Hydrotime Model Parameters Estimate Seed Vigor and Predict Seedling Emergence Performance of Astragalus sinicus under Various Environmental Conditions.

Seed vigor is an important aspect of seed quality. High-vigor seeds show rapid and uniform germination and emerge well, especially under adverse environmental conditions. Here, we determined hydrotime model parameters by incubating seeds at different water potentials (0.0, -0.2, -0.4, -0.6, and -0.8 MPa) in the laboratory, for 12 seed lots of Chinese milk vetch (Astragalus sinicus) (CMV), a globally important legume used as forage, green manure, and a rotation crop. Pot experiments were conducted to investigate the seedling emergence performance of 12 CMV seed lots under control, water stress, salinity stress, deep sowing, and cold stress conditions. Meanwhile, the field emergence performance was evaluated on two sowing dates in June and October 2022. Correlation and regression analyses were implemented to explore the relationships between hydrotime model parameters and seedling emergence performance under various environmental conditions. The seed germination percentage did not differ significantly between seed lots when seeds were incubated at 0.0 MPa, whereas it did differ significantly between seed lots at water potentials of -0.2, -0.4, and -0.6 MPa. The emergence percentage, seedling dry weight, and simplified vigor index also differed significantly between the 12 seed lots under various environmental conditions. Ψb(50) showed a significant correlation with germination and emergence performance under various environmental conditions; however, little correlation was observed between θH or σφb and germination and emergence. These results indicate that Ψb(50) can be used to estimate seed vigor and predict seedling emergence performance under diverse environmental conditions for CMV and similar forage legumes. This study will enable seed researchers, plant breeders, and government program directors to target higher seed vigor more effectively for forage legumes.

Microbial network structure, not plant and microbial community diversity, regulates multifunctionality under increased precipitation in a cold steppe.

It is known that the dynamics of multiple ecosystem functions (i. e., multifunctionality) are positively associated with microbial diversity and/or biodiversity. However, how the relationship between microbial species affects ecosystem multifunctionality remains unclear, especially in the case of changes in precipitation patterns. To explore the contribution of biodiversity and microbial co-occurrence networks to multifunctionality, we used rainfall shelters to simulate precipitation enhancement in a cold steppe in Northeast China over two consecutive growing seasons. We showed that an increased 50% precipitation profoundly reduced bacterial diversity and multidiversity, while inter-annual differences in precipitation did not shift microbial diversity, plant diversity, or multidiversity. Our analyses also revealed that increased annual precipitation significantly increased ecosystem, soil, nitrogen, and phosphorous cycle multifunctionality. Neither increased precipitation nor inter-annual differences in precipitation had a significant effect on carbon cycle multifunctionality, probably due to the relatively short period (2 years) of our experiment. The co-occurrence network of bacterial and fungal communities was the most dominant factor affecting multifunctionality, the numbers of negative interactions but not positive interactions were linked to multifunctionality. In particular, our results provided evidence that microbial network topological features are crucial for maintaining ecosystem functions in grassland ecosystems, which should be considered in related studies to accurately predict the responses of ecosystem multifunctionality to predicted changes in precipitation patterns.

Germination ecology of Chenopodium album L. and implications for weed management.

Chenopodium album L. is a troublesome annual species in various cropping systems, and a sound knowledge of the ecological response of C. album germination to environmental factors would suggest suitable management strategies for inhibiting its spread. Preliminary laboratory-based research was conducted to investigate germination and emergence requirements of C. album under various environmental conditions (e.g., photoperiods, constant temperature, salinity, moisture, soil pH, burial depth, and oat crop residue). Results showed C. album seeds were found to be photoblastic, with only 13% germination in darkness. The maximum germination (94%) of C. album occurred at an optimal temperature of 25°C, and the depressive effect of other temperatures on germination was more severe at lower rather than higher temperatures. Seed germination was suitably tolerant of salinity and osmotic potential, with germination observed at 200 mM NaCl (37.0%) and -0.8 MPa (20%), respectively. Germination was relatively uniform (88-92%) at pH levels ranging from 4 to 10. The maximum germination of C. album was observed on the soil surface, with no or rare emergence of seeds at a burial depth of 2 cm or under 7000 kg ha-1 oat straw cover, respectively. Information provided by this study will help to develop more sustainable and effective integrated weed management strategies for the control of C. album, including (i) a shallow-tillage procedures to bury weed seeds in conventional-tillage systems and (ii) oat residue retention or coverage on the soil surface in no-tillage systems.

Mowing enhances the positive effects of nitrogen addition on ecosystem carbon fluxes and water use efficiency in a semi-arid meadow steppe.

Grasslands are now facing a continuously increasing supply of nitrogen (N) fertilizers, resulting in alterations in ecosystem functioning, including changes in carbon (C) and water cycling. Mowing, one of the most widely used grassland management techniques, has been shown to mitigate the negative impacts of increased N availability on species richness. However, knowledge of how N addition and mowing, alone and/or in combination, affect ecosystem-level C fluxes and water use efficiency (WN) is still limited. We experimentally manipulated N fertilization (0 and 10 g N m-2 yr-1) and mowing (once per year at the end of the growing season) following a randomized block design in a meadow steppe characterized by salinization and alkalinization in northeastern China. We found that, compared to the control plots, N addition, mowing, and their interaction increased net ecosystem CO2 exchange by 65.1%, 14.7%, and 133%, and WN by 40.7%, 18.5%, and 96.1%, respectively. Nitrogen enrichment also decreased soil pH, which resulted in greater aboveground biomass (AGB). Moreover, N addition indirectly increased AGB by inducing changes in species richness. Our results indicate that mowing enhances the positive effects of N addition on ecosystem C fluxes and WN. Therefore, appropriate grassland management practices are essential to improve ecosystem C sequestration, WN, and mitigate future species diversity declines due to ecosystem eutrophication.

Effect of Light Intensity on Morphology, Photosynthesis and Carbon Metabolism of Alfalfa (Medicago sativa) Seedlings.

To understand how light intensity influences plant morphology and photosynthesis in the forage crop alfalfa (Medicago sativa L. cv. Zhongmu 1), we investigated changes in leaf angle orientation, chlorophyll fluorescence, parameters of photosynthesis and expression of genes related to enzymes involved in photosynthesis, the Calvin cycle and carbon metabolism in alfalfa seedlings exposed to five light intensities (100, 200, 300, 400 and 500 µmol m-2 s-1) under hydroponic conditions. Seedlings grown under low light intensities had significantly increased plant height, leaf hyponasty, specific leaf area, photosynthetic pigments, leaf nitrogen content and maximal PSII quantum yield, but the increased light-capturing capacity generated a carbon resource cost (e.g., decreased carbohydrates and biomass accumulation). Increased light intensity significantly improved leaf orientation toward the sun and upregulated the genes for Calvin cycle enzymes, thereby increasing photosynthetic capacity. Furthermore, high light (400 and 500 µmol m-2 s-1) significantly enhanced carbohydrate accumulation, accompanied by gene upregulation and increased activity of sucrose and starch-synthesis-related enzymes and those involved in carbon metabolism. Together, these results advance our understanding of morphological and physiological regulation in shade avoidance in alfalfa, which would guide the identification of suitable spatial planting patterns in the agricultural system.

Integrative analysis of the metabolome and transcriptome reveal the phosphate deficiency response pathways of alfalfa.

Understanding the mechanisms underlying the responses to inorganic phosphate (Pi) deficiency in alfalfa will help enhance Pi acquisition efficiency and the sustainable use of phosphorous resources. Integrated global metabolomic and transcriptomic analyses of mid-vegetative alfalfa seedlings under 12-day Pi deficiency were conducted. Limited seedling growth were found, including 13.24%, 16.85% and 33.36% decreases in height, root length and photosynthesis, and a 24.10% increase in root-to-shoot ratio on day 12. A total of 322 and 448 differentially abundant metabolites and 1199 and 1061 differentially expressed genes were identified in roots and shoots. Increased (>3.68-fold) inorganic phosphate transporter 1;4 and SPX proteins levels in the roots (>2.15-fold) and shoots (>2.50-fold) were related to Pi absorption and translocation. The levels of phospholipids and Pi-binding carbohydrates and nucleosides were decreased, while those of phosphatases and pyrophosphatases in whole seedlings were induced under reduced Pi. In addition, nitrogen assimilation was affected by inhibiting high-affinity nitrate transporters (NRT2.1 and NRT3.1), and nitrate reductase. Increased delphinidin-3-glucoside might contribute to the gray-green leaves induced by Pi limitation. Stress-induced MYB, WRKY and ERF transcription factors were identified. The responses of alfalfa to Pi deficiency were summarized as local systemic signaling pathways, including root growth, stress-related responses consisting of enzymatic and nonenzymatic systems, and hormone signaling and systemic signaling pathways including Pi recycling and Pi sensing in the whole plant, as well as Pi recovery, and nitrate and metal absorption in the roots. This study provides important information on the molecular mechanism of the response to Pi deficiency in alfalfa.

Nitrogen addition amplifies the nonlinear drought response of grassland productivity to extended growing-season droughts.

Understanding the response of grassland production and carbon exchange to intra-annual variation in precipitation and nitrogen addition is critical for sustainable grassland management and ecosystem restoration. We introduced growing-season drought treatments of different lengths (15, 30, 45 and 60 d drought) by delaying growing-season precipitation in a long-term nitrogen addition experiment in a low diversity meadow steppe in northeast China. Response variables included aboveground biomass (AGB), ecosystem net carbon exchange (NEE), and leaf net carbon assimilation rate (A). In unfertilized plots drought decreased AGB by 13.7% after a 45-d drought and 31.7% after a 60-d drought (47.6% in fertilized plots). Progressive increases in the drought response of NEE were also observed. The effects of N addition on the drought response of productivity increased as drought duration increased, and these responses were a function of changes in AGB and biomass allocation, particularly root to shoot ratio. However, no significant effects of drought occurred in fertilized or unfertilized plots in the growing season a year after the experiment, N addition did limit the recovery of AGB from severe drought during the remainder of the current growing season. Our results imply that chronic N enrichment could exacerbate the effects of growing-season drought on grassland productivity caused by altered precipitation seasonality under climate change, but that these effects do not carry over to the next growing season.

Towards a mechanistic understanding of soil nitrogen availability responses to summer vs. winter drought in a semiarid grassland.

More frequent and intense drought events resulting from climate change are anticipated to become important drivers of change for terrestrial ecosystem function by affecting water and nutrient cycles. In semiarid grasslands, the responses of soil nitrogen availability to severe drought and the underlying mechanisms are largely unknown. Moreover, the responses and mechanisms may vary between summer and winter drought. We examined soil nitrogen availability responses to extreme reductions in precipitation over summer and winter using a field experiment in a semiarid grassland located in northeast China, and we explored the mechanisms by examining associated changes in abiotic factors (soil property responses) and biotic factors (plant and soil microbial responses). The results demonstrated that both the summer and winter severe drought treatments significantly reduced plant and microbial biomass, whereas summer drought also changed soil microbial community structure. Summer drought, winter drought and combined summer and winter drought decreased the resistance of soil nitrogen availability by 38.7 ± 11.1%, 43.3 ± 11.4% and 43.8 ± 6.0%, respectively. While both changes in abiotic factors (reduced soil water content and total nitrogen content) and biotic factors (reduced plant and microbial biomass) explained the resistance of soil nitrogen availability to drought over summer, only changes in biotic factors (reduced plant and microbial biomass) explained the legacy effect of winter drought. Our results highlight that severe drought can have important consequences for nitrogen cycling in semiarid grasslands, and that both the effects of summer and winter drought must be accounted for in predicting these responses.

Drought sensitivity of aboveground productivity in Leymus chinensis meadow steppe depends on drought timing.

There is limited understanding of the combined effects of discrete climate extremes and chronic environmental changes on ecosystem processes and functioning. We assessed the interactions of extreme drought timing (45 days, in spring or summer) and nitrogen (N) addition in a full factorial field experiment in a Leymus chinensis-dominated meadow steppe in northeast China. We evaluated the resistance and recovery of the grassland (calculated in terms of aboveground biomass) to these two drought events. The spring drought reduced aboveground biomass by 28% in the unfertilized plots and by 33% in the fertilized plots, and the effects persisted during the subsequent post-drought period within the same growing season; however, the summer drought had no significant influence on aboveground biomass. Although there were no significant interactive effects between drought timing and N addition, we observed a potential trend of N addition increasing the proportion of aboveground biomass suppressed by spring drought but not summer drought. Moreover, the drought resistance of the aboveground biomass was positively correlated with the response of the belowground biomass to drought. One year after the extreme drought events, the spring drought effects on aboveground and belowground biomass were negligible. Our results indicate that the drought sensitivity of productivity likely depends on the phenological and morphological traits of the single highly dominant species (Leymus chinensis) in this meadow steppe.

Nitrogen Addition Increases the Sensitivity of Photosynthesis to Drought and Re-watering Differentially in C3 Versus C4 Grass Species.

Global change factors, such as variation in precipitation regimes and nitrogen (N) deposition, are likely to occur simultaneously and may have profound impacts on the relative abundance of grasses differing in functional traits, such as C3 and C4 species. We conducted an extreme drought and re-watering experiment to understand differences in the resistance and recovery abilities of C3 and C4 grasses under different N deposition scenarios. A C3 perennial grass (Leymus chinensis) and two C4 grasses (annual species Chloris virgata and perennial species Hemarthria altissima) that co-occur in Northeast China were selected as experimental plants. For both C3 and C4 grasses, N addition caused a strong increase in biomass and resulted in more severe drought stress, leading to a change in the dominant photosynthetic limitation during the drought periods. Although N addition increased antioxidant enzyme activities and protective solute concentrations, the carbon fixing capacity did not fully recover to pre-drought levels by the end of the re-watering period. N addition resulted in lower resilience under the drought conditions and lower resistance at the end of the re-watering. However, N addition led to faster recovery of photosynthesis, especially in the C3 grass, which indicate that the effect of N addition on photosynthesis during drought was asymmetric, especially in the plants with different photosynthetic nitrogen use efficiency (PNUE). These findings demonstrated that nitrogen deposition may significant alter the susceptibility of C3 and C4 grass species to drought stress and re-watering, highlighting the asymmetry between resistance and resilience and to improve our understanding about plant responses to climate change.

Effects of Nitrogen Addition on the Drought Susceptibility of the Leymus chinensis Meadow Ecosystem Vary with Drought Duration.

It is not clear yet how extreme drought and nitrogen (N) deposition influence grassland ecosystem functions when they are considered together, especially in complex field conditions. To explore the response of the Leymus chinensis meadow ecosystem to manipulated extreme drought (45 days), N addition and their interaction, we measured leaf photosynthetic characteristics, aboveground phytomass on the community level and ecosystem C exchange in different treatments at the middle and the end of the drought period. The extreme drought treatment decreased the leaf net CO2 assimilation rate and ecosystem C exchange [gross ecosystem productivity (GEP), ecosystem respiration and net ecosystem CO2 exchange]. In contrast, the N addition treatment increased aboveground phytomass, GEP and net ecosystem CO2 exchange. The effects of N addition on the drought susceptibility of the L. chinensis meadow ecosystem varied with drought severity. The N addition treatment alleviated drought-induced suppression of CO2 exchange at the leaf and ecosystem levels in the middle of the drought period, whereas it exacerbated drought-induced suppression of the CO2 exchange and aboveground phytomass on the community level at the end of the drought period. Given that dominance by L. chinensis is a characteristic of the studied ecosystem, knowledge of the traits of L. chinensis and its response to multiple global change drivers will be crucial for predicting future ecosystem functions. Furthermore, increasing N deposition may affect the response of the L. chinensis meadow ecosystem to further droughts by increasing carbon allocation to roots and therefore root-shoot ratios.

Drought Sensitivity of the Carbon Isotope Composition of Leaf Dark-Respired CO2 in C3 (Leymus chinensis) and C4 (Chloris virgata and Hemarthria altissima) Grasses in Northeast China.

Whether photosynthetic pathway differences exist in the amplitude of nighttime variations in the carbon isotope composition of leaf dark-respired CO2 (δ13Cl) and respiratory apparent isotope fractionation relative to biomass (ΔR,biomass) in response to drought stress is unclear. These differences, if present, would be important for the partitioning of C3-C4 mixed ecosystem C fluxes. We measured δ13Cl, the δ13C of biomass and of potential respiratory substrates and leaf gas exchange in one C3 (Leymus chinensis) and two C4 (Chloris virgata and Hemarthria altissima) grasses during a manipulated drought period. For all studied grasses, δ13Cl decreased from 21:00 to 03:00 h. The magnitude of the nighttime shift in δ13Cl decreased with increasing drought stress. The δ13Cl values were correlated with the δ13C of respiratory substrates, whereas the magnitude of the nighttime shift in δ13Cl strongly depended on the daytime carbon assimilation rate and the range of nighttime variations in the respiratory substrate content. The ΔR,biomass in the C3 and C4 grasses varied in opposite directions with the intensification of the drought stress. The contribution of C4 plant-associated carbon flux is likely to be overestimated if carbon isotope signatures are used for the partitioning of ecosystem carbon exchange and the δ13C of biomass is used as a substitute for leaf dark-respired CO2. The detected drought sensitivities in δ13Cl and differences in respiratory apparent isotope fractionation between C3 and C4 grasses have marked implications for isotope partitioning studies at the ecosystem level.