Seed aging has an important effect on the germplasm preservation and industrialized production of Siberian wildrye (Elymus sibiricus) in the Qinghai-Tibet Plateau. However, so far its underlying molecular mechanisms still largely remain unknown. To shed light on this topic, one-year stored seeds of E. sibiricus were exposed to artificial aging treatments (AAT), followed by seed vigor characteristics and physiological status monitoring. Then global proteomics analysis was undertaken by the tandem mass tags (TMT) technique, and the proteins were quantified with liquid chromatography-tandem mass spectrometry on three aging time points (0 h, 36 h and 72 h). Finally, we verified the expression of related proteins by parallel reaction monitoring (PRM). Our results demonstrated that the seed vigor decreased remarkably in response to artificial aging, but the relative ion-leakage and malondialdehyde content, superoxide anion and hydrogen peroxide showed the opposite situation. Proteomic results showed that a total of 4169 proteins were identified and quantified. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that a series of key pathways including carbohydrate metabolism, lipid metabolism, and antioxidant activity were severely damaged by aging treatments. Numerous key proteins such as glyceraldehyde triphosphate glyceraldehyde dehydrogenase, succinate dehydrogenase, lipoxygenase, peroxidase, glutathione-s-transferase and late embryogenesis abundant proteins were significantly down-regulated. However, the up-regulation of the heat shock protein family has made a positive contribution to oxidative stress resistance in seeds. This study provides a useful catalog of the E. sibiricus proteomes with insights into the future genetic improvement of seed storability.

Oats (Avena sativa) are an important cereal crop and cool-season forage worldwide. Heat shock protein 90 (HSP90) is a protein ubiquitously expressed in response to heat stress in almost all plants. To date, the HSP90 gene family has not been comprehensively reported in oats. Herein, we have identified twenty HSP90 genes in oats and elucidated their evolutionary pathways and responses to five abiotic stresses. The gene structure and motif analyses demonstrated consistency across the phylogenetic tree branches, and the groups exhibited relative structural conservation. Additionally, we identified ten pairs of segmentally duplicated genes in oats. Interspecies synteny analysis and orthologous gene identification indicated that oats share a significant number of orthologous genes with their ancestral species; this implies that the expansion of the oat HSP90 gene family may have occurred through oat polyploidization and large fragment duplication. The analysis of cis-acting elements revealed their influential role in the expression pattern of HSP90 genes under abiotic stresses. Analysis of oat gene expression under high-temperature, salt, cadmium (Cd), polyethylene glycol (PEG), and abscisic acid (ABA) stresses demonstrated that most AsHSP90 genes were significantly up-regulated by heat stress, particularly AsHSP90-7, AsHSP90-8, and AsHSP90-9. This study offers new insights into the amplification and evolutionary processes of the AsHSP90 protein, as well as its potential role in response to abiotic stresses. Furthermore, it lays the groundwork for understanding oat adaptation to abiotic stress, contributing to research and applications in plant breeding.

This study aimed to investigate the effect of oat silage treated with a low-temperature-tolerant lactic acid bacteria (LAB) inoculant on milk yield and the quality of lactating yaks. Oat silages were prepared in big round bales, treated without (control) or with a low-temperature-tolerant LAB inoculant (a mixture of Lactobacillus plantarum BP18, Pediococcus pentosaceus HS1 and Lactobacillus buchneri LP22; the application rate of 10⁵ cfu/g on a fresh matter basis). Eighteen lactating yaks were divided into nine pairs with a similar milk yield. Each pair of yaks was randomly allocated to the control or LAB-inoculated silage treatment. The inoculated silage increased the dry matter intake and the total volatile fatty acid (mainly acetate, propionate and butyrate) in rumen fluid compared with the control. The inoculated silage also enhanced the yield of yak milk with high contents of total N, fat and lactose. In addition, high levels of essential amino acids (Thr, Leu and Phe), polyunsaturated fatty acids and low saturated fatty acids were observed in milk when lactating yaks were fed with the inoculated silage. Therefore, inoculation with a low-temperature-tolerant LAB during ensiling could promote the milk yield of lactating yaks by enhancing dry matter intake and ruminal fermentation.

The objective of this experiment was to determine whether compound microbial inoculants could enhance the fermentation of oat and common vetch silage that were stored in the Northwest Sichuan Plateau for 60 days under extremely low temperatures. Oat and common vetch harvested from single and mixed artificially planted grassland of oat and common vetch were chopped into 2–3 cm (oat, S1; common vetch, S2; oat–common vetch = 2:1, S3), then sterile water (T1), Zhuang Lemei IV silage additive (T2), and Fu Zhengxing silage additive (T3) were added to the feed and ensiled at the local outdoor environment for 60 days. Data were analyzed as a 3 × 3 factorial arrangement of treatments with the main effects of the materials, additives, and their interaction. Interactions between the materials and additives significantly affected the fermentation quality and the content of DM, WSC, and NDF and the number of yeasts in forages. Treatments with S3 have significantly higher contents of lactic acid, acetic acid, and lactic acid bacteria in the feed than those in the S1 and S2 treatments, while the contents of AN/TN and propionic acid were significantly lower compared with the S1 and S2 treatments (p < 0.05). Concentrations of lactic acid, acetic acid, and propionic acid were significantly increased and the content of neutral detergent fiber in the T2-treated silage decreased compared with the T1 treatment (p < 0.05). The T3 treatment significantly reduced the number of yeasts in the silage but the compound lactic acid bacteria additive treatment (T1, T2) significantly decreased the butyric acid content and pH of the feed and increased the acid detergent fiber content and the number of lactic acid bacteria in the feed compared with the T1 treatment. Among them, the butyric acid content of the T3 treatment decreased by 63.64–86.05%, while that of the T2 treatment decreased by 36.36–83.33% (p < 0.05). The comprehensive analysis of the membership function revealed that the silage quality was the best after the S3T2 treatment, so the implementation of the S3T2 combination in the Northwest Sichuan Plateau can provide guarantees for the production of local high-quality forage grass and alleviate the shortage of forage grass.

Reed canary grass (Phalaris arundinacea L.) is known for its tolerance to drought, heavy metals, and waterlogging, making it a popular choice for forage production and wetland restoration in the Qinghai-Tibet Plateau (QTP). To accurately assess gene expression in reed canary grass under different abiotic stresses, suitable reference genes need to be identified and validated. Thirteen candidate reference gene sequences were selected and screened using RT-qPCR to detect their expression levels in reed canary grass leaves under drought, salt, cadmium, and waterlogging stresses. Four algorithms were used to assess the stability of the expression levels of the candidate reference genes. The most stably expressed genes were UBC and H3 under drought Cd, ETF and CYT under salt stress, and ETF and TUB under waterlogging stress. GAPDH was found to be less stable under abiotic stresses. PIP-1, PAL, NAC 90, and WRKY 72A were selected as response genes for quantitative expression assessment under drought, salt, Cd, and waterlogging stresses to confirm the accuracy of the selected stable reference genes. These results provide a theoretical reference for assessing gene expression in reed canary grass under abiotic stresses.

Plants cultivated on the Qinghai-Tibet Plateau (QTP) are exposed to high ultraviolet radiation intensities, so they require effective mechanisms to adapt to these stress conditions. UV-B radiation is an abiotic stress factor that affects plant growth, development, and environmental adaptation. Elymus sibiricus is a common species in the alpine meadows of the QTP, with high-stress resistance, large biomass, and high nutritional value. This species plays an important role in establishing artificial grasslands and improving degraded grasslands. In this study, UV-B radiation-tolerant and UV-B radiation-sensitive E. sibiricus genotypes were subjected to simulated short-term (5 days, 10 days) and long-term (15 days, 20 days) UV-B radiation stress and the metabolite profiles evaluated to explore the mechanism underlying UV-B radiation resistance in E. sibiricus. A total of 699 metabolites were identified, including 11 primary metabolites such as lipids and lipid-like molecules, phenylpropanoids and polyketides, organic acids and their derivatives, and organic oxygen compounds. Principal component analysis distinctly clustered the samples according to the cultivar, indicating that the two genotypes exhibit distinct response mechanisms to UV-B radiation stress. The results showed that 14 metabolites, including linoleic acid, LPC 18:2, xanthosine, and 23 metabolites, including 2-one heptamethoxyflavone, glycyrrhizin, and caffeic acid were differentially expressed under short-term and long-term UV-B radiation stress, respectively. Therefore, these compounds are potential biomarkers for evaluating E. sibiricus response to UV-B radiation stress. Allantoin specific and consistent expression was up-regulated in the UV-B radiation-tolerant genotype, thereby it can be used to identify varieties resistant to UV-B radiation. Different metabolic profiles and UV-B radiation response mechanisms were observed between the UV-B radiation-tolerant and UV-B radiation-sensitive E. sibiricus genotypes. A model for the metabolic pathways and metabolic profiles was constructed for the two genotypes. This metabolomic study on the E. sibiricus response to UV-B radiation stress provides a reference for the breeding of new UV-B radiation-tolerant E. sibiricus cultivars.