14 candidate reference genes (EF1A, GAPDH, ACT1, UBI, TUBB3, TIPRL, CACS, PPP2R1B, TUBA1A, EIF4A1, CYPA3, TCTP, ABCG11L, and FBXO6L) under five treatments (drought, heat, cold, salt, and ABA) and find the most stable and suitable one even upon stressed conditions. The software NormFinder, GeNorm, BestKeeper, and RefFinder were used for data analysis. In general, the genes CACS and PPP2R1B are concluded to have the best overall performance under the various treatments. With the ABA treatment, TCTP and TIPRL show the best stability. CACS and TCTP, as well as TIPRL and CYPA3, were most stable under the treatments of cold and salt, respectively. CACS and FBXO6L were ranked the highest with the heat treatment and drought treatment, respectively. Finally, the Catalase-1 (CAT1) gene was used to verify the reliability of the above reference genes. Accordingly, CAT1’s expression pattern remained unchanged after normalization with stable reference genes. This study provides beneficial information about the stability and reliability of potential reference genes for qRT-PCR in K. melanthera.

Orchardgrass (Dactylis glomerata L.) is highly tolerant of shade, cold, and overwintering, making it an ideal species for grassland ecological restoration and livestock production. However, the genetic diversity of orchardgrass may be threatened by climate change. Using a Maximum Entropy (MaxEnt) model with the BCC-CSM2-MR global climate database and the Harmonized World Soil Database, we projected the current and future distribution of orchardgrass suitable areas globally. The predicted ecological thresholds for vital environmental factors were determined to be a temperature seasonality range of 411.50–1034.37 °C, a mean diurnal range of −0.88–10.69 °C, a maximum temperature of the warmest month of 22.21–35.45 °C, and precipitation of the coldest quarter of 116.56–825.40 mm. A range of AUC values from 0.914 to 0.922, indicating the accuracy of the prediction model. Our results indicate that the total area of current suitable habitats for orchardgrass was estimated to be 2133.01 × 10⁴ km², it is dispersed unevenly over six continents. Additionally, the suitable areas of habitats increased in higher latitudes while decreasing in lower latitudes as greenhouse gas emissions increased. Therefore, efforts should be made to save places in the southern hemisphere that are in danger of becoming unsuitable, with the possibility of using northern America, China, and Europe in the future for conservation and extensive farming.