Abiotic stress is an important factor affecting turf establishment and limiting the sustainability of the turf industry. To alleviate the effects of combined cold and drought stress in cold- and drought-prone regions, it is essential to select and introduce turfgrass germplasms that are suitable for these conditions for successful turf establishment. Thus, we evaluated the effects of combined extreme cold and drought stress on the morphological, plant leaf functional, photosynthetic, and physiological and biochemical traits of 16 wild annual bluegrass (Poa annua) germplasms. We found that there were significant differences (P < 0.05) among different provenances, combined cold and drought stress, and the main interaction factors. Combined cold and drought stress altered the morphological characteristics of the 16 germplasms to varying degrees. Furthermore, combined cold and drought stress significantly reduced the net photosynthetic rate (Pn), stomatal conductance (gs), transpiration rate (Tr), instantaneous water use efficiency (WUE), chlorophyll content, chlorophyll fluorescence parameters, accumulated intercellular CO₂ concentration (Ci), and relative electrical conductivity (REC) and malondialdehyde (MDA), proline (Pro), soluble protein (SP), soluble sugar (SS), superoxide anion (O₂^.-), hydrogen peroxide (H₂O₂), and hydroxyl radical (·OH) and other active oxygen, and increased the superoxide dismutase activity (SOD), peroxidase activity (POD), catalase activity (CAT), ascorbate peroxidase activity (APX) and glutathione reductase (GR) activities. Comprehensive evaluation using principal component analysis (PCA), membership function analysis, and clustered heatmaps indicated that the ‘HZ’ germplasm had stronger combined cold and drought tolerance, whereas the ‘ZQ’ germplasm was more sensitive to combined cold and drought, which was roughly consistent with the order of morphological damage symptoms. Therefore, it is recommended to use the ‘HZ’ germplasm for planting projects in cold- and drought-prone areas, while the ‘ZQ’ germplasm is more suitable for use under warmer and non-water-deficient conditions.

Climate change and overgrazing significantly constrain the sustainability of meadow land and vegetation in the livestock industry on the Tibetan–Plateau ecosystem. In context of climate change mitigation, grassland soil C sequestration and forage sustainability, it is important to understand how manure regimes influence SOC stability, grassland soil, forage structure and nutritional quality. However, the responses of SOC fractions, soil and forage structure and quality to the influence of manure gradient practices remain unclear, particularly at Tianzhu belt, and require further investigation. A field study was undertaken to evaluate the soil bulk density, aggregate fractions and dynamics in SOC concentration, permanganate oxidizable SOC fractions, SOC stabilization and soil nutrients at the soil aggregate level under manure gradient practices. Moreover, the forage biodiversity, aboveground biomass and nutritional quality of alpine meadow plant communities were also explored. Four treatments, i.e., control (CK), sole sheep manure (SM), cow dung alone (CD) and a mixture of sheep manure and cow dung (SMCD) under five input rates, i.e., 0.54, 1.08, 1.62, 2.16 and 2.70 kg m⁻², were employed under randomized complete block design with four replications. Our analysis confirmed the maximum soil bulk density (BD) (0.80 ± 0.05 g cm⁻³) and micro-aggregate fraction (45.27 ± 0.77%) under CK, whilst the maximum macro-aggregate fraction (40.12 ± 0.54%) was documented under 2.70 kg m⁻² of SMCD. The SOC, very-labile C fraction (Cfrac₁), labile C fraction (Cfrac₂) and non-labile/recalcitrant C fraction (Cfrac₄) increased with manure input levels, being the highest in 2.16 kg m⁻² and 2.70 kg m⁻² applications of sole SM and the integration of 50% SM and 50% CD (SMCD), whereas the less-labile fraction (Cfrac₃) was highest under CK across aggregate fractions. However, manures under varying gradients improved SOC pools and stabilization for both macro- and micro-aggregates. A negative response of the carbon management index (CMI) in macro-aggregates was observed, whilst CMI in the micro-aggregate fraction depicted a positive response to manure addition with input rates, being the maximum under sole SM addition averaged across gradients. Higher SOC pools and CMI under the SM, CD and SMCD might be owing to the higher level of soil organic matter inputs under higher doses of manures. Moreover, the highest accumulation of soil nutrients,, for instance, TN, AN, TP, AP, TK, AK, DTPA extractable Zn, Cu, Fe and Mn, was recorded in SM, CD and SMCD under varying gradients over CK at both aggregate fractions. More nutrient accumulation was found in macro-aggregates over micro-aggregates, which might be credited to the physical protection of macro-aggregates. Overall, manure addition under varying input rates improved the plant community structure and enhanced meadow yield, plant community diversity and nutritional quality more than CK. Therefore, alpine meadows should be managed sustainably via the adoption of sole SM practice under a 2.16 kg m⁻² input rate for the ecological utilization of the meadow ecosystem. The results of this study deliver an innovative perspective in understanding the response of alpine meadows’ SOC pools, SOC stabilization and nutrients at the aggregate level, as well as vegetation structure, productivity and forage nutritional quality to manure input rate practices. Moreover, this research offers valuable information for ensuring climate change mitigation and the clean production of alpine meadows in the Qinghai–Tibetan Plateau area of China.

Rhizome is vital for carbon and nitrogen metabolism of the whole plant. However, the effect of carbon and nitrogen in the rhizome on rhizome expansion remains unclear. Three wild Kentucky bluegrass (Poa pratensis L.) germplasms with different rhizome expansion capacity (strong expansion capacity, ‘YZ’; medium expansion capacity, ‘WY’; and weak expansion capacity, ‘AD’) were planted in the field and the rhizomes number, tiller number, rhizome dry weight, physiological indicators and enzyme activity associated carbon and nitrogen metabolisms were measured. Liquid chromatography coupled to mass spectrometry (LC-MS) was utilized to analyze the metabolomic of the rhizomes. The results showed that the rhizome and tiller numbers of the YZ were 3.26 and 2.69-fold of that of the AD, respectively. The aboveground dry weight of the YZ was the greatest among all three germplasms. Contents of soluble sugar, starch, sucrose, NO₃⁻-N, and free amino acid were significantly higher in rhizomes of the YZ than those of the WY and AD (P < 0.05). The activities of glutamine synthetase (GS), glutamate dehydrogenase (GDH) and sucrose phosphate synthase (SPS) of the YZ were the highest among all three germplasm, with values of 17.73 A·g^− 1 h^− 1, 5.96 µmol·g^− 1 min^− 1, and 11.35 mg·g^− 1 h^− 1, respectively. Metabolomics analyses revealed that a total of 28 differentially expressed metabolites (DEMs) were up-regulated, and 25 DEMs were down-regulated in both comparison groups (AD vs. YZ group and WY vs. YZ group). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that metabolites related to histidine metabolism, tyrosine metabolism, tryptophan metabolism, and phenylalanine metabolism were associated with rhizomes carbon and nitrogen metabolism. Overall, the results suggest that soluble sugar, starch, sucrose, NO₃⁻-N, and free amino acid in rhizome are important to and promote rhizome expansion in Kentucky bluegrass, while tryptamine, 3-methylhistidine, 3-indoleacetonitrile, indole, and histamine may be key metabolites in promoting carbon and nitrogen metabolism of rhizome.

Combined drought and low–temperature stress is a crucial factor affecting turfgrass establishment and limiting the sustainability of the turfgrass industry in drought– and cold–prone regions. In this context, we evaluated the effects of regular watering (the soil water content was 80% of the maximum water–holding capacity of the field) at room temperature (25 °C) and combined drought (the soil water content was 30% of the maximum water–holding capacity of the field) and low–temperature (0 °C) stress on the morphology, photosynthesis, and physiology of wild Poa annua seedlings from different provenances (‘PA’, ‘WY’, ‘NX’ and ‘YC’). Results indicated that the combined drought and low–temperature stress changed the morphological and growth indicators of seedlings in four provenances to different extents. Moreover, combined drought and low–temperature stress reduced the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), water use efficiency (WUE), and chlorophyll content in seedlings from four provenances. However, intertemporal CO₂ concentration (Ci), relative electrical conductivity (REC), the contents of malondialdehyde (MDA), proline (Pro), soluble sugars (SS), the superoxide anion (O₂^•−) production rate, the contents of hydrogen peroxide (H₂O₂) and hydroxyl radical (·OH), the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were all increased. The increase in ‘PA’ was much greater than that in ‘NX.’ The comprehensive evaluation results showed that the order of combined drought and low–temperature resistance of seedlings from the four provenances was ‘PA’ > ‘YC’ > ‘WY’ > ‘NX’, which corresponded to the order of the morphological damage symptoms. In conclusion, ‘PA’ may maintain stronger combined drought and low–temperature resistance by improving the cellular water absorption and retention capacity, enhancing the function of the antioxidant defense system, and maintaining the integrity of the cell membrane, which is a crucial germplasm resource for breeding combined drought and low–temperature resistance in Poa annua.

Rhizome growth and development is regulated by phytohormone. However, endogenous phytohormones affect rhizome initiation, and sustained growth in perennial grass species remains elusive. In this study, we investigated the morphological characteristics and the content of indole-3-acetic acid (IAA), zeatin (ZT), gibberellic acid (GA3), and abscisic acid (ABA) in the rhizomes of two different Kentucky bluegrass. Using ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS), we performed metabolite analysis of two different rhizomes. In our study, the multi-rhizome Kentucky bluegrass material ‘Yuzhong’ had an average of 1113 rhizomes, while the few-rhizome material ‘Anding’ had an average of 347 rhizomes. The diameter of rhizome and length of rhizome internode in ‘Yuzhong’ were 1.68-fold and 1.33-fold higher than that of the ‘Anding’, respectively. The rhizome dry weight of ‘Yuzhong’ was 75.06 g, while the ‘Anding’ was 20.79 g. ‘Yuzhong’ had a higher ZT content (5.50 μg·g⁻¹), which is 2.4-fold that of ‘Anding’ (2.27 μg·g⁻¹). In contrast, the IAA, ABA, and GA3 content of rhizome were markedly higher in ‘Anding’ than ‘Yuzhong’. Correlation analysis revealed significant correlations between ZT and ZT/ABA with the number of rhizomes, diameter of rhizome, and length of rhizome internode, whereas IAA, ABA, GA3, and IAA/ZT were opposite. In the metabolic profiles, we identified 163 differentially expressed metabolites (DEMs) (60 upregulated and 103 downregulated) in positive ion mode and 75 DEMs (36 upregulated and 39 downregulated) in negative ion mode. Histidine metabolism and ABC transporters pathways were the most significantly enriched in the positive and negative ion mode, respectively, both of which are involved in the synthesis and transport of cytokinin. These results indicate that cytokinin is crucial for rhizome development and promotes rhizome germination and growth of Kentucky bluegrass.