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The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Some forages require significant amounts of water to grow, causing the dairy industry to be dependent on a limited resource. Feeding crop residues and feed coproducts in dairy rations may represent opportunities when alfalfa is not readily available, and may reduce the industry's use of water. A study using indirect calorimetry and 12 multiparous lactating Jersey cows (BW = 447.5 ± 43.7 kg; DIM = 71 ± 11 d, mean ± SD) was conducted to determine the effect of feeding dried distillers grains and solubles (DDGS) and straw in replacement of alfalfa hay on milk production and energy utilization. A triplicated 4 × 4 Latin square design was used to evaluate the replacement of alfalfa hay with a coproduct mixture (COP) of wheat straw and DDGS. Animals were blocked by milk yield and randomly assigned to 1 of 4 experimental treatments as follows (proportions on a DM basis): a control diet (CON) containing 18.2% of alfalfa hay, a low-coproduct diet (LCOP) that contained 8.1% COP, a medium-coproduct diet (MCOP) that contained 16.3% COP, and a high-coproduct diet (HCOP) that contained 24.3% COP. No differences were observed for daily DMI or milk yield (mean ± SEM), 19.5 kg ± 0.60 and 29.6 kg ± 0.91, respectively. A quadratic tendency was observed, where increasing inclusion of COP up to 16.3% maintained ECM and milk fat yield but decreased these when animals were fed 24.3% COP. Total methane production decreased linearly from 429.4 to 345.0 ± 22.8 L/d from CON to HCOP diets, respectively. The digestibility of CP increased linearly from 64.0% to 70.4% ± 0.95%, and N balance increased linearly from 43.3 to 90.7 ± 15.0 g/d in animals consuming CON to HCOP diets. Total time spent ruminating was lowest in animals consuming the HCOP diet. A linear increasing tendency in digestible and metabolizable energy of 2.92 to 3.02 ± 0.041 Mcal/kg and 2.58 to 2.70 ± 0.047 Mcal/kg was observed in animals consuming CON to HCOP. The proportion of metabolizable energy from digestible energy tended to linearly increase from 88.3 to 89.4 ± 0.454 when COP was added to the diet. Results of this study indicate that alfalfa hay with a mixture of straw and DDGS can maintain milk production and DMI, but the partial or full replacement of alfalfa with the COP mixture may result in differences in energy utilization, in part driven by effects on CH 4 reduction. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Variation in forage composition decreases the accuracy of diets delivered to dairy cows. However, variability of forages can be managed using a renewal reward model (RRM) and genetic algorithm (GA) to optimize sampling and monitoring practices for farm conditions. Specifically, use of quality control charts to monitor forage composition can identify changes in composition for which adjustment in the formulated diet will result in a better match of the nutrients delivered to cows. The objectives of this study were (1) to assess the use of a clustering algorithm to estimate the mean time (d) the process is stable or in control (T Stable) and the magnitude of the change in forage composition between stable periods (Δ Forage) for corn silage and alfalfa-grass silage, which are input parameters for the RRM; (2) to compare optimized farm-specific sampling practices (number of samples, sampling interval, and control limits [Δ Limit ]) using previously proposed defaults and our estimates for the T Stable and Δ Forage input parameters; and (3) to conduct a simulation study to compare the number of recommended diet changes under the proposed sampling and monitoring protocols. We estimated the T Stable and Δ Forage parameters for corn silage NDF and starch and alfalfa-grass silage NDF and CP using a k-means clustering approach applied to forage samples collected from 8 farms, 3×/wk during a 16-wk period. We compared 4 sampling and monitoring protocols that resulted from the 2 methods for estimating T Stable and Δ Forage (default values and our proposed method) and either optimizing only the control limit or optimizing the control limits, the number of samples, and the number of days between sampling. We simulated the outcomes of implementing the optimized monitoring protocols using a quality control chart for corn silage and alfalfa-grass silage of each farm. Estimates of T ^ S t a b l e and Δ ^ F o r a g e from the k-means clustering analysis were, respectively, shorter and larger than previously proposed default values. In the simulated quality control monitoring, larger Δ ^ F o r a g e estimates increased the optimized Δ Limit , resulting in fewer detected shifts in composition of forages, a lower frequency of false alarms, and a lower quality control cost ($/d). Recommended diet reformulation intervals from the simulated quality control analysis were specific for the type of forage and farm management practices. The median of the diet reformulation intervals for all farms using our optimal protocols was 14 d (quartile [Q] 1 = 8, Q 3 = 26) for corn silage and 16 d (Q 1 = 8, Q 3 = 26) for alfalfa-grass silage. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Variation in feed components contributes to variation and uncertainty of diets delivered to dairy cows. Forages often have a high inclusion rate (50% to 70% of DM fed) and variable composition, and thus are an important contributor to nutrient variability of delivered diets. Our objective was to quantify the variation and identify the main sources of variability in corn silage and alfalfa-grass haylage composition at harvest (fresh forage) and feed-out (fermented forage) on New York dairy farms. Corn silage and alfalfa-grass haylage were sampled on 8 New York commercial dairy farms during harvest in the summer and fall of 2020 and during their subsequent feed-out in the winter and spring of 2021. At harvest, a composite sample of fresh chopped forage of every 8-ha section of individual fields was collected from piles delivered for silo filling. During a 16-wk feed-out period, 2 independent samples of each forage were collected 3 times per week. The fields of origin of each forage sample during feed-out were identified and recorded using silo maps created at filling. A mixed-model analysis quantified the variance of corn silage DM, NDF, and starch and haylage DM, NDF, and CP content. Fixed effects included soil type, weather conditions, and management practices during harvest and feed-out, and random effects were farm, silo unit, field, and day. At harvest, between-farm variability was the largest source of variation for both corn silage and haylage, but within-farm sources of variation exceeded farm-to-farm variation for haylage at feed-out. At feed-out, haylage DM and NDF content had higher within-farm variability than corn silage. In contrast, corn silage starch showed higher within-farm variation at feed-out than haylage CP content. For DM content at feed-out, day-to-day variation was the most relevant source of within-farm variation for both forages. However, for the nutrient components at feed-out (NDF and CP for haylage; NDF and starch for corn silage) silo-to-silo variation was the largest source of variability. Weather conditions systematically explained a proportion of the farm-to-farm variability for both forages at harvest and feed-out. We concluded that because of the high farm-to-farm variation, corn silage and haylage must be sampled on individual farms. We also concluded that due to the high silo-to-silo variability, and the still significant day-to-day and field-to-field variability within-farm, corn silage and haylage should be sampled within individual silos to better capture changes in forage components at feed-out. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the potential consequences on calf intake, performance, behavior, ruminal microbiome, and ruminal epithelium development of combining the inclusion of chopped barley straw and alfalfa hay during the pre- and postweaning periods keeping concentrate to forage ratio constant among dietary treatments. Forty-five Holstein calves (44 ± 5.7 kg of body weight [BW] and 3 ± 1.5 d of age) individually penned were blocked by BW and randomly assigned to a common pellet concentrate fed ad libitum along with one of following forage feeding strategies: barley straw before and after weaning (S-S), barley straw before and alfalfa hay after weaning (S-A), or alfalfa hay before and after weaning (A-A). All calves received the same milk replacer regimen. Forage was supplied in a separated bucket at the rate of 7.5% (preweaning) and 15% (postweaning) of total solid feed intake of the previous day. Feed intake and BW were recorded daily and weekly, respectively. Rumen samples were obtained via a stomach tube at 53, 66, and 87 d and were composite in 3 samples of 5 animals each for subsequent rumen microbiome analysis. A rumen epithelium sample was taken by endoscopy at 90 d to assess gene expression of OCLN , CLDN4 , SLC9A1 , SLC9A3 , SLC16A1 , SLC16A4 , IL6 , and TGFB1. Data were analyzed with a mixed-effects model accounting for the fixed effects of block, forage, week of study, and their interaction, and calf as a random effect. The type of forage fed did not affect concentrate feed, forage, or total DM intake before weaning. However, S-A and A-A calves consumed less concentrate feed and S-A calves grew at a lower rate after weaning than S-S calves. Expression of the gene coding for SLC16A1 in the rumen epithelium was greatest in S-S among treatments. Rumen microbiome did not differ among treatments, while the relative abundance of Acidaminococcus and Selenomas genera increased, while Alloprevotella , Bifidobaterium , Olsenella , and Succiclasticum genera decreased with age. In conclusion, feeding barley straw before and after weaning was more effective than feeding alfalfa hay in promoting concentrate feed intake after weaning and fostering an increase in the expression of SLC16A1 in the rumen epithelium. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the inclusion of alfalfa grazing during 8 h continuous or partitioned in 2 separated sessions of 4 h after each milking, on nutrient intake, digestibility, ruminal fermentation, feeding behavior, milk production, milk composition, and milk fatty acid profile, in late-lactation cows fed a partial mixed ration (PMR). Twelve dairy cows (193 ± 83 d in milk, 584 ± 71 kg of body weight) were housed in individual outdoor pens and assigned to treatments according to a 3 × 3 Latin square design replicated 4 times. The treatments were as follows: (1) control (T0), cows were fed a total mixed ration (TMR) provided ad libitum 20.0% crude protein (CP), 32.2% neutral detergent fiber (NDF); (2) fed a diet combining a PMR which had the same ingredient composition as the TMR (60% of ad libitum intake) + 1 session of 8 h of pasture (T8), continuous grazing alfalfa (Medicago sativa ; 20.6% CP, 35.8% NDF) after the p.m. milking; and (3) PMR (60% of ad libitum intake) + 2 daily sessions of 4 h of access to pasture after each milking (T4+4). The experiment lasted 57 d and was divided into 3 periods of 19 d each. The first 12 d of each period was used for diet adaptation, and the last 7 d was used for data collection. No differences among treatments were observed for any of the productive variables, feeding efficiency, or purine derivatives excretion. Cows in T0 had greater intake and apparent digestibility of dry matter, organic matter, and nonfibrous carbohydrates compared with T4+4 and T8. Compared with T0, alfalfa grazing increased the concentration of C18:1 trans -11 and decreased those of C16:0 and C17:0 in milk fat. Cows in T4+4 consumed 1.1 more kg DM/d of alfalfa and N provided by alfalfa in the diet was 3 percentage points higher compared with T8 cows (266 vs. 229 g/d, respectively). In addition, T4+4 cows had a greater daily range of ruminal pH than T8 (0.73 vs. 0.93), and the highest concentrations of NH 3 -N were recorded during the a.m. grazing session while in T8 cows it occurred during the night. In conclusion, including 8 h of alfalfa grazing in T8 and T4+4 treatments allowed the substitution between 35.8 and 38.7% of the total dry matter intake (DMI) of a PMR (with a similar CP concentration to alfalfa) for pasture, maintaining milk solids production and increasing the C18:1 trans -11 of milk fat compared with a TMR in mid late–lactation cows. In an herbage plus PMR diet, splitting the 1 continuous grazing session of 8 h into 2 sessions of 4 h increased the proportion of energy and N provided by alfalfa pasture and reduced PMR intake, without modifying the total nutrient intake or productive performance of cows. [ABSTRACT FROM AUTHOR]

Polysaccharides have multiple pharmacological activities without toxicity or side effects. In the present study, the structure, antioxidant and lipid-lowering activities of alfalfa (Medicago sativa L.) polysaccharides (APS) were reported. Three alfalfa root polysaccharides (RAPS), RAPS-1, RAPS-2, and RAPS-3 were isolated, purified, and characterized. The structural analyses demonstrated that the homogeneous RAPS had molecular weights of 22.60, 26.95, and 38.25 kDa, respectively. Furthermore, RAPS-1 was composed of rhamnose, arabinose, xylose, glucuronic acid, glucose, and galactose with molar ratios of 10.25%, 12.53%, 6.58%, 4.56%, 15.74%, and 36.30%, RAPS-2 comprised rhamnose, arabinose, xylose, glucose, and galactose in the molar ratios of 24.07%, 14.48%, 1.24%, 15.05%, and 39.37%, while RAPS-3 was composed of rhamnose, arabinose, xylose, glucuronic acid, mannose, glucose, and galactose in the molar ratios of 31.61%, 11.78%, 1.93%, 5.49%, 3.18%, 14.56%, and 24.63%. The SEM analysis suggested that with the increase of the molecular weight of polysaccharides, its structure is more compact and the stability is stronger. In vitro , RAPS showed strong scavenging ability against DPPH and ABTS free radicals. In addition, RAPS had potent antioxidant activity and reduced the accumulation of lipids in oleic acid-induced NAFLD in HepG2 cells, with RAPS-1 being the most effective. Furthermore, RAPS-1 decreased MDA production, improved the activities of SOD and the total antioxidant capacity of cells, demonstrating that RAPS could effectively restrict lipid accumulation in HepG2 cells induced by oleic acid by restoring the antioxidant system. Western blot analysis indicated that the RAPS-1 can activate the Nrf2/HO-1 signaling pathway to relieve oxidative stress induced by oleic acid, thus playing a role in the treatment of NAFLD. [Display omitted] • Three new acidic polysaccharides were isolated from the roots of Medicago sativa L. • The polysaccharides exhibited considerable antioxidant and lipid-lowering activities. • The polysaccharides with antioxidant effect by activating Nrf2/HO-1 signaling pathway. [ABSTRACT FROM AUTHOR]

The objectives of this study were to determine the dry matter intake (DMI), urine pH, Ca concentration in blood, Ca output in urine, and frequency of hypocalcemia in pregnant and nonlactating dairy cows consuming diets containing different hays and acidogenic products during the prepartum period. Eighty pregnant and nonlactating Holstein cows approaching their second or greater calving were fed 1 of 4 experimental diets according to a 2 × 2 factorial arrangement of treatments during the prepartum period (21 d before calving). Diets included either grass hay (GH) or alfalfa hay (AH) and calcium chloride (CL) or polyhalite (PO) as the acidogenic products. All diets had a dietary cation-anion difference (DCAD) below −190 mEq/kg of dry matter (DM). Grass hay contained 75 g/kg crude protein, 749 g/kg neutral detergent fiber, 3.6 g/kg Ca, 0.9 g/kg Na, 18.8 g/kg K, 3.8 g/kg Cl, 1.5 g/kg S, and a cation-anion difference equal to 290 mEq/kg of DM. Alfalfa hay contained 196 g/kg crude protein, 456 g/kg neutral detergent fiber, 15.2 g/kg Ca, 1.6 g/kg Na, 25 g/kg K, 7.7 g/kg Cl, 3.2 g/kg S, and a cation-anion difference equal to 292 mEq/kg of DM. Cows consuming GH tended to consume more DM than cows consuming AH (11.6 vs. 10.8 kg/d) but DMI did not differ between acidogenic products. Urine pH decreased below 6.5 for all diets, although cows consuming the GHPO diet had the highest urine pH. The concentration of Ca in plasma decreased substantially around calving but neither hay type nor acidogenic product affected it. Urinary Ca output was lowest for cows consuming the GHPO diet. No associations existed between dietary treatments and the frequencies of normocalcemia and hypocalcemia. Under the conditions of this study, in which alfalfa and grass hays had similar cation-anion differences, we concluded that the inclusion of alfalfa hay in prepartum diets does not necessarily increase the frequency of hypocalcemia. The cation-anion difference of the alfalfa hay, more than the concentration of potassium alone, may be a key determinant of whether alfalfa hay fits in a prepartum feeding program for prepartum dairy cows. Further research should explore this relationship. [ABSTRACT FROM AUTHOR]

• Heavy metal pollution risk of desulfurized steel slag (DS) as soil amendment was evaluated both in field and pot experiments. • The addition of DS could increase the content of Cr but do not increase the contents of Cu, Zn, Cd, and Pb in soil. • DS application could alter bioavailable and chemical forms of heavy metals in soil and fulvic acid could further reduce the heavy metal pollution risk of DS in agricultural use. • The use of DS in agriculture is safe and feasible technology if it was applied properly. The by-product of wet flue gas desulfurization, desulfurized steel slag (DS), had chemical characteristics like natural gypsum that can be used to improve saline-sodic soil. However, contamination risk of heavy metals for cycling utilization of DS in agriculture was concerned mostly. Both pot and field experiments were conducted for evaluating the potential pollution risk of DS as the amendment of saline-sodic soil. Results showed that application of DS decreased the contents of Cd, Cu, Zn, and Pb, while significantly increasing chromium (Cr) content in DS-amended soils. The field experiment demonstrated that the migration of heavy metals (Cd, Zn, Cu, and Pb) in the soil profile was negligible. The application of DS at the dosage of 22.5–225 tons/ha significantly increased the Cr content in alfalfa (Medicago sativa L.) but lower than the national standard for feed in China (GB 13078-2017). DS altered the chemical fraction of heavy metals (Zn, Cu, and Pb), transferred exchangeable, reducible into oxidizable and residual forms in DS-amended soil. Application of DS combined with fulvic acid (FA) could effectively reduce the movement of heavy metals in soil and the accumulation of Cr in alfalfa. Based on our results, DS was a safe and feasible material for agricultural use and presented relatively little pollution risk of heavy metals. However, the results also showed that DS to a certain extent had a potential environmental risk of Cr if larger dosages of DS were used. [Display omitted] [ABSTRACT FROM AUTHOR]

The global population is forecast to reach around 9.7 billion by 2050, which means there will be an increased requirement for proteins in the human diet. Alfalfa leaf is a promising source of inexpensive protein for human consumption and food applications that is presently underutilized. Currently, the application of alfalfa leaf proteins (ALPs) in food formulations is limited due to their poor nutritional quality (anti-nutritional factors) and sensory attributes (bitterness and darkness). This review critically evaluates the application of alfalfa leaf as a potential protein source to meet global nutritional needs. Initially, the molecular composition and nutritional value of ALPs are discussed, and recent advances in their extraction and purification are highlighted. The techno-functional properties of ALPs are then covered, including their solubility, fluid binding (water and oil), emulsifying, foaming, and gelling properties. Finally, current challenges to the application of ALPs as a source of protein in the human diet are highlighted, as well as potential strategies to overcome these challenges. Alfalfa is commonly used as livestock feed owing to its high protein content. ALPs may also be used in food formulations, since it has a balanced amino acid composition and exhibits a range of functional attributes. However, post-harvest processing methods, such as blanching, isolation, and purification, must be optimized to create high-quality protein functional ingredients that are safe and nutritious. [Display omitted] • Alfalfa is a promising and sustainable source of plant-based protein. • Alfalfa protein may be used in food applications and human consumption. • The application of alfalfa leaf proteins in food is limited. • Post-harvest processing can improve the protein functionality and quality. [ABSTRACT FROM AUTHOR]

Reducing the dietary crude protein (CP) concentration can decrease the financial cost and lower the environmental impact of milk production. Two studies were conducted to examine the effects of reducing the dietary CP concentration on animal performance, nutrient digestibility, milk fatty acid (FA) profile, and nitrogen use efficiency (NUE; milk N/N intake) in dairy cows fed legume silage-based diets. Thirty-six multiparous Holstein-Friesian dairy cows that were 76 ± 14 (mean ± SD) days in milk and 698 ± 54 kg body weight were used in a 3 × 3 Latin square design in each of 2 studies, with 3 periods of 28 d. In study 1, cows were fed diets based on a 50:50 ratio of red clover to grass silage [dry matter (DM) basis] containing 1 of 3 dietary CP concentrations: high (H) = 175 g of CP/kg of DM; medium (M) = 165 g of CP/kg of DM; or low (L) = 150 g of CP/kg of DM. In study 2, cows were fed 175 g of CP/kg of DM with a 50:50 ratio of alfalfa to corn silage (H50) or 1 of 2 diets containing 150 g of CP/kg of DM with either a 50:50 (L50) or a 60:40 (L60) ratio of alfalfa to corn silage. Cows in both studies were fed a total mixed ration with a forage-to-concentrate ratio of 52:48 (DM basis). All diets were formulated to meet the MP requirements, except L (95% of MP requirements). In study 1, cows fed L ate 1.6 kg of DM/d less than those fed H or M, but milk yield was similar across treatments. Mean milk protein, fat, and lactose concentrations were not affected by diet. However, the apparent total-tract nutrient digestibility was decreased in cows fed L. The NUE was 5.7 percentage units higher in cows fed L than H. Feeding L also decreased milk and plasma urea concentrations by 4.4 mg/dL and 0.78 mmol/L, respectively. We found no effect of dietary treatment on the milk saturated or monounsaturated FA proportion, but the proportion of polyunsaturated FA was increased, and milk odd- and branched-chain FA decreased in cows fed L compared with H. In study 2, DM intake was 2 kg/d lower in cows receiving L50 than H50. Increasing the alfalfa content and feeding a low-CP diet (L60) did not alter DMI but decreased milk yield and milk protein concentration by 2 kg/d and 0.6 g/kg, respectively, compared with H50. Likewise, milk protein and lactose yield were decreased by 0.08 kg/d in cows receiving L60 versus H50. Diet had no effect on apparent nutrient digestibility. Feeding the low-CP diets compared with H50 increased the apparent NUE by approximately 5 percentage units and decreased milk and plasma urea concentrations by 7.2 mg/dL and 1.43 mmol/L, respectively. Dietary treatment did not alter milk FA profile except cis- 9, trans- 11 conjugated linoleic acid, which was higher in milk from cows receiving L60 compared with H50. We concluded that reducing CP concentration to around 150 g/kg of DM in red clover and grass or alfalfa and corn silage-based diets increases the apparent NUE and has little effect on nutrient digestibility or milk performance in dairy cows. [Display omitted] [ABSTRACT FROM AUTHOR]

As an antioxidant, SO 2 plays a critical role in maintaining the plant's redox balance. At the same time, Hg2+ as heavy metals, will accumulate in plants through soil, often disrupting physiological processes in plants. However, few studies have been reported to examine changes in Hg2+ and SO 2 in plants. Here, we developed a dual-channel fluorescent probe NYD, which can differentiate between the detection of Hg2+ and SO 2. The probe exhibits high sensitivity (LOD, 44 nM for Hg2+ and 26 nM for SO 2). When Hg2+ and HSO 3 − deprotect and attack the double bond by nucleophilic addition, significant emission of fluorescence at 475 nm and 574 nm with blue and red fluorescence respectively. In addition, NYD has the capability to detect the levels of Hg2+ and SO 2 in cells and alfalfa seedlings. Meanwhile, the findings indicated that alfalfa seedlings were exposed to Cd2+, which led to redox imbalance and increased endogenous SO 2 production. We anticipate that this research will contribute to a better understanding of the functions of SO 2 under oxidative stress induced by Cd2+. We developed a dual-channel fluorescent probe NYD, which detects Hg2+ and SO 2 by turning on the ICT effect and Michael addition reaction, respectively. The probe has a significant fluorescence enhancement at the emission wavelength of 574 nm and 475 nm, respectively, with a detection limit of 44 nM and 26 nM, and has good selectivity for Hg2+ and SO 2. We successfully applied it to the detection of Hg2+ and SO 2 levels in cells. Furthermore, Hg2+ and SO 2 in alfalfa seedlings were detected by imaging technique, and the up-regulation of SO 2 under Cd2+ stress was verified. [Display omitted] • NYD shown dual-emission wavelength for monitoring Hg2+ and SO 2 , respectively. • Hg2+ and SO 2 levels in HepG2 cell and plant were tracked using NYD. • NYD detected the up-regulation of SO 2 in alfalfa seedlings under Cd2+ stress. [ABSTRACT FROM AUTHOR]

It is well-established that maize (Zea mays L.) grown in extended rotations produces higher grain yields compared to maize grown in one- or two-phase rotations, even when nitrogen (N) is not limiting. Understanding the mechanisms driving this phenomenon, commonly referred to as 'the rotation effect', is important for designing cropping systems that use land and other resources efficiently. Differences in root systems can influence crop resource acquisition and therefore yield, but it is unknown if such differences play a role in the rotation effect. We hypothesized that maize grown in an extended rotation system exhibits a deeper root structure with less root mass compared to maize grown in a short rotation, and that these characteristics are correlated with differences in grain production. Using a long-term experiment established in 2001, we measured maize rooting depth across the growing season, root mass in 15 cm increments from 0 to 60 cm, and grain yields in the maize phase of two contrasting rotations: a 2-year rotation of maize/soybean (Glycine max [L.] Merr) using inorganic sources of nitrogen (N) and maximum tillage depths of 15 cm (hereafter the 'short rotation'), and a 4-year rotation of maize/soybean/oat (Avena sativa L.)-alfalfa (Medicago sativa L.)/alfalfa using a mix of organic and inorganic N sources and periodic inversion tillage to 25 cm (hereafter the 'extended rotation'). Additionally, we measured soil penetration resistance and soil moisture, and performed a growth analysis on aboveground maize biomass. From 2013 to 2020, maize grain yields in the extended rotation were equal to or significantly higher than in the short rotation, averaging 8 % greater across eight years (11.0 and 10.2 dry Mg ha−1, respectively). The timing (e.g., early season, late season) of the extended rotation's maize growth advantage was not consistent across years, but in all three seasons of root measurements (2019–2021) the maximum rooting depth of maize in the extended rotation was significantly deeper than in the short rotation by an average of 11 % (82 versus 76 cm, respectively). At physiological maturity, the two systems had similar amounts of root mass from 0 to 60 cm soil depth, but maize grown in the extended rotation invested significantly less of that mass (30 % compared to 47 %) into the soil surface layer (0 to 15 cm). The soil penetration resistances of the two systems differed in a manner consistent with the differing tillage regimes of the two rotations, however the patterns did not align with root differences. We posit that the extended rotation's 'deeper and steeper' maize root patterns did not guarantee higher maize yields, but rather bestowed the plant with more flexibility in resource acquisition which, in certain conditions, resulted in higher grain yields compared to maize grown in the short rotation. To our knowledge, this is the first report attempting to mechanistically link rooting patterns with plant growth in the context of the 'rotation effect.' This study enhances our understanding of how cropping system histories impact yields, and provides new data on yields and roots, both of which are highly relevant for sustainable intensification. While the present study focused on physical measurements, it suggests that more detailed exploration of how biological drivers impact root architecture is needed to gain a mechanistic understanding of the 'rotation effect.' • Maize grain yields in the four-year rotation were 8 % higher than in the two-year. • Maize roots extended 11 % deeper in the four-year compared with the two-year. • Maize had less shallow (0–15 cm) root biomass in the four-year compared with the two-year. • Four-year rotation maize roots were 'deeper and steeper' compared with the two-year. • Differences were not explained by physical soil measurements. [ABSTRACT FROM AUTHOR]

Seed aging poses a significant challenge to agronomic production and germplasm conservation. Reactive oxygen species (ROS) are highly involved in the aging process. However, dynamic response of germination characteristics and antioxidant system to seed aging are not yet very clear. This study explored the potential physiological mechanisms responsible for the reduced and rapid loss of seed vigor in alfalfa, and identified key genes regulating seed vigor. The germination percentage exhibited a decreased trend with the prolongation of aging duration. From 16 to 32 days of aging, the antioxidant enzyme activities of SOD, POD, CAT, DHAR and MDHAR declined significantly, which lead to the disruption of ROS balance and a significant increase in ROS levels, exacerbating seed aging. Based on transcriptome, 29 differentially expressed genes (DEGs) including SOD1 , APX-2 and GST-7 within the ROS scavenging system showed a significantly down-regulated expression trend at aging of 16 and 24 days, indicating the abnormal function of antioxidant metabolism. Furthermore, some related genes including ATPF1B , ATPeF0C-3 , NDUFS1 , NDUFS3 and ND2 in the mitochondrial ETC exhibited a downturn following seed aging, which would result in the losing of seed vigor. This study has uncovered a significant array of potential target genes within the seed antioxidant system and mitochondrial ETC. These discoveries offer a wider lens for delving into the molecular regulatory mechanisms of seed aging. Further research is crucial to comprehensively elucidate the precise pathways through which these pivotal genes regulate seed vigor. • Response patterns of antioxidant systems were investigated in aging alfalfa seed. • The germination percentage exhibited a decrease with the prolongation of aging duration. • ROS suppressed the germination of aging alfalfa seeds. • 29 differentially expressed genes within the ROS scavenging system were identified. [ABSTRACT FROM AUTHOR]

Soil salinization negatively affects plant growth and threatens food security. Halotolerant plant growth-promoting bacteria (PGPB) can alleviate salt stress in plants via diverse mechanisms. In the present study, we isolated salt-tolerant bacteria with phosphate-solubilizing abilities from the rhizosphere of Salix linearistipularis , a halophyte distributed in saline-alkali soils. Strain A103 showed high phosphate solubilization activity and was identified as Enterobacter asburiae based on genome analysis. In addition, it can produce indole-3-acetic acid (IAA), siderophores, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Genome mining has also revealed the presence of several functional genes involved in the promotion of plant growth. Inoculation with A103 markedly improved alfalfa growth in the presence of 100 mM NaHCO 3. Under alkali stress, the shoot and root dry weights after bacterial inoculation improved by 42.9 % and 21.9 %, respectively. Meanwhile, there was a 35.9–37.1 % increase in the shoot and root lengths after treatment with A103 compared to the NaHCO 3 -treated group. Soluble sugar content, peroxidase and catalase activities increased in A103-inoculated alfalfa under alkaline stress. A significant decrease in the malondialdehyde content was observed after treatment with strain A103. Metabolomic analysis indicated that strain A103 positively regulated alkali tolerance in alfalfa through the accumulation of metabolites, such as homocarnosine, panthenol, and sorbitol, which could reduce oxidative damage and act as osmolytes. These results suggest that halophytes are valuable resources for bioprospecting halotolerant beneficial bacteria and that the application of halotolerant growth-promoting bacteria is a natural and efficient strategy for developing sustainable agriculture. [Display omitted] • Enterobacter asburiae A103 was isolated from the halophyte Salix linearistipularis. • Strain A103 possesses multiple plant growth-promoting traits. • Genes related to plant growth promotion were identified in strain A103 genome. • Strain A103 could efficiently stimulate alfalfa growth under alkaline stress. • Strain A103 inoculation led to high accumulation of homocarnosine and panthenol. [ABSTRACT FROM AUTHOR]

• Bis-Triazole amended S-heterocyclic based Schiff base was synthesized via click chemistry and simple condensation reaction. • Molecular docking was performed on synthesized compounds with cytochrome p-450 oxidase. • The efficacy of compound-4 was examined on the seedling growth of Zea mays and Medicago sativa. • UV–Vis. Absorbance spectra of compound-4 suggest that it selectively detect Co(II) metal ion. The present investigation supports the synthesis of Bis-Triazole allied S-heterocyclic based Schiff base via click chemistry and simple condensation approach. The molecular docking was performed on both the compounds which interprets that after the addition of Schiff base to Bis-Triazole moiety it showed better binding affinity for cytochrome p450 oxidase. Further the effect of compound-4 was seen on the seedling growth of Zea mays and Medicago sativa. When applied to Zea mays, compound-4 demonstrated its best benefits at a concentration of 0.5 mg L −1, whereas 10 mg L −1 of Medicago sativa displayed promising results. In addition, UV–Vis. Absorbance studies revealed that compound-4 selectively detects Co(II) in presence of several other metal ions, with good value of limit of detection and association constant. [ABSTRACT FROM AUTHOR]

[Display omitted] • Red-edge and SWIR regions are crucial for accurately predicting alfalfa NDF and ADF. • Vegetation indices based on three bands can improve the accuracy of NDF and ADF. • Sentinel-2 multispectral data has great potential for estimating NDF and ADF. Alfalfa, a high-quality forage, has good palatability and nutritional value. Neutral detergent fiber (NDF) and acid detergent fiber (ADF) are both key indicators of alfalfa quality. However, the uncertainties in existing studies regarding the sensitive bands and inversion mechanism for NDF and ADF contents estimations have limited the application of high-precision remote sensing-based inversion. In this study, using hyperspectral and Sentinel-2 (S2) multispectral data of cultivated alfalfa in the Hexi Corridor region from 2020 to 2022, we analyze the characteristic spectral band and vegetation indices (VIs) required to estimate the NDF and ADF contents of alfalfa. The key conclusions are as follows. (1) The sensitive bands selected using ASD hyperspectral data are mainly in the blue, green, red-edge, and short-wave infrared (SWIR) regions, while the sensitive bands based on S2 data cover a broader range between the blue and SWIR regions. (2) Among the 21 NDF and 21 ADF models based on ASD data in this study, the optimal models are both artificial neural network (ANN) models constructed by VIs (R2 of 0.80 for both, RMSEs of 2.27% and 1.75% and mean absolute errors (MAEs) of 1.77% and 1.38% for NDF and ADF, respectively). For the S2 data, the optimal models are also ANN-based and constructed using VIs (with R2 values of 0.66 and 0.72, RMSEs of 3.06% and 2.24%, and MAEs of 2.50% and 1.79% for NDF and ADF, respectively. (3) The inversion results using the optimal model indicate that the proportion of alfalfa area in the typical study area with NDF and ADF contents characterized by a supreme grade is greater than 60%. Overall, both the ASD hyperspectral and S2 multispectral data can accurately predict alfalfa NDF and ADF contents. This approach provides an effective technical means by which the management of local alfalfa production may be guided. [ABSTRACT FROM AUTHOR]

Shade is a stressful factor for most plants, leading to both morphological and physiological changes, and often resulting in increased susceptibility to diseases and pathogen attacks. Our study revealed that the isoflavonoid synthesis pathway was inhibited in alfalfa under shade, resulting in a significant reduction in disease resistance. Overexpression of MsIFS1 , a switch regulator in isoflavonoid synthesis, led to a notable increase in endogenous isoflavonoids and enhanced resistance to Pseudomonas syringae pv. tomato DC3000 (Pst. DC3000). Conversely, MsIFS1 -RNAi had the opposite effect. Yeast one-hybrid (Y1H) assays revealed that the shade-responsive transcription factor MsWRKY41 could directly bind to the MsIFS1 promoter. This interaction was confirmed through Dual-Luciferase Reporter (Dual-LUC) and Chromatin Immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) assays, both in vitro and in vivo. Overexpression of MsWRKY41 not only enhanced alfalfa's resistance to Pst. DC3000 but also promoted the accumulation of isoflavonoids. Additionally, yeast two-hybrid (Y2H) assays showed that neither MsWRKY41 nor MsIFS1 physically interacted with the Type III effector (T3SE) HopZ1 secreted by Pst. DC3000, suggesting that the MsWRKY41-MsIFS1 module is not a direct target of HopZ1. These findings provide valuable theoretical insights and genetic resources for the development of shade-tolerant alfalfa with enhanced disease resistance. • Shade leads to reduced disease resistance in alfalfa. • Isoflavonoid synthesis is regulated by light. • MsIFS1 affects disease resistance by influencing Isoflavonoid metabolism in alfalfa. [ABSTRACT FROM AUTHOR]

High pH saline-alkali stress, mainly NaHCO 3 , limited the development of animal husbandry in Songnen Plain. Ion imbalance and reactive oxygen species (ROS) metabolism disorder caused by saline-alkali stress inhibited plant growth. In this study, we compared the differences in ion absorption, transport and ROS metabolism between saline-tolerant alfalfa (ZD) and saline-sensitive alfalfa (ZM) under NaHCO 3 stress using physiology and transcripomics techniques. WGCNA analysis identified key genes associated with NaHCO 3 stress-induced changes. NaHCO 3 stress inhibited the absorption of K+ and Mg2+, but activated Ca2+ signal. Furthermore, ZD maintained higher K+, Mg2+ and Ca2+ contents and the K+/Na+ ratio than ZM, this is mainly related to the higher expression of proteins or channel-encoding genes involved in ion absorption and transport in ZD. Antioxidant enzyme systems can be activated in response to NaHCO 3 stress. Peroxidase (EC 1.11.1.6), catalase (EC 1.11.1.7) and glutathione transferase (EC 2.5.1.18) activities were higher in ZD than ZM, and most genes encoding the relevant enzymes also demonstrated a stronger up-regulation trend in ZD. Although NaHCO 3 stress inhibited Trx-Prx pathway, ZD related enzymes and their genes were also inhibited less than ZM. WGCNA results identified many genes involved in ion absorption, transport and antioxidant systems that play an important role in NaHCO 3 stress adaptation. Collectively, ZD has the stronger ion homeostasis regulation and ROS scavenging ability, so it's more resistant to NaHCO 3. The results provide theoretical guidance for further understanding of the molecular mechanism of NaHCO 3 resistance and provide potential genes for research to improve saline-alkali tolerance in alfalfa. [Display omitted] • NaHCO 3 stress inhibited the absorption of K+ and Mg2+, resulting in a lower K+/Na+ ratio. • Salt-tolerant alfalfa had higher Mg2+, Ca2+ contents and the K+/Na+ ratio than saline-sensitive alfalfa under NaHCO 3 stress. • NaHCO 3 stress reduced ROS accumulation through AsA-GSH cycle but inhibited the Trx-Prx pathway in alfalfa. • Under NaHCO 3 stress, ZD maintained higher antioxidant enzyme activities and related coding genes expression levels in than ZM. [ABSTRACT FROM AUTHOR]

The use of alfalfa in rotation with intensive crops is common practice to mitigate the physical and chemical issues arising from intensive farming practices. However, there is a dearth of studies on this practice. Given the current concern regarding climate change and the significant impact agriculture has on greenhouse gas (GHG) emissions, understanding the emissions associated with this practice, as well as the most suitable soil and crop management techniques for their mitigation, is of paramount importance. The present study aimed to (i) quantify emissions of N 2 O, CO 2 and CH 4 in an alfalfa crop following a maize cropping scenario; (ii) to determine which tillage system generates the lowest GHG emissions, and; (iii) to determine how N fertilisation from a preceding intensive maize crop affects GHG emissions during alfalfa cropping period. A three-year field experiment (2019, 2020 and 2021) was conducted to assess the emissions of N 2 O, CO 2 and CH 4 from alfalfa cultivation following a three-year period of irrigated maize. Two soil management practices (no-tillage and conventional tillage) were implemented during both the maize cropping period and the alfalfa establishment. Additionally, the nitrogen (N) fertilisation rates applied to the preceding maize crop were included as a treatment (0, 200, and 400 kg N ha⁻¹, corresponding to zero, medium, and high fertilisation levels, respectively) in a randomized block design with two factors. Emissions of N 2 O in alfalfa ranged from 0.05 to 0.32 mg N 2 O-N m⁻² day⁻¹, being significantly higher only during first month of sampling in the treatments that had received fertilisation. CO 2 emissions ranged from 1158 to 4258 mg CO 2 -C m⁻² day⁻¹. Year-average CH 4 fluxes were −0.27 g C ha⁻¹ day⁻¹. The average total dry matter produced by alfalfa was 17700 kg ha⁻¹ year⁻¹, being higher for the no-tillage treatment, though significantly so only during first month of sampling. Under Mediterranean conditions, the tillage system and mineral N fertilizer rates have a relative effect on greenhouse gas emissions during the alfalfa cropping period. Plots without N fertilization initially produced lower N 2 O emissions and higher total dry matter, resulting in the lowest scaled emissions. For the tillage treatment, no significant differences were found in emission dynamics, which may be due to the fact that alfalfa does not involve soil disturbance, leading to a homogenization of the treatments. However, the NT treatment showed lower scaled emissions due to higher yields in the first year. Therefore, alfalfa cultivation is characterized by low GHG emissions, high yields, and a notable capacity to mitigate the negative effects of previous intensive crops. This study provides data on GHG emissions during alfalfa cropping in the typical Maize-alfalfa crop rotation under Mediterranean irrigated systems, which is useful for new agricultural policies aimed at reducing pollution in the agricultural sector. Additionally, it demonstrates the capability of this crop to mitigate adverse agronomic and environmental conditions caused by preceding intensive farming practices. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) consumes a large amount of soil inorganic nitrogen (N) but can supply ample rhizosphere deposited N to subsequent crops. Therefore, N fertilizer application levels should be optimized for corn under long-term alfalfa-corn (AC) rotation system to achieve high yield and N use efficiency. The present study assessed the yield and water and N use efficiency of corn under N fertilizer application in a long-term AC cropping system and optimized the corn N fertilizer application level using the Agricultural Production Systems sIMulator (APSIM). APSIM was calibrated and validated utilizing the experimental datasets of yield, aboveground biomass, plant N uptake, soil water storage, and inorganic N at 0−140 cm soil layer during corn growth with four N fertilizer treatments (0, 130, 195, and 260 kg N ha−1), which were collected from a six-year-old alfalfa field experiment carried out in Lishu County (Jilin Province, China) from 2020 to 2022; the field experiment was initiated in 2014. The validated APSIM was then utilized to simulate the long-term (1981−2020) characteristics of crop and soil under different corn N fertilizer application levels in a continuous corn (CC) cropping system and different alfalfa-corn rotation systems (one, two, three, four, and five years of alfalfa followed by two years of corn; 1A2C, 2A2C, 3A2C, 4A2C, 5A2C). The simulated N treatments included 0−300 kg N ha−1 range with an increment of 30 kg N ha−1. Model evaluation revealed that APSIM effectively captured the dynamics of the crop, soil water, and soil inorganic N during corn cultivation following alfalfa at four N fertilizer application levels. The normalized root-mean-square errors between the observed and simulated values under different treatments were less than 30 %. Alfalfa had legacy effects on the soil water and soil N mineralization (N min) of subsequent first-year corn, which ensured the corn yield following alfalfa. The first-year net N min in the soil with corn following alfalfa increased by 140 % (65 %−268 %) compared to the CC cropping system. Alfalfa planting also increased the 0−140 cm soil inorganic N before sowing (N sow) by 351 % (292 %−463 %) for the subsequent corn with no N fertilizer application and the 0−140 cm soil water storage before sowing by 22 % for the subsequent corn with relatively high N fertilizer application (300 kg N ha−1) compared to the CC cropping system. The highest yield and N use efficiency could be achieved by applying 90 kg N ha−1 N fertilizer for 1A2C/2A2C/3A2C rotation systems and 60 kg N ha−1 N fertilizer for 4A2C/5A2C rotation systems to the first-year corn following alfalfa. However, the N fertilizer requirement of the second-year corn following alfalfa under AC rotation systems was the same as that under the CC cropping system (150 kg N ha−1). The present study's findings indicate 60 kg N ha−1 N fertilizer application to the first-year corn following alfalfa under 1A2C/2A2C/3A2C rotation systems and 90 kg N ha−1 under 4A2C/5A2C rotation systems. The study provides a deeper understanding of alfalfa's effects on subsequent corn and a guidance for planning N fertilizer management in corn production following alfalfa. • 1st-yr corn following 1–3- and 4–5-year-old alfalfa required 60 and 90 kg ha−1 nitrogen (N) fertilizer, respectively. • Net N mineralization in the first-year corn following alfalfa improved by 140 % compared to continuous corn (CC). • Initial inorganic N (0−140 cm) in corn with no N fertilization following alfalfa was 351 % higher than that in CC. [ABSTRACT FROM AUTHOR]

Accurate estimation of soil moisture content (SMC) is essential for optimizing irrigation schedules and identifying drought-tolerant varieties. The integration of unmanned aerial vehicles (UAVs) with advanced sensors provides a novel method for monitoring SMC with high flexibility, resolution, and performance. This study utilized UAVs to capture RGB, multispectral, and thermal imagery of alfalfa (Medicago sativa L.) at the Linze Grassland Agricultural Experiment Station, Lanzhou University, and to evaluate the potential of fusing multimodal UAV data for SMC estimation in the root zone of densely and uniformly distributed leafy plants, using alfalfa as a case study, within a deep learning framework. Results showed that combining multimodal data—encompassing canopy spectral, structural, thermal, and textural information—significantly improved SMC estimation accuracy. Among the four regression models evaluated—partial least squares (PLSR), support vector machine (SVM), random forest (RF), and deep neural network (DNN)—the DNN model achieved the highest accuracy in overall multimodal data fusion, with a coefficient of determination (R 2) of 0.72 and a root mean square error (RMSE) of 4.98%. It demonstrated good predictive performance for both full and deficit irrigation scenarios, with R 2 values of 0.74 and 0.75, respectively. The DNN model also provided reliable SMC estimates across the three alfalfa canopy types, with R 2 values of 0.72, 0.74, and 0.58, respectively. Moreover, it exhibited superior accuracy under both irrigation regimes and demonstrated strong spatial adaptability, characterized by low spatial dependence and autocorrelation. In conclusion, the DNN model based on UAV-derived multimodal data fusion offers a reliable and robust approach for SMC estimation, providing valuable insights for irrigation management at farmland-scale. [Display omitted] ● Soil moisture content (SMC) in alfalfa root-zone was estimated using UAV-based multi-senor data fusion. ● Multimodal data fusion weakened spatial clustering of estimating SMC errors and enhanced the spatial adaptability. ● UAV-based multi-senor data fusion and deep learning improved accuracy of SMC estimation. [ABSTRACT FROM AUTHOR]

The FtsH (Filamentous temperature sensitive H) proteases, known for their crucial roles in protein quality control and maintaining the integrity of photosynthetic machinery, have emerged as key regulators of stress responses in plants. Our previous study revealed the overexpression of MsFtsH8 , an FtsH gene from alfalfa (Medicago sativa L.), confers salt stress tolerance to the plant. By comparing the proteomic profiles of MsFtsH8 -overexpressing alfalfa and wild type under salt stress conditions, we elucidate the molecular pathways underlying MsFtsH8-mediated salt stress resilience. We identified 730 differentially expressed proteins (DEPs) in MsFtsH8 -overexpressing alfalfa under salt stress, compared to 498 DEPs in wild type alfalfa under the same growth condition. Our results reveal significant alterations in the expression of proteins involved in the photosynthetic system, consistent with the chloroplast subcellular localization of MsFtsH8. Specifically, MsFtsH8 overexpression stabilizes key components of Photosystem II (PSII) and enhances electron transport processes, leading to increased photosynthetic efficiency and oxidative photodamage repair capacity under salt stress. Moreover, MsFtsH8 -overexpressing alfalfa exhibits elevated levels of antioxidative enzymes, further mitigating oxidative damage induced by high salinity. These findings deepen our understanding of the regulatory role of MsFtsH8 in salt stress response and highlight its potential for improving crop resilience under adverse environmental conditions. • Overexpression of MsFtsH8 stabilizes the photosystem II (PSII) system under salt stress. • Overexpression of MsFtsH8 enhances oxygen-evolving complex (OEC) functionality under salt stress. • Overexpression of MsFtsH8 mitigates oxidative damage with continuous electron flow under salt stress. [ABSTRACT FROM AUTHOR]

• Pellets and capsules of Medicago sativa can enhance growth and biotic stress resistance in okra and cowpea plants. • 3 and 5 % leaf pellets and capsules both increased inhibition of root rot colonization against applied pathogens. • Utilization of pellets and capsules could be an effective bio-stimulant for enhancing okra and cowpea disease resistance and physiological reactions. Medicago sativa L. is recognized for its economic value as a forage crop and can serve as an effective bio-stimulant to enhance soil fertility and plant productivity. In present study, leaf of M. sativa powdered were prepared in form of pellets and capsules and administered to the soil with different numbers (1, 3 and 5 respectively) to assess their impact on plant physiological performance and disease resistance against root rot fungi including: Fusarium oxysporum, Rhizoctonia solani, and Macrophomina phaseolina. Plants treated with three number of pellets and capsules showed notable improvements in growth parameters including shoot length, shoot weight, root length, root weight, and nodule count for both okra and cowpea. Furthermore, the application of M. sativa led to significant inhibition of root rot colonization by F. oxysporum, R. solani, and M. phaseolina in okra and cowpea plants. On the contrary, plants treated with five pellets and capsules exhibited enhancements in shoot length, shoot weight, root length, root weight, and notable inhibition of root rot colonization by F. oxysporum, R. solani, and M. phaseolina in both plants. Near-infrared spectroscopy analysis revealed that the increase in 5 pellets, as opposed to capsules, led to increased levels of protein, the chlorophyll content index, and moisture compared to the three pellets. Conversely, the application of three pellets resulted in the highest ash, fat, fiber, and starch content in okra plants compared to other treatments. In conclusion, the application of three pellets proves to be an effective bio-stimulant for enhancing plant biomass, disease resistance, and overall plant performance, thereby contributing to the maintenance of food security. [Display omitted] [ABSTRACT FROM AUTHOR]

Alfalfa often suffers from low temperature during spring rejuvenation, so it is important to improve the cold tolerance of alfalfa leaves for its smooth rejuvenation, and the alternative pathway (AP) could effectively improve the plant's tolerance. In this study, the contribution of AP on spring rejuvenation of alfalfa was investigated in Xinmu No.4 and Gannong No.5 with different fall dormancy levels. Though the protein and AP capacity were decreased during the rejuvenation, the ratio of AP/TP were increased in two alfalfa varieties, compared to those in alfalfa before overwintering. This indicated that AP had positive response to alfalfa rejuvenation. The limitation of AP significantly affected the leaf length, leaf width and growth rate of greening alfalfa, showing that AP played an important role in alfalfa rejuvenation. Inhibition of AP resulted in a significant decrease in Pn, Ci, Gs and stomatal structure deformity, suggestion that AP affected photosynthesis by influencing stomatal development during rejuvenation. AP reduces oxidative damage to PSII core protein repair in alfalfa leaves and optimizes photosynthesis by up-regulating NADP-MDH activity, decreasing the accumulation of excess reducing power in the chloroplasts, and by increasing SOD and POD activities and decreasing the accumulation of hydrogen peroxide. The higher proportion of AP keeps it more tolerant to low temperature for rejuvenation in Xinmu No.4 with a lower fall dormancy level. [ABSTRACT FROM AUTHOR]

There is an urge to optimise the sustainability and resilience of grasslands, especially in light of the extreme weather risks posed by climate change. Plant sown diversity could contribute to mitigating these risks by improving resource use efficiency as well as reducing reliance on fertilisation. We established a field experiment to determine forage swards' performance in terms of water use, nitrogen (N) use, and dry matter (DM) yield in a semi-arid Mediterranean area which faced a severe drought period. Most variables were tested over three growing seasons, whilst DM yield had an extra year of sampling in which two levels of N fertilisation were tested. Following the Generalised Diversity-Interactions modelling approach, we manipulated the composition and evenness of forage swards including a grass (Festuca arundinacea), a legume (Medicago sativa), and a non-legume forb (Cichorium intybus). Communities were grown as monocultures or mixtures of three species according to a simplex design, which allowed us to estimate separately the two components of the diversity effect: the individual species effects and that due to species interactions. Models were tested both with sown proportions (P i0) and realised proportions (P iR). The P i0 models indicated a major beneficial effect of mixtures in contrast to the average monoculture performance for all studied variables. In contrast, P iR models revealed that some of the effects in P i0 models are indeed explained by the growing proportion of M. sativa over time. However, beneficial interaction effects are still found in P iR models, especially concerning N 2 fixation (with δ15N as proxy), which was maintained regardless of the drought stress level experienced by the swards. All things considered, mixed swards merit attention as a valuable nature-based solution that can optimise N management (by boosting N 2 input through fixation) as well as water use. This would result in well-nurtured swards with a better ability to cope with drought stress and maintain productivity. [Display omitted] • Sown forage mixtures showed major benefits in terms of resource use and yield. • Some mixture benefits were mediated by Medicago sativa growing dominance over time. • Drought induced punctual negative species interactions that reduced yield. • δ 13C indicated greater water use in mixtures at a critical water-limited period. • Positive mixture effects on N 2 -fixation persisted regardless of water stress. [ABSTRACT FROM AUTHOR]

Downy mildew disease, which is caused by Peronospora aestivalis (Mastigomycotina: Peronosporaceae) and the spotted alfalfa aphid Therioaphis trifolii (Hemiptera: Aphididae), affects Medicago sativa (alfalfa) production. P. aestivalis and T. trifolii frequently occur together under natural conditions. In this study, alfalfa- P. aestivalis - T. trifolii were used as the research objects to explore the effects of different degrees of P. aestivalis infection on the growth and reproduction of T. trifolii , and the life table of T. trifolii population was constructed based on the fecundity parameters. The results revealed that alfalfa downy mildew infection had a significant effect on T. trifolii population. T. trifolii fed on mildly infected alfalfa plants exhibited the longest life span (18.9 d) and the highest number of nymphs (30.6) compared to those fed on alfalfa in other treatments. In terms of population life table parameters, the intrinsic rate of increase (0.626) and finite rate of increase (1.869) of T. trifolii fed on moderately infected alfalfa were the highest, and the generation time (8.310) and population doubling time (1.108) were the shortest, indicating that moderately infected alfalfa are conducive to T. trifolii population development. This study provides insights into the effects of alfalfa downy mildew on the main reproductive parameters and population of T. trifolii on M. sativa. • Therioaphis trifolii and Peronospora aestivalis often occur on alfalfa leaves at the same time. • The occurrence of Peronospora aestivalis can effectively affect the growth and development of Therioaphis trifolii. • There were differences in the growth and development of Therioaphis trifolii under different degrees of infection. • Feeding on moderately infected alfalfa is more beneficial to the reproduction of Therioaphis trifolii population. [ABSTRACT FROM AUTHOR]

Numerous studies have investigated seed aging, with a particular emphasis on the involvement of reactive oxygen species. Reactive oxygen species diffuse into the nucleus and damage telomeres, resulting in loss of genetic integrity. Telomerase reverse transcriptase (TERT) plays an essential role in maintaining plant genomic stability. Genome-wide analyses of TERT genes in alfalfa (Medicago sativa) have not yet been conducted, leaving a gap in our understanding of the mechanisms underlying seed aging associated with TERT genes. In this study, four MsTERT genes were identified in the alfalfa genome. The expression profiles of the four MsTERT genes during seed germination indicated that MS. gene79077 was significantly upregulated by seed aging. Transgenic seeds overexpressing MS. gene79077 in Arabidopsis exhibited enhanced tolerance to seed aging by reducing the levels of H 2 O 2 and increasing telomere length and telomerase activity. Furthermore, transcript profiling of aging-treated Arabidopsis wild-type and overexpressing seeds showed an aging response in genes related to glutathione-dependent detoxification and antioxidant defense pathways. These results revealed that MS. gene79077 conferred Arabidopsis seed-aging tolerance via modulation of antioxidant defense and telomere homeostasis. This study provides a new way to understand stress-responsive MsTERT genes for the potential genetic improvement of seed vigor. [ABSTRACT FROM AUTHOR]

Extensive bodies of research are dedicated to the study of seed aging with a particular focus on the roles of reactive oxygen species (ROS), and the ensuing oxidative damage during storage, as a primary cause of seed vigor decreasing. ROS diffuse to the nucleus and damage the telomeres, resulting in a loss of genetic integrity. Protection of telomeres 1 (POT1) is a telomeric protein that binds to the telomere region, and plays an essential role in maintaining genomic stability in plants. In this study, there were totally four MsPOT1 genes obtained from alfalfa genome. Expression analysis of four MsPOT1 genes in germinated seed presented the different expressions. Four MsPOT1 genes displayed high expression levels at the early stage of seed germination, Among the four POT1 genes, it was found that MS. gene040108 was significantly up-regulated in the early germination stage of CK seeds, but down-regulated in aged seeds. RT-qPCR assays and RNA-seq data revealed that MsPOT1-X gene was significantly induced by seed aging treatment. Transgenic seeds overexpressing MsPOT1-X gene in Arabidopsis thaliana and Medicago trunctula exhibited enhanced seed vigor, telomere length, telomerase activity associated with reduced H 2 O 2 content. These results would provide a new way to understand aging stress-responsive MsPOT1 genes for genetic improvement of seed vigor. Although a key gene regulating seed vigor was identified in this study, the specific mechanism of MsPOT1-X gene regulating seed vigor needs to be further explored. [ABSTRACT FROM AUTHOR]

Microbes play a key role in mediating soil nitrogen (N) cycling in agroecosystems. However, it remains unknown how N management practices affect the taxonomic and functional structure of soil N-cycling microbes, and their community assembly and co-occurrence networks in karst agroecosystems. Here, we conducted a field experiment to examine the effects of mineral N addition (+N) and legume (Medicago sativa) intercropping (+L) on soil N-cycling functional taxa and genes in a karst forage (Broussonetia papyrifera) agroecosystem. Results showed that compared to the control and +L treatment, mineral N addition significantly increased the functional gene diversity of nitrification-related microbes and the abundance of hao gene, but slightly reduced the abundances of nifD and nifH genes related to N fixation by 33.3−56.0 %. The abundance of nifK gene was 3.7-fold higher in the +L treatment than in the control. The assembly of microbial communities involved in ammonification, assimilatory nitrate reduction (ANR) and denitrification was controlled by a homogeneous selection process. Stochastic processes played a dominant role in shaping the communities related to nitrification, dissimilatory nitrate reduction (DNR), N fixation, and N assimilation. High soil pH and total N stimulated microbial N assimilation and the related gene abundance (e.g., GDH2), but suppressed the abundances of genes involved in N fixation. Both mineral N addition and legume intercropping significantly increased the niche breadth of the whole community and the functional groups related to denitrification, ANR, DNR and N assimilation. Actinobacteria related to N assimilation dominated the co-occurrence networks across the treatments. Compared to the control, the network robustness was significantly increased in the +N and +L treatments. Our findings indicate that there are distinct responses to the two N management practices among N-cycling functional groups and highlight the importance of N fertilization in increasing the niche breadth of N-cycling microbes and stabilizing their co-occurrence network in a karst agroecosystem. [Display omitted] • Mineral nitrogen (N) addition significantly increased the functional gene diversity of nitrification-related microbes. • High soil pH and total N promoted microbial N assimilation, but suppressed the N fixation potential. • The deterministic and stochastic processes co-drove the assembly of soil N-cycling microbial communities. • N fertilization increased the niche breadth of the whole N-cycling microbial community and stabilized the co-occurrence network. [ABSTRACT FROM AUTHOR]

Saline-alkali stress is one of the main abiotic stresses that limits plant growth. Salt stress has been widely studied, but alkaline salt degradation caused by NaHCO 3 has rarely been investigated. In the present study, the alfalfa cultivar 'Zhongmu No. 1′ was treated with 50 mM NaHCO 3 (0, 4, 8, 12 and 24 h) to study the resulting enzyme activity and changes in mRNA, miRNA and metabolites in the roots. The results showed that the enzyme activity changed significantly after alkali stress treatment. The genomic analysis revealed 14,970 differentially expressed mRNAs (DEMs), 53 differentially expressed miRNAs (DEMis), and 463 differentially accumulated metabolites (DAMs). Combined analysis of DEMs and DEMis revealed that 21 DEMis negatively regulated 42 DEMs. In addition, when combined with Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of DEMs and DAMs, we found that phenylpropanoid biosynthesis, flavonoid biosynthesis, starch and sucrose metabolism and plant hormone signal transduction played important roles in the alkali stress response. The results of this study further elucidated the regulatory mechanism underlying the plant response to alkali stress and provided valuable information for the breeding of new saline-alkaline tolerance plant varieties. [Display omitted] • Integrate transcriptome and metabolomics to analyse the alkaline stress of alfalfa. • Candidate genes, miRNAs and metabolites in response to alkaline stress in alfalfa were identified. • Alfalfa response to alkali is related to abscisic acid, ethylene, jasmonic acid and auxin signal transduction pathways. • Flavonoid synthesis was found to be associated with alkaline stress in alfalfa. • Five new miRNAs were identified, which could respond to alkaline stress. [ABSTRACT FROM AUTHOR]

Soil salinity constrains growth, development and yield of alfalfa (Medicago sativa L.). To illustrate the molecular mechanisms responsible for salt tolerance, a comparative proteome analysis was explored to characterize protein profiles of alfalfa seedling roots exposed to 100 and 200 mM NaCl for three weeks. There were 52 differentially expressed proteins identified, among which the mRNA expressions of 12 were verified by Real-Time-PCR analysis. The results showed increase in abundance of ascorbate peroxidase, POD, CBS protein and PR-10 in salt-stressed alfalfa, suggesting an effectively antioxidant and defense systems. Alfalfa enhanced protein quality control system to refold or degrade abnormal proteins induced by salt stress through upregulation of unfolded protein response (UPR) marker PDIs and molecular chaperones (eg. HSP70, TCP-1, and GroES) as well as the ubiquitin‐proteasome system (UPS) including ubiquitin ligase enzyme (E3) and proteasome subunits. Upregulation of proteins responsible for calcium signal transduction including calmodulin and annexin helped alfalfa adapt to salt stress. Specifically, annexin (MsANN2), a key Ca2+-binding protein, was selected for further characterization. The heterologous of the MsANN2 in Arabidopsis conferred salt tolerance. These results provide detailed information for salt-responsive root proteins and highlight the importance of MsANN2 in adapting to salt stress in alfalfa. • Comparative proteomics study revealed 52 salt-responsive proteins of alfalfa root. • Upregulation of the proteins involved in antioxidation and defense suggested amelioration of oxidative stress. • Alfalfa enhanced protein quality control system (UPR and UPS) to refold or degrade abnormal proteins induced by salt stress. • The heterologous of the MsANN2 in Arabidopsis conferred salt tolerance. [ABSTRACT FROM AUTHOR]

Gastrointestinal nematode (GIN) infection poses the most significant obstacle to the sustainable development of small ruminant (sheep and goat) farming globally. Resistance of GINs to synthetic anthelmintic drugs has led to rising interest in exploring alternative methods for parasite control, such as the utilization of bioactive plants with anti-parasitic properties. In this investigation, black seed (Nigella sativa), a shrub high in secondary antioxidant compounds, and sericea lespedeza (Lespedeza cuneata), a perennial legume high in tannins with anti-parasitic properties were combined to determine if two bioactive plants containing different types of secondary compounds can provide a stronger anti-parasitic effect than sericea lespedeza alone. In a 49-day trial, naturally parasitized 6–7-month-old intact male Spanish goats (n = 15/treatment) were fed pelletized feeds encompassing sericea lespedeza leaf meal (SL), a combination of black seed meal (BS) and sericea lespedeza leaf meal (BS-SL - 75 % SL, 25 % BS), or alfalfa (Medicago sativa , control parasitized; CONP), with an additional group of dewormed kids given the alfalfa pellets (Control treated; CONT). Weekly measurements of animal weights and samples of blood and feces were collected to determine the packed cell volume (PCV), GIN fecal egg counts (FEC), and coccidia fecal oocyte counts (FOC), respectively. All animals were processed at the end of the trial (60 total), with adult Haemonchus contortus worms recovered from the abomasum of each goat for counting and sex determination. Carcass weights were recorded after processing. Goats given the SL and BS-SL treatments had lower FEC (P<0.05) than the parasitized alfalfa (CONP) goats. At the end of the study, the SL and BS-SL groups' FOC values were lower (P < 0.05) than the CONT and CONP groups. A rise in PCV values was seen over time for all groups; SL, BS-SL, and CONT animals exhibited higher PCV values (P < 0.05) in comparison to the CONP goats. The parasitized goats fed SL-only pellets showed greater feed intake and animal body weights (P < 0.05) compared to goats fed BS-SL or alfalfa pellets. However, the treatments had no effect on the weight of the goats' carcasses. Although the H. contortus adult worm counts in the CONT goats (alfalfa-dewormed) were lower (P < 0.05) than in the CONP goats (alfalfa-parasitized), they did not differ from the SL or BS-SL animals. This study indicates that sericea lespedeza leaf meal pellet diet, either by itself or in combination with black seed meal, showed promising results as an anthelmintic and may prove to be an effective alternative to exclusive use of conventional deworming drugs. The addition of black seed did not appear to enhance the effectiveness of sericea lespedeza in this study. • Study evaluates natural alternatives to anthelmintics in small ruminants. • Experiment on goats' tests effects of sericea lespedeza and black seed meals. • Results: SL and BS-SL diets significantly reduce gastrointestinal nematode eggs. • Both treatments also lower coccidia counts, indicating better animal health. • SL diet increases feed intake and weight gain, boosting ruminant productivity. [ABSTRACT FROM AUTHOR]

Phosphorus is a nonrenewable source of fertilization, which will challenge the future of food production and cropland sustainability worldwide. Crop diversity is known to promote greater productivity, yet the capacity of crop diversity to alleviate productivity dependence on nonrenewable fertilization and promote multiple ecosystem services remains virtually unknown. Here, we conducted a field experiment to quantify the contribution of maize-alfalfa intercropping to soil biodiversity and multiple ecosystem services under contrasting phosphorus fertilization levels. Results showed that unfertilized intercropping can support higher levels of ecosystem services such as soil microbial habitat, plant-soil mutualism, nutrient cycling, and soil carbon storage compared to phosphorus-fertilized monoculture. Intercropping could optimize the delivery of soil diversity and multiple ecosystem services override phosphorus, including microbial diversity, weighted ecosystem services, productivity stability and sustainability, and soil microbial habitat were 5–30 times higher, respectively. Unfertilized intercropping also helped to reduce important tradeoffs between productivity and soil microbial diversity compared with fertilized monoculture. Together, our results provide evidence that intercropping can optimize crop use of phosphorus, and promote multiple important ecosystem services, which can help alleviate global dependence on nonrenewable, and often environmentally deleterious fertilizer inputs. [Display omitted] • Unfertilized intercropping support larger levels of multiple ecosystem services. • Intercropping reduce important tradeoffs in productivity and soil biodiversity. • Intercropping alleviates the dependence of ecosystem services on fertilization. • Intercropping optimized the delivery of multiple services and overridden phosphorus. [ABSTRACT FROM AUTHOR]

Establishing multi-species swards is a common objective in agricultural forage production systems around the world. Managing pasture productivity and sward resilience is especially challenging in semi-arid environments where mixtures of perennial and self-regenerating annual species are expected to co-exist while growing in competition under water-limited conditions. This review explores the implications of competition dynamics and the practice of changing row configuration at sowing as a prospective approach to managing the composition and productivity of multi-species swards. It uses three test case species representing different functional groups, including the perennial legume, lucerne (alfalfa; Medicago sativa L.), the perennial grass, phalaris (harding grass; Phalaris aquatica L.) and the self-regenerating annual legume, subterranean clover (Trifolium subterraneum L.). Early field studies showed consistent benefits of separating subterranean clover from other sward components at sowing, in terms of foliage dry matter and regeneration density of that species. However, total sward productivity either remained unchanged or declined compared to where all species were planted in the same drill row, especially where forage species were sown with grain cover crops. This is explained by a basic principle of ecology where dominant species drive total production in a short-term ecosystem such as a phased pasture. Constraining dominant species to fewer drill rows at sowing limited their production due to a transient restriction in resources, such as light. It is concluded that separating species in drill rows is a useful strategy for improving the abundance of transient or subordinate species when grown in mixtures with dominant species. However, to avoid perverse outcomes, care is required to ensure that the productivity of the dominant species is not compromised. Practical implications point towards maintaining the number of drill rows to which dominant species are sown. Further research is required to minimise drill row spacing, thus maximising the number of drill rows and increasing early plant coverage on a given area of land. The legacy effect of the pasture drill row is also highlighted, with most species remaining close to the original drill row for the life of a 3-year pasture phase in semi-arid environments. Increased concentration of plants led to enduring benefits in soil fertility, microbial diversity and abundance compared to the inter-row area, findings that warrant closer examination under a greater range of soil and climatic conditions. [Display omitted] • Separating species in different drill rows can improve species diversity in mixtures. • Biomass of transient species is improved when separated from dominant species. • Total biomass is reduced when dominant species are constrained to fewer drill rows. • Increased plant cover increases utilisation of light and other resources. • Reduced row spacing helps increase cover and abundance of dominant species [ABSTRACT FROM AUTHOR]

The present study was intended to evaluate the effect of forage source (alfalfa hay; ALF vs. corn silage; CS) along with a supplemental fat source (soybean oil; SO vs. rumen-inert palm fatty acids; PF) on growth performance, nutrient digestibility, and ruminal fermentation in dairy calves. Forty-eight new-born Holstein female calves (3 d old) were assigned to one of 4 treatments: (1) alfalfa hay with soybean oil (ALF–SO); (2) alfalfa hay with palm fatty acids (ALF–PF); (3) corn silage with soybean oil (CS–SO); (4) corn silage with palm fatty acids (CS–PF). Starter diets had equal amounts of forage (100 g/kg dry matter; DM) and fat source (30 g/kg DM). Calves were fed a constant amount of milk (d 1 to 63) and had ad libitum access to water and starters (d 1 to 83). The lowest and greatest starter intakes during the preweaning period occurred in ALF–SO and CS–PF, respectively. This coincided with forage × fat source interaction for average daily gain (ADG) during preweaning. The forage source affected total DM intake and ADG over the entire period, body weight (BW) at weaning, and final BW with greater values in calves that received CS compared with ALF. The concentrations of total short-chain fatty acids and butyrate were increased, whereas concentration of acetate and acetate:propionate ratio were decreased in the rumen of calves fed CS compared with ALF. Feeding CS increased urinary excretion of allantoin and, as a trend, total purine derivatives (PD) and estimated microbial protein synthesis in comparison with ALF. The fat source affected starter intake, ADG, and BW postweaning with the highest values in PF. The digestibility of neutral detergent fiber, crude protein and, as a trend, organic matter were higher in calves fed PF compared with SO. Calves fed PF had lower ruminal ammonia-N concentration and urinary N excretion and greater urinary excretion of allantoin and total PD. Calves receiving SO had a lower ruminal protozoa population. In conclusion, supplementing starter diets with CS and PF is superior to ALF and SO. Interaction of the positive effects of CS and PF on performance underlines that concurrent supplementation of CS with PF is especially recommendable in young calves before weaning. [ABSTRACT FROM AUTHOR]

Phosphite, a reduced form of phosphate, inhibits the growth and even has toxic effect on plants. To learn more about the mechanism of alfalfa responses to phosphite, the morphological and physiological characteristics, and the metabolites and transcript levels were comprehensively analyzed following the exposure of alfalfa seedlings to phosphite and phosphate under greenhouse conditions. The results showed that phosphite inhibited seedling growth and photosynthesis. However, the absorption efficiency of phosphite was higher than that of phosphate in roots, which was supported by increased total phosphorus concentration of 16.29% and 52.30% on days 8 and 12. Moreover, phosphite stress affected the synthesis of lipids and carbohydrates, which were reflected in enhanced glycolipid and sulfolipid in roots and amylose in shoots. Phosphite stress resulted in a decrease in indole acetic acid (IAA) in the whole plant and zeatin in the shoots, which could enable alfalfa to adapt to the phosphite environment. Some genes involved in phosphate starvation response included SPX , phosphate response regulator2 , and inorganic phosphate transporter 1–4 (PHT1;4) in roots were affected by phosphite stress. In addition, some genes that are involved in stress responses and DNA repair were induced by phosphite stress. These observations together suggest that alfalfa responds to phosphite stress by inhibiting growth, regulating the genes induced by phosphate starvation, improving oxidative protection, promoting DNA repair, and adjusting the IAA and zeatin signaling transductions. Our findings provide novel insights into the molecular response to phosphite stress in alfalfa. • Phosphite inhibited the growth of alfalfa seedlings. • Phosphite inhibited nitrogen assimilation and promoted sulfate assimilation. • Phosphite stress increased Phosphate starvation induced (PSI) genes. • Phosphite induced stress response genes and promoted DNA repair in alfalfa. [ABSTRACT FROM AUTHOR]

Dark septate endophytes (DSE) colonize plant roots extensively and increase host plant growth and nutrition. However, the development of DSE-produced metabolites as plant biostimulants has been largely ignored. DSE growth curves and extracellular metabolite components were analyzed and the growth-promoting effects of DSE extracellular metabolites on alfalfa (Medicago sativa L.) grown for 4, 8 12, 16 and 20 days were evaluated. The growth curve of the DSE strain Alternaria sp. shows days 0-8 in the growth phase, days 8-16 in the stable phase, and days 16-20 in the senescent phase. The extracellular metabolite components of DSE were significantly different at different growth stages. The biomass of alfalfa was increased significantly by DSE extracellular metabolites (P < 0.05). Biomass of alfalfa inoculated with DSE extracellular metabolites more than doubled after growth for 8 days and nutrient availability also increased significantly compared with the uninoculated control. Six DSE extracellular metabolites, calycosin 7-galactoside, 1-[(5-amino-5-carboxypentyl)amino]-1-deoxyfructose, N2-fructopyranosylarginine, 2-(4-methyl-5-thiazolyl)ethyl hexanoate, kenposide B, and medinoside E, were significantly positively correlated with alfalfa biomass (P < 0.01). This study combines the DSE extracellular metabolites with plant and soil traits to provide a theoretical basis for the use of DSE metabolites in the product development of plant biostimulants. • The time factor significantly affects the growth and metabolism of DSE. • The major components of DSE extracellular metabolites are amino acids and peptides. • The addition of DSE metabolites can promote alfalfa growth and nutrient uptake. • DSE metabolites promote plant growth and nutrient uptake through multiple pathways. [ABSTRACT FROM AUTHOR]

Accurate estimation of the hydrological features in the unsaturated zone is mandatory for the effective planning of irrigation strategies. Irrigation scheduling depends on crop and soil type as well as climatic characteristics and is usually empirically conducted. This paper simulates the water flow in order to model the soil water balance in three agricultural fields (maize, cotton, alfalfa) located in the River Strymonas basin using the HYDRUS-1D model. The model is fed with meteorological data, soil data and soil moisture measurements. After the calibration, through HYDRUS-1D's inverse solution, model results were used to evaluate the irrigation activities applied in the pilot application fields in terms of irrigation dose, irrigation interval and soil moisture variation for the cultivation period. In addition, in order to measure the efficiency of the irrigation method evaluated in this work, water productivities for all three fields were compared with productivities yielded from similar applications and experiments as well as precision irrigation experiments found around the world at similar climates with the one at Nigrita. [ABSTRACT FROM AUTHOR]

[Display omitted] • Genes related to TPH degradation and plant growth promotion were predicted. • Strain KB1 has TPH-degrading and growth-promoting properties under stress. • Strain KB1 promotes alfalfa growth and improves soil health. • Strain KB1 recruits TPH-degrading bacteria to accelerate TPH degradation. • Revealed microbial-phytoremediation mechanisms in contaminated soils. Despite the effectiveness of microbial-phytoremediation for remediating total petroleum hydrocarbons (TPH)-contaminated soil, the underlying mechanisms remain elusive. This study investigated the whole-genome and biological activity of Rhodococcus erythropolis KB1, revealing its plant growth promotion (PGP), TPH degradation, and stress resistance capabilities. Phytoremediation (using alfalfa) and plant-microbial remediation (using alfalfa and KB1) were employed to degrade TPH. The highest TPH degradation rate, reaching 95%, was observed with plant-microbial remediation. This is attributed to KB1′s ability to promote alfalfa growth, induce the release of signaling molecules to activate plant antioxidant enzymes, actively recruit TPH-degrading bacteria (e.g., Sphingomonas , Pseudomonas , C1-B045), and increase soil nitrogen and phosphorus levels, thereby accelerating TPH degradation by both plants and microorganisms. This study demonstrates that R. erythropolis KB1 holds great potential for enhancing the remediation of TPH-contaminated soil through its multifaceted mechanisms, particularly in plant-microbial remediation strategies, providing valuable theoretical support for the application of this technology. [ABSTRACT FROM AUTHOR]

Spectral analysis of meat combined with chemometric analysis has been identified as a promising tool for authenticating livestock-animal diets. The objectives of this study were (i) to determine whether the visible-NIR spectrum of perirenal adipose tissue (PAT) and caudal adipose tissue (CAT) can reliably discriminate lambs pasture-finished for different durations before slaughter, and (ii) to analyze the kinetics of appearance and stabilization of the visible-NIR spectrum-based pasture signature in PAT and CAT. Four groups of 50–55 lambs were used over three years: lambs finished on lucerne pasture for 0 (L0, concentrate-fed in stall), 21 (L21), 42 (L42) and 63 (L63) days before slaughter. Partial least squares discriminant analysis was applied on PAT or CAT visible-NIR spectra to discriminate the groups. No one adipose tissue reliably discriminated the four groups, with less than 62 % lambs correctly classified. However, visible-NIR spectroscopy was able to discriminate stall-fed (L0) from pasture-finished (L21 + L42 + L63) lambs, with an accuracy of 93.8 % and 87.5 % lambs correctly classified based on PAT and CAT spectra, respectively. The lucerne pasture fingerprint (or signature) on visible-NIR spectrum appeared between 0 and 42 days in more than 95 % of lambs. It stabilized between 42 and 63 days in CAT, but had not stabilized within the range of grazing durations pre-slaughter explored in PAT. Further research into shorter and longer pasture-finishing durations could help determine more precisely the time required for the pasture signature to appear and stabilize in animal tissues. • Fat visible-NIRS was used to authenticate pasture-finishing duration in lamb. • Fat spectrum failed to discriminate lambs from 4 durations of pasture finishing. • Fat spectrum correctly discriminated stall-fed lambs from alfalfa-finished lambs. • Pasture signature appeared in fat between 0 and 42 d grazing pre-slaughter. • Pasture signature has stabilized at different times depending on the tissue. [ABSTRACT FROM AUTHOR]

Intercropping alfalfa (Medicago sativa) with grass offers yields equal to or greater than alfalfa monoculture, improves the quality of silaged fodder, and enhances resilience to drought and other stresses. Inoculating either alfalfa monoculture or mixed cultures (with a festucoid hybrid (Festulolium pabulare)) with plant growth-promoting rhizobacteria (PGPR) could potentially enhance plant growth, yield and soil quality. A monoculture of alfalfa and three different mixed cultures of alfalfa and festucoid hybrid at ratios 1:1, 2:1, and 3:1 were sown at a seeding rate of 30 kg·ha-1 on small-scaled field plots (3 × 10 m). The soil type was Luvisol, either uninoculated or inoculated (⁓10.5 log10 CFUm-2) with a commercial PGPR consortium containing (Azotobacter , Sinorhizobium meliloti , Bacillus megatherium). At the end of the trial, mixed soil samples (comprising 8 probes to a depth of 10 cm) were collected, and their biological properties were determined. Mixed cultures of alfalfa with the festucoid hybrid decreased nitrification; urease was lower by 8.5 % (alfalfa:festucoid 1:1), 36.5 % (2:1), and 49.7 % (3:1) compared to alfalfa control. d -glucose-induced respiration was higher by 55.4 % (2:1) and by 23.1 % (3:1), along with a negative trend in the nitrifying Archaea abundance. Nitrososphaeria relative abundance decreased from 4.5 % (1:1) to 9.4 % (3:1) compared to the control. Inoculation indirectly affected nitrogen (N) turnover in the mixed variants by increasing urease (2:1 inoculated 49.1 % over 2:1 uninoculated; 3:1 inoculated 36.5 % over 3:1 uninoculated value) and increased the relative abundance of Nitrososphaeria (alfalfa inoculated 7.3 % and 2:1 inoculated 4.2 % over uninoculated control). Inoculation enhanced phosphatase activity (1:1 inoculated 11.4 %; 2:1 inoculated 21.8 %, 3:1 inoculated 16.2 % over respective uninoculated values), specific soil respiration (alfalfa inoculated 146 %, 1:1 inoculated 192 %, 2:1 inoculated 3 % over uninoculated values), and brought a positive tendency in copiotrophic (Actinobacteria) relative abundance (alfalfa inoculated 10.2 %, 1:1 inoculated 6.1 %, 3:1 inoculated 3.4 % over respective uninoculated values), albeit it decreased fungal biomass. An increased rate of N 2 fixation and N assimilation in variants with high alfalfa: grass ratio decreased nitrification and increased mineralization of specific substrates. Inoculation neither directly enriched the soil with the introduced PGPR taxa nor shifted soil microbial diversity. However, it did prompt tendencies in community composition changes towards a higher proportion of nitrifiers and copiotrophs. Despite multiple changes in the tested experimental variants, no significant effect on the plant biomass of harvested crops was observed during the three years. [Display omitted] [ABSTRACT FROM AUTHOR]

In the context of global food security and climate change, intercropping has been proposed as an effective agricultural practice to improve crop productivity and land use efficiency. The objectives of this study were to evaluate the total yield potential, land equivalent ratio, and economic benefits of one annual medic (Medicago polymorpha L.) and two alfalfa (M. sativa L.) cultivars intercropped with camelina [ Camelina sativa (L.) Crantz] at two different seeding rates (15 and 30 kg ha−1) in comparison with that of monocrops. Results showed that medic and alfalfa intercropped with camelina had greater land equivalent ratio (LER) value (∼1.63) and productivity than the monocropping systems. The intercropped camelina on average yielded (1045 kg ha−1 across the planting pattern) 17 % less than that of monocrop (1261 kg ha−1). Particularly, the seed yield of intercropped camelina + alfalfa 'XJDY' on average was 1295 kg ha−1 (across the planting pattern), exceeding the mean seed yield of 1261 kg ha−1 when it was monocropped, and produced the mean economic benefit of $3123 ha−1, which was about 1.3–1.9-fold greater than each of the species monocropped. Additionally, although the forage nutritional values of Medicago species or camelina seed oil quality varied among the different intercropping systems, they fell within the ranges reported in other studies. As alfalfa can be harvested multiple times during the growing season, we would like to highlight the potential for double-season (spring and fall) planting of camelina into the alfalfa 'XJDY' considering the shorter growing cycle of camelina (about three months from seeding to harvest). In summary, the alfalfa-camelina intercropping in this study showed great potential to improve land use efficiency, enhance productivity, and increase economic benefits, and thus can be used as a promising way to support the development of sustainable agriculture. [Display omitted] • Agronomic performance of Medicago-camelina intercropping was evaluated in eastern China. • Intercropping increased land use efficiency and productivity compared with monocropping. • Alfalfa 'XJDY'-camelina was the most profitable among tested intercropping patterns. • Intercropping showed no effect on forage nutritive value and camelina oil quality. • Camelina shows the great potential to be spring or fall-intercropped into alfalfa forage. [ABSTRACT FROM AUTHOR]

The annual dynamics of carbon dioxide (CO 2) fluxes for irrigated and rainfed alfalfa (Medicago sativa L.) in the Southern Great Plains of the United States of America (USA) under different watering regimes are not yet fully understood. The main objective of this study was to examine the dynamics of eddy covariance (EC) measured CO 2 fluxes in relation to various biophysical factors and hay harvests for irrigated and rainfed alfalfa in central Oklahoma, USA. The study also aimed to investigate the relationship between CO 2 fluxes and satellite-derived enhanced vegetation index (EVI) at different spatiotemporal scales and to assess the temporal variability in CO 2 fluxes and EVI to variable growing conditions and hay harvests. The cumulative hay yields were 7.15 t ha−1 (two harvests in 2019) in the rainfed field and ∼9 t ha−1 (4–5 harvests in 2020 and 2021) in the irrigated field. Having sufficient rainfall during April and May was crucial to achieve economically feasible yields of alfalfa during the first harvest in May. The availability of water strongly regulated the potential for regrowth and carbon uptake of alfalfa following harvesting. The alfalfa fields were near carbon neutral or a small carbon source from January to mid-March and carbon sink after the initiation of vegetative growth in mid-March. The alfalfa fields were strong carbon sinks (cumulative annual net ecosystem CO 2 exchange, NEE, up to −578 g C m−2 in irrigated field) on an annual scale. When accounting for the loss of carbon due to the removal of hay from the fields, the carbon balance of the alfalfa fields varied from small carbon sinks to small carbon sources, depending on the amount of hay harvested annually and the growing conditions. In general, the temporal patterns of CO 2 fluxes and EVI were similar in relation to growing conditions and hay harvests. However, some discrepancies and time lags were observed due to the coarse spatiotemporal resolution of the EVI products. Thus, it is essential to integrate two or more satellite products with different temporal and spatial resolutions to accurately monitor the frequent and varying sizes of hay harvests and vegetation regrowth after harvesting, and to simulate continuous time-series CO 2 fluxes. • Quantified annual dynamics of CO 2 fluxes from rainfed and irrigated alfalfa. • Sufficient April-May rainfall was vital for high yield at the first harvest in May. • Annual sums of net ecosystem CO 2 exchange reached up to −578 g C m−2. • CO 2 fluxes correlated strongly with finer resolution enhanced vegetation index. • Higher spatiotemporal images are needed for satellite-based modeling of CO 2 fluxes. [ABSTRACT FROM AUTHOR]

Researchers have investigated different techniques for synthesis of carbon dots. These techniques include Arc discharge, laser ablation, oxidation, water/solvothermal, and chemical vapor deposition. However, these techniques suffer from some limitations like the utilization of gaseous charged particles, high current, high temperature, potent oxidizing agents, non-environmentally friendly carbon sources, and the generation of uneven particle size. Therefore, there was a significant demand for the adoption of a new technology that combines the environmentally friendly aspects of both bio-based carbon sourcing and synthesis technique. Medicago sativa L (alfalfa)-derived N, S-CDs have been successfully synthesized via microwave irradiation. The N,S-CDs exhibit strong fluorescence (λ ex/em of 320/420 nm) with fluorescence quantum yield of 2.2 % and high-water solubility. The produced N,S-CDs were characterized using TEM, EDX, Zeta potential analysis, IR, UV–Visible, and fluorescence spectroscopy. The average diameter of the produced N, S-CDs was 4.01 ± 1.2 nm, and the Zeta potential was −24.5 ± 6.63 mv. The stability of the produced nano sensors was also confirmed over wide pH range, long time, and in presence of different ions. The synthesized N, S-CDs were employed to quantify the antibacterial drug, nifuroxazide (NFZ), by fluorescence quenching via inner filter effect mechanism. The method was linear with NFZ concentration ranging from 1.0 to 30.0 μM. LOD and LOQ were 0.16 and 0.49 μM, respectively. The method was applied to quantify NFZ in simulated gastric juice (SGJ) with % recovery 99.59 ± 1.4 in addition to pharmaceutical dosage forms with % recovery 98.75 ± 0.61 for Antinal Capsules® and 100.63 ± 1.54 for Antinal suspension®. The Method validation was performed in compliance with the criteria outlined by ICH. The suggested approach primarily centers on the first-time use of alfalfa, an ecologically sustainable source of dopped-CDs, and a cost-effective synthesis technique via microwave irradiation, which is characterized by low energy consumption, minimized reaction time, and the ability to control the size of the produced CDs. This is in line with the growing global recognition of the implementation of green analytical chemistry principles. [Display omitted] • Ultrafast synthesis of N,S-CDS via microwave treatment of alfalfa biomass. • Alfalfa was a sustainable, affordable, and natural precursor for N,S-CDS. • The N,S-CDS exhibit high stability, water solubility, and intense fluorescence. • The N,S-CDS were utilized as fluorescent sensors for nifuroxazide. [ABSTRACT FROM AUTHOR]

Feeding ruminants with high-quality forage can enhance digestibility and reduce methane production. Development of high-quality silage from leguminous plants with lactic acid bacteria can improve digestibility and it mitigate the greenhouse gas emissions. In this study, we developed a high-quality alfalfa silage with improved fermentation index and microbial dynamics using Levilactobacillus brevis -KCC-44 at low or high moisture (LM/HM) conditions and preserved it for 75 or 150 days. Alfalfa fermentation with L. brevis enhances acidification and fermentation characteristics primarily due to the dominance of lactic acid bacteria (LAB) L. brevis (>95%) compared to alfalfa fermented with epiphytic LAB. The inoculant L. brevis improved the anaerobic fermentation indexes resulting in a higher level of lactic acid in both high (10.0 ± 0.12 & 8.90 ± 0.31%DM) and low moisture (0.55 ± 0.08 & 0.39 ± 0.0 %DM) in 75 and 150 days respectively, compared to control silage. In addition, the marginal amount of acetic acid (range from 0.23 ± 0.07 to 2.04 ± 0.27 %DM) and a reduced level of butyric acid (range between 0.03 ± 0.0 to 0.13 ± 02 %DM) was noted in silage treated with LAB than the control. The LAB count and abundance of Levilactobacillus were higher in alfalfa silage fermented with L. brevis. Microbial richness and diversity were reduced in alfalfa silage treated with L. brevis which prompted lactic acid production at a higher level even for a prolonged period of time. Therefore, this L. brevis is an effective inoculant for producing high-quality alfalfa silage since it improves fermentation indexes and provides reproducible ensiling properties. [Display omitted] • High-quality forage can reduce methane production and improve digestibility. • Extended storage times for silage remain an issue due to aerobic instability. • Fermentation capability L. brevis was studied on alfalfa silages. • L.brevis heightened lactic acid primarily due to dominance of LAB. • Metagenomic revealed that L. brevis dominated in inoculated silages. [ABSTRACT FROM AUTHOR]

Nanoparticle (NP) pollution has negative impacts and is a major global environmental problem. However, the molecular response of alfalfa (Medicago sativa L.) to titanium dioxide nanoparticles (TiO 2 NPs) is limited. Herein, the dual effects of TiO 2 NPs (0–1000 mg L−1) on carbon (C) and nitrogen (N) metabolisms in alfalfa were investigated. The results showed that 500 mg L−1 TiO 2 NPs (Ti-500) had the highest phytotoxicity in the C/N metabolizing enzymes; and it significantly increased total soluble sugar, starch, sucrose, and sucrose‐phosphate synthase. Furthermore, obvious photosynthesis responses were found in alfalfa exposed to Ti-500. By contrast, 100 mg L−1 TiO 2 NPs (Ti-100) enhanced N metabolizing enzymes. RNA-seq analyses showed 4265 and 2121 differentially expressed genes (DEGs) in Ti-100 and Ti-500, respectively. A total of 904 and 844 differentially expressed proteins (DEPs) were identified in Ti-100 and Ti-500, respectively. Through the physiological, transcriptional, and proteomic analyses, the DEGs and DEPs related to C/N metabolism, photosynthesis, chlorophyll synthesis, starch and sucrose metabolism, and C fixation in photosynthetic organisms were observed. Overall, TiO 2 NPs at low doses improve photosynthesis and C/N regulation, but high doses can cause toxicity. It is valuable for the safe application of NPs in agriculture. [Display omitted] • The-affected metabolic pathways were carbon/nitrogen metabolism and photosynthesis. • Ti-500 resulted in phytotoxicity on photosynthesis in leaves. • TiO 2 NPs dose-dependently affected carbon and nitrogen metabolism in alfalfa. • TiO 2 NPs interfered with starch and sucrose metabolism and chlorophyll synthesis. [ABSTRACT FROM AUTHOR]

This study evaluates UVC light as an alternative to improve the microbiological quality of seeds intended for sprouting. UVC treatments with light exposure times up to 120 and 180 min were tested to reduce native bacterial population (NBP) in alfalfa, wheat, and chia seeds. The effects of the UVC light treatments on germination yield and rate, growth kinetics and antioxidant activity of sprouts were also evaluated. Results demonstrated that reductions of NBP after 180 min of UVC light exposure were dependent on the type of seed and their physical characteristics. The Weibull model demonstrated good fitness to experimental data (R2 >0.96) for the concave upward inactivation kinetics (n <1). Besides, the tested UVC light treatments did not affect the germination yield of the studied seeds and decreased the germination time to attain the commercial height of alfalfa and chia sprouts. The UVC light treatments applied on the studied seeds influenced positively the antioxidant properties when sprouting. The application of UVC light to alfalfa and wheat seeds destined for sprouting might be an alternative to improve their microbiological quality of seeds, without affecting their germination properties; while the nutritional benefits of its application, primordially focused to enhance its antioxidant properties of sprouts can be considered for chia and alfalfa seeds. • Effective inactivation of native microbiota of alfalfa and wheat seeds by UVC light. • UVC light treatments improves germination rate of alfalfa and chia seeds. • Antioxidant activity is enhanced by UVC light treatments of seeds when sprouting. [ABSTRACT FROM AUTHOR]

This study aimed to enrich corn and alfalfa silages with γ-aminobutyric acid (GABA) by utilizing high GABA-producing Lentilactobacillus buchneri. Eleven strains were screened and L. Buchneri YM9 was distinguished for its superiority in GABA production, and it was subsequently applied as an inoculant on whole-crop corn and alfalfa silage. The silage treatments were control (without inoculant), AH35 (non-GABA producing strain), YM9 (high-GABA producing strain), and 40788 (commercial GABA producing strain). The results revealed that in corn silage, pH significantly declined at the initial ensiling stage (3–7 days), with the control having the lowest pH after 90 days. The control also exhibited the highest lactic acid, while L. buchneri treatments had elevated acetic acid. Similar trends were observed in alfalfa silage, with 30 % dry matter (DM) showing lower pH and higher organic acids. YM9-inoculated corn silage had higher DM loss, reduced water-soluble carbohydrates (WSC), but increased crude protein (CP) content. YM9 and 40788 treatments in whole-crop corn silage had lower glutamate (Glu) content post-ensiling, signifying effective GABA production. YM9 treatment maintained stable and higher glutamate decarboxylase (GAD) activity, resulting in the highest GABA accumulation in corn silage (1.97 g/kg DM). Likewise, YM9 and 40788 demonstrated significantly higher GABA content in 30 % (7.6 and 6.51 g/kg DM) and 40 % (5.23 and 5.32 g/kg DM) DM alfalfa silage. Beyond enhancing fermentation and nutrient preservation, YM9 strain shows promise in enriching whole-crop corn and alfalfa with ample GABA concentration, potentially exerting anticipated biological functions when consumed by animals. • L. buchneri YM9 was selected for high GABA synthesis among the strains screened. • YM9 strain efficiently preserved the nutrients quality of corn and alfalfa silage. • The YM9 strain sufficiently enriched the silages with GABA during fermentation. [ABSTRACT FROM AUTHOR]

In the understanding of the molecular interaction between plants and their microbiome, a key point is to identify simplified models of the microbiome including relevant bacterial and fungal partners which could also be effective in plant growth promotion. Here, as proof-of-concept, we aim to identify the possible molecular interactions between symbiotic nitrogen-fixing rhizobia and soil fungi (Trichoderma spp.), hence shed light on synergistic roles rhizospheric fungi could have in the biology of symbiotic nitrogen fixation bacteria. We selected 4 strains of the model rhizobium Sinorhizobium meliloti and 4 Trichoderma species (T. velutinum , T. tomentosum , T. gamsii and T. harzianum). In an experimental scheme of 4 ×4 strains x species combinations, we investigated the rhizobia physiological and transcriptomic responses elicited by fungal spent media, as well as spent media effects on rhizobia-host legume plant (alfalfa, Medicago sativa L.) symbiosis. Fungal spent media had large effects on rhizobia, specific for each fungal species and rhizobial strains combination, indicating a generalized rhizobia genotype x fungal genotype interaction, including synergistic, neutral and antagonistic effects on alfalfa symbiotic phenotypes. Differential expression of a high number of genes was shown in rhizobia strains with up to 25% of total genes differentially expressed upon treatment of cultures with fungal spent media. Percentages over total genes and type of genes differentially expressed changed according to both fungal species and rhizobial strain. To support the hypothesis of a relevant rhizobia genotype x fungal genotype interaction, a nested Likelihood Ratio Test indicated that the model considering the fungus-rhizobium interaction explained 23.4% of differentially expressed genes. Our results provide insights into molecular interactions involving nitrogen-fixing rhizobia and rhizospheric fungi, highlighting the panoply of genes and genotypic interactions (fungus, rhizobium, host plant) which may concur to plant symbiosis. • The plant symbiotic bacterium Sinorhizobium meliloti is affected by the presence of the fungus Trichoderma. • Trichoderma modulates a large fraction of Sinorhizobium meliloti genes. • 30% of transcriptomic changes are specific to S. meliloti and Trichoderma strains combinations. • Specific S. meliloti - Trichoderma strains combinations can produce synergistic effects on the quality of symbiosis. [ABSTRACT FROM AUTHOR]

This study was carried out for 2 years (2020 and 2021) to elucidate the effects of water stress on yield and quality of alfalfa cultivars, to determine the water-yield and quality relations and the susceptibility of cultivars to water stress. Three different irrigation treatments (100%, 75% and 50% of full irrigation, I 100 , I 75 and I 50) and 12 different alfalfa cultivars (SHN, Osmanağa, Şahin 42, Minerva, Elçi, Riviera Vicentina, Magna 601, Magnum V, Prosementi, Ezzelina, Sunter, Nimet) were used in present experiments conducted in randomized-blocks split-plots design with 3 replications to investigate effects of irrigation levels on water, yield and quality parameters of alfalfa. Present findings revealed that water stress decreased crop water productivity, but increased irrigation water productivity of alfalfa cultivars. Herbage yields of the cultivars varied between 48.20 and 64.98 t/ha, hay yields between 11.42 and 14.67 t/ha and protein yields between 2.22 and 3.07 t/ha. Alfalfa was found to be sensitive to water stress in terms of herbage yield, but more tolerant in terms of hay and protein yields. Morphological characteristics (plant height, number of stems per plant, number of main stem branches and root crown diameter) decreased with increasing water stress levels. In terms of quality traits, while acid detergent fiber (ADF), neutral detergent fiber (NDF), crude fiber and crude ash ratios increased, crude protein ratios decreased with increasing irrigation water levels. It was concluded based on present findings that water stress resulted in different responses of different alfalfa cultivars and although it increased quality traits in general, it decreased herbage, hay and protein yields. Again, based on present findings, Magna 601 cultivar could be recommended for semi-arid regions. • The increase in water stress caused a decrease in the protein ratio while increasing the herbage yield. • Although the same amount of irrigation water was given in alfalfa cultivars, their yields changed significantly. • The increase in water stress also increased the digestibility of alfalfa. • The water productivity of alfalfa varied highly according to the cultivars. • While alfalfa was sensitive to drought in herbage yield, it was tolerant to water stress in hay and protein yield. [ABSTRACT FROM AUTHOR]