Effects of γ-polyglutamic acid supplementation on alfalfa growth and rhizosphere soil microorganisms in sandy soil.

This study aimed at exploring the effects of γ-polyglutamic acid on the growth of desert alfalfa and the soil microorganisms in the rhizosphere. The study examined the effects of varying concentrations of γ-polyglutamic acid (0%-CK, 2%-G1, 4%-G2, 6%-G3) on sandy soil, the research investigated its impact on the growth characteristics of alfalfa, nutrient content in the rhizosphere soil, and the composition of bacterial communities. The results indicated that there were no significant differences in soil organic matter, total nitrogen, total phosphorus, total potassium, and available phosphorus content among the G1, G2, and G3 treatments. Compared to CK, the soil nutrient content in the G2 treatment increased by 14.81-186.67%, showing the highest enhancement. In terms of alfalfa growth, the G2 treatment demonstrated the best performance, significantly increasing plant height, chlorophyll content, above-ground biomass, and underground biomass by 54.91-154.84%. Compared to the CK treatment, the number of OTUs (operational taxonomic units) in the G1, G2, and G3 treatments increased by 14.54%, 8.27%, and 6.84%, respectively. The application of γ-polyglutamic acid altered the composition and structure of the bacterial community, with Actinobacteriota, Proteobacteria, Chloroflexi, Acidobacteriota, and Gemmatimonadota accounting for 84.14-87.89% of the total bacterial community. The G2 treatment significantly enhanced the diversity and evenness of soil bacteria in the rhizosphere. Redundancy analysis revealed that organic matter, total nitrogen, total potassium, moisture content, and pH were the primary factors influencing the structure of bacterial phyla. At the genus level, moisture content emerged as the most influential factor on the bacterial community. Notably, moisture content exhibited a strong positive correlation with Acidobacteriota, which in turn was positively associated with indicators of alfalfa growth. In summary, the application of γ-polyglutamic acid at a 4% ratio has the potential for improving sandy soil quality, promoting plant growth, and regulating the rhizosphere microbial community.

Host plant preference of Lygus hesperus (Hemiptera: Miridae) in 4 field crops: potato, alfalfa, carrot, and pea.

The western tarnished plant bug, Lygus hesperus (Knight), has emerged as a pest of potatoes (Solanum tuberosum L.) in the Lower Columbia Basin of Oregon and Washington. This species is generally found infesting several other field-grown crops in the region; however, their host preference is poorly understood. Thus, greenhouse cage experiments were conducted to evaluate L. hesperus host preference by simultaneously presenting adults with 4 host plants: potato, alfalfa, Medicago sativa L., carrot, Daucus carota L., and pea, Pisum sativum L. In addition, an oviposition test was conducted. The results indicated that L. hesperus actively chose as a host and as an oviposition substrate among the 4 host plants. We found a significantly higher number of adults on alfalfa and potato plants over carrot or pea plants at 6 h, 24 h, and 48 h after adults were released into the cage. However, 96 h after release, more L. hesperus were found in alfalfa. In addition, female L. hesperus strongly preferred potato and alfalfa plants as an oviposition substrate over carrot and pea plants at 96 h after release.

Phytochemical Profile, Bioactive Properties, and Se Speciation of Se-Biofortified Red Radish (Raphanus sativus), Green Pea (Pisum sativum), and Alfalfa (Medicago sativa) Microgreens.

The impact of selenium (Se) enrichment on bioactive compounds and sugars and Se speciation was assessed on different microgreens (green pea, red radish, and alfalfa). Sodium selenite and sodium selenate at a total concentration of 20 µM (1:1) lead to a noticeable Se biofortification (40-90 mg Se kg-1 DW). In green pea and alfalfa, Se did not negatively impact phenolics and antioxidant capacity, while in red radish, a significant decrease was found. Regarding photosynthetic parameters, Se notably increased the level of chlorophylls and carotenoids in green pea, decreased chlorophyll levels in alfalfa, and had no effect on red radish. Se treatment significantly increased sugar levels in green pea and alfalfa but not in red radish. Red radish had the highest Se amino acid content (59%), followed by alfalfa (34%) and green pea (28%). These findings suggest that Se-biofortified microgreens have the potential as functional foods to improve Se intake in humans.

Evaluating the Antioxidant and Antidiabetic Properties of Medicago sativa and Solidago virgaurea Polyphenolic-Rich Extracts.

The present study evaluated the antioxidant and antidiabetic properties of Medicago sativa and Solidago virgaurea extracts enriched in polyphenolic compounds. The extracts were obtained by accelerated solvent extraction (ASE) and laser irradiation. Then, microfiltration was used for purification, followed by nanofiltration used to concentrate the two extracts. The obtained extracts were analyzed to determine their antioxidant activity using DPPH radical scavenging and reducing power methods. The antidiabetic properties have been investigated in vitro on a murine insulinoma cell line (ß-TC-6) by the inhibition of α-amylase and α-glucosidase. M. sativa obtained by laser irradiation and concentrated by nanofiltration showed the highest DPPH• scavenging (EC50 = 105.2 ± 1.1 µg/mL) and reducing power activities (EC50 = 40.98 ± 0.2 µg/mL). M. sativa extracts had higher inhibition on α-amylase (IC50 = 23.9 ± 1.2 µg/mL for concentrated extract obtained after ASE, and 26.8 ± 1.1), while S. virgaurea had the highest α-glucosidase inhibition (9.3 ± 0.9 µg/mL for concentrated extract obtained after ASE, and 8.6 ± 0.7 µg/mL for concentrated extract obtained after laser extraction). The obtained results after evaluating in vitro the antidiabetic activity showed that the treatment with M. sativa and S. virgaurea polyphenolic-rich extracts stimulated the insulin secretion of ß-TC-6 cells, both under normal conditions and under hyperglycemic conditions as well. This paper argues that M. sativa and S. virgaurea polyphenolic-rich extracts could be excellent natural sources with promising antidiabetic potential.

Pollen Diet Diversity does not Affect Gut Bacterial Communities or Melanization in a Social and Solitary Bee Species.

Pollinators face many stressors, including reduced floral diversity. A low-diversity diet can impair organisms' ability to cope with additional stressors, such as pathogens, by altering the gut microbiome and/or immune function, but these effects are understudied for most pollinators. We investigated the impact of pollen diet diversity on two ecologically and economically important generalist pollinators, the social bumble bee (Bombus impatiens) and the solitary alfalfa leafcutter bee (Megachile rotundata). We experimentally tested the effect of one-, two-, or three-species pollen diets on gut bacterial communities in both species, and the melanization immune response in B. impatiens. Pollen diets included dandelion (Taraxacum officinale), staghorn sumac (Rhus typhina), and hawthorn (Crataegus sp.) alone, each pair-wise combination, or a mix of all three species. We fed bees their diet for 7 days and then dissected out guts and sequenced 16S rRNA gene amplicons to characterize gut bacterial communities. To assess melanization in B. impatiens, we inserted microfilament implants into the bee abdomen and measured melanin deposition on the implant. We found that pollen diet did not influence gut bacterial communities in M. rotundata. In B. impatiens, pollen diet composition, but not diversity, affected gut bacterial richness in older, but not newly-emerged bees. Pollen diet did not affect the melanization response in B. impatiens. Our results suggest that even a monofloral, low-quality pollen diet such as dandelion can support diverse gut bacterial communities in captive-reared adults of these bee species. These findings shed light on the effects of reduced diet diversity on bee health.

Inflammatory mediators, oxidative stress and genetic disturbance in rheumatoid arthritis rats supported by alfalfa seeds metabolomic constituents via blocking interleukin-1receptor.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by aggressive cartilage and bone erosion. This work aimed to evaluate the metabolomic profile of Medicago sativa L. (MS) (alfalfa) seeds and explore its therapeutic impact against RA in rats. Arthritis was induced by complete Freund's adjuvant (CFA) and its severity was assessed by the arthritis index. Treatment with MS seeds butanol fraction and interlukin-1 receptor antagonist (IL-1RA) were evaluated through measuring interlukin-1 receptor (IL-1R) type 1 gene expression, interlukin-1 beta (IL-1ß), oxidative stress markers, C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), prostaglandin E2 (PGE2), caspase-3 (Cas-3), intracellular adhesion molecule-1 (ICAM-1), DNA fragmentation, and chromosomal damage. Total phenolics/ flavonoids content in the ethyl acetate, butanol fraction and crude extract of MS seeds were estimated. The major identified compounds were Quercetin, Trans-taxifolin, Gallic acid, 7,4'-Dihydroxyflavone, Cinnamic acid, Kudzusaponin SA4, Isorhamnetin 3-O-beta-D-2'',3'',4''-triacetylglucopyranoside, Apigenin, 5,7,4'-Trihydroxy-3'-methoxyflavone, Desmethylxanthohumol, Pantothenic acid, Soyasapogenol E, Malvidin, Helilandin B, Stigmasterol, and Wairol. Treatment with MS seeds butanol fraction and IL-1RA enhanced all the biochemical parameters and the histopathological features of the ankle joint. In conclusion, Trans-taxifolin was isolated for the first time from the genus Medicago. MS butanol fraction seeds extract and IL-1 RA were considered as anti-rheumatic agents.

Effect of Medicago sativa seed powder (Plus vitamin E vs. vitamin E alone) on semen analysis in men with idiopathic infertility: A double blind randomized clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Different parts of the alfalfa plant (Medicago sativa L.), especially its seeds, have been introduced as a semen quality enhancer in the folk medicine of different regions of Iran as well as in the traditional Persian medicine (PM) books. The seeds of this plant are also used in many combined medicines to treat male infertility in PM. AIM OF THE STUDY: This study was designed to investigate the effect of M. sativa seed powder plus vitamin E vs. vitamin E alone on semen analysis of men with idiopathic infertility. MATERIALS AND METHODS: In this randomized clinical trial, 60 participants were randomly divided into two groups and then received M. sativa seed powder (6 g daily) or placebo for three months, as add-on to vitamin E (100 U). Results of semen analysis (sperm total count, motility, and morphology as well as semen volume) of these two groups and their changes were compared before and after treatment as the primary outcome. The number of pregnancies after one month and probable side effects were also assessed. RESULTS: After 12 weeks, sperm total count changed from 36.2 ± 21.8 × 106/ml to 48.5 ± 19.1 × 106/ml in M. sativa and from 39.5 ± 23.5 × 106/ml to 41.2 ± 20.9 × 106/ml in placebo, percentage of normal morphology changed from 1.8 ± 0.8% to 2.6 ± 1.2 % in M. sativa and from 2.0 ± 0.9% to 2.6 ± 1.2% in placebo and percentage of motile sperm changed from 36.5 ± 11.8 % to 39.7 ± 12.0% in M. sativa and from 39.3 ± 10.1 % to 38.1 ± 12.1% in placebo. The improvements in M. sativa group are significantly better than placebo group (P = 0.00, 0.01 and 0.04, respectively). However, semen volume decreased in both groups, but its changes were not significant. The number of pregnancies was four in the intervention and zero in placebo group. One case with abdominal bloating and one with respiratory allergy withdrew from the intervention group. No other adverse effect was reported. CONCLUSIONS: In this study, significant improvement was detected in sperm parameters (except semen volume) of men with idiopathic infertility in M. sativa plus vitamin E group in compare to vitamin E alone after 12 weeks. These findings suggest that the M. sativa, as a complementary therapy, may have a beneficial effect on semen quality. More clinical trials with larger sample size are needed.

Selenite resistance and biotransformation to SeNPs in Sinorhizobium meliloti 1021 and the synthetic promotion on alfalfa growth.

Toxic selenite, commonly found in soil and water, can be transformed by microorganisms into selenium nanoparticles (SeNPs) as part of a detoxification process. In this study, a comprehensive investigation was conducted on the resistance and biotransformation of selenite in Sinorhizobium meliloti 1021 and the synergistic impact of SeNPs and the strain on alfalfa growth promotion was explored. Strain 1021 reduced 46% of 5 mM selenite into SeNPs within 72 h. The SeNPs, composed of proteins, lipids and polysaccharides, were primarily located outside rhizobial cells and had a tendency to aggregate. Under selenite stress, many genes participated in multidrug efflux, sulfur metabolism and redox processes were significantly upregulated. Of them, four genes, namely gmc, yedE, dsh3 and mfs, were firstly identified in strain 1021 that played crucial roles in selenite biotransformation and resistance. Biotoxic evaluations showed that selenite had toxic effects on roots and seedlings of alfalfa, while SeNPs exhibited antioxidant properties, promoted growth, and enhanced plant's tolerance to salt stress. Overall, our research provides novel insights into selenite biotransformation and resistance mechanisms in rhizobium and highlights the potential of SeNPs-rhizobium complex as biofertilizer to promote legume growth and salt tolerance.

Exploring root system architecture and anatomical variability in alfalfa (Medicago sativa L.) seedlings.

BACKGROUND: The growth of alfalfa (Medicago sativa L.) is significantly hampered by drought and nutrient deficiencies. The identification of root architectural and anatomical characteristics holds paramount importance for the development of alfalfa genotypes with enhanced adaptation to adverse environmental conditions. In this study, we employed a visual rhizobox system to investigate the variability in root system architecture (including root depth, root length, root tips number, etc.), anatomical features (such as cortical traits, total stele area, number and area of vessel, etc.), as well as nitrogen and phosphorus uptake across 53 alfalfa genotypes during the seedling stage. RESULTS: Out of the 42 traits measured, 21 root traits, along with nitrogen (N) and phosphorus (P) uptake, displayed higher coefficients of variation (CVs ≥ 0.25) among the tested genotypes. Local root morphological and anatomical traits exhibited more significant variation than global root traits. Twenty-three traits with CVs ≥ 0.25 constituted to six principal components (eigenvalues > 1), collectively accounting for 88.0% of the overall genotypic variation. Traits such as total root length, number of root tips, maximal root depth, and others exhibited positive correlations with shoot dry mass and root dry mass. Additionally, total stele area and xylem vessel area showed positive correlations with N and P uptake. CONCLUSIONS: These root traits, which have demonstrated associations with biomass and nutrient uptake, may be considered for the breeding of alfalfa genotypes that possess efficient resource absorption and increased adaptability to abiotic stress, following validation during the entire growth period in the field.

[Effect of Biochar with Phosphorus Fertilizer on Soil Nutrients, Enzyme Activity, and Nutrient Uptake of Alfalfa].

Biochar is widely used in agricultural production practices as a soil conditioner that can be used both alone and jointly with chemical fertilizer. However, there are few studies on the effects of the combined application of biochar and phosphate fertilizer with different particle sizes on soil and plants. In this experiment, pot experiments were used to study the effects of biochar with different particle sizes on soil nutrients, enzyme activity, and Alfalfa nutrient absorption under two phosphorus levels (according to diameter, the biochar was divided into C1:>1 mm and C2:<0.01 mm). This study showed that the combined application of biochar and phosphorus significantly improved soil nutrients, enzyme activity, and Alfalfa nutrient absorption. Among them, the C2 treatment significantly increased the soil available phosphorus content and phosphatase activity (P<0.05), whereas the C1 treatment had a significant effect on ammonium nitrogen, nitrate nitrogen, urease, and catalase activities (P<0.05). Moreover, the differences in nutrients and enzymes among biochar treatments with different particle sizes were affected by soil phosphorus levels. At the P0 level, there was no significant difference in ammonium nitrogen and nitrate nitrogen contents between the C1 and C2 treatments. At the P1 level, the contents of NH4+-N and NO3--N in the C1 treatment were 24.19% and 18.68% higher than those in the C2 treatment (P<0.05), but there was no significant difference between the C1 and C2 treatments. Phosphorus addition significantly increased the N and P contents of Alfalfa above ground and in the ground (P<0.05), but there was no significant effect on the nutrient content of Alfalfa between different particle sizes of biochar. In conclusion, biochar and phosphate fertilizer can be used as an effective means of soil improvement. In addition, when using biochar for soil improvement, the impact of particle size on soil nutrients and soil enzymes should be considered.

Differential ability of three bee species to move genes via pollen.

Since the release of genetically engineered (GE) crops, there has been increased concern about the introduction of GE genes into non-GE fields of a crop and their spread to feral or wild cross-compatible relatives. More recently, attention has been given to the differential impact of distinct pollinators on gene flow, with the goal of developing isolation distances associated with specific managed pollinators. To examine the differential impact of bee species on gene movement, we quantified the relationship between the probability of getting a GE seed in a pod, and the order in which a flower was visited, or the cumulative distance traveled by a bee in a foraging bout. We refer to these relationships as 'seed curves' and compare these seeds curves among three bee species. The experiments used Medicago sativa L. plants carrying three copies of the glyphosate resistance (GR) allele as pollen donors (M. sativa is a tetraploid), such that each pollen grain carried the GR allele, and conventional plants as pollen recipients. Different foraging metrics, including the number of GR seeds produced over a foraging bout, were also quantified and contrasted among bee species. The lowest number of GR seeds set per foraging bout, and the GR seeds set at the shortest distances, were produced following leafcutting bee visits. In contrast, GR seeds were found at the longest distances following bumble bee visits. Values for honey bees were intermediate. The ranking of bee species based on seed curves correlated well with field-based gene flow estimates. Thus, differential seed curves of bee species, which describe patterns of seed production within foraging bouts, translated into distinct abilities of bee species to move genes at a landscape level. Bee behavior at a local scale (foraging bout) helps predict gene flow and the spread of GE genes at the landscape scale.

Antioxidant status in the blood, liver, and muscle tissue of turkey hens receiving a diet with alfalfa protein concentrate.

This study aimed to investigate the impact of the oxidative potential of turkeys fed a diet with alfalfa protein concentrate (APC), used throughout the rearing period or periodically at 2-wk intervals. The research material consisted of 6-wk-old BIG 6 turkey hens kept in pens, 5 birds per pen in 6 replicates. The experimental factor was the addition of APC to the diet in the amount of 15 or 30 g/kg of diet. APC was administered in 2 ways: birds received a diet with APC throughout the experiment or periodically. In the latter case, the birds received the diet with APC for 2 wk, and then for 2 wk they received the standard diet without APC. Levels of nutrients in the diet; flavonoids, polyphenols, tannins, and saponins in APC; uric acid, creatinine, bilirubin, and some antioxidants in the blood; and enzyme parameters in the blood and tissues of turkeys were determined. The use of APC in the diet stimulated antioxidant processes, which could be seen in the values of the pro-oxidant-antioxidant parameters of the tissues and blood plasma of turkeys. The significant reduction in the H2O2 level (P = 0.042) and slight reduction in the MDA level (P = 0.083), accompanied by an increase in catalase (P = 0.046) activity in the turkeys continuously receiving APC in the amount of 30 g/kg of diet, as well as the increase in plasma antioxidant parameters (vitamin C, P = 0.042 and FRAP, P = 0.048) in these birds, reflects improvement in their antioxidant status. Thus continuous use of the APC supplement in the amount of 30 g/kg of diet proved to be a better feeding practice to optimize oxidative potential than periodic inclusion of APC.

Antagonistic impact on cadmium stress in alfalfa supplemented with nano-zinc oxide and biochar via upregulating metal detoxification.

Eco-toxicological estimation of cadmium induced damages by morpho-physiological and cellular response could be an insightful strategy to alleviate negative impact of Cd in agricultural crops. The current study revealed novel patterns of Cd-bioaccumulation and cellular mechanism opted by alfalfa to acquire Cd tolerance under various soil applied zinc oxide nanoparticles (nZnO) doses (0, 30, 60, 90 mg kg-1), combined with 2% biochar (BC). Herein, the potential impact of these soil amendments was justified by decreased Cd and increased Zn-bioaccumulation into roots by 38 % and 48 % and shoots by 51 % and 72 % respectively, with co-exposure of nZnO with BC. As, the transmission electron microscopy (TEM) and scanning electron microscopy and energy dispersive spectroscopy (SEM-EDS) ultrastructural observations confirmed that Cd-exposure induced stomatal closure, and caused damage to roots and leaves ultrastructure as compared to the control group. On the contrary, the damages to the above-mentioned traits were reversed by a higher nZnO dose, and the impact was further aggravated by adding BC along nZnO. Furthermore, higher nZnO and BC levels efficiently alleviated the Cd-mediated reductions in alfalfa biomass, antioxidant enzymatic response, and gaseous exchange traits than control. Overall, soil application of 90 mg kg-1 nZnO with BC (2 %) was impactful in averting Cd stress damages and ensuring better plant performance. Thereby, applying soil nZnO and BC emerge as promising green remediation techniques to enhance crop tolerance in Cd-polluted soil.

A 24-epibrassinolide treatment and intercropping willow with alfalfa increase the efficiency of the phytoremediation of cadmium-contaminated soil.

Cadmium contamination in agricultural soils threatens food security and human health, and that has caused widespread concern worldwide. Willow and alfalfa are widely used for the phytoremediation of cadmium (Cd)-contaminated soil, and willow NJU513 is the promising plant for remediating Cd-contaminated soil. In order to discuss the effect of intercropping willow NJU513 with alfalfa on the phytoremediation of Cd-contaminated soil, a pot-culture experiment was conducted in the greenhouse. The result showed that the phytoremediation of Cd-contaminated soil was enhanced by this intercropping because of the 25.90 % increase in the available Cd content. In order to increase the phytoremediation efficiency of Cd in the intercropping treatment, a 24-epibrassinolide (Brs) treatment was designed in the current study. The results showed that the phytoremediation of Cd-contaminated soil by willow and alfalfa improved following a Brs treatment because of the 16.32-74.15 % and 16.91-44.48 % increases in the plant biomass and available Cd content, respectively. Additionally, the extracted Cd by plants in the intercropping treatments with and without Brs was 0.56 and 0.31 mg pot-1, respectively. Transcriptome analyses of willow leaves revealed that Brs up-regulated the expression of genes related to calcium channel activity, calcium and zinc transmembrane transport, photosynthesis, catalase/antioxidant activity, glutathione metabolic processes and detoxification, phagosomes, and vacuoles, and that these upregulated genes promoted plant remediation efficiency and resistance to Cd stress. Brs promoted the phosphate ion transporter activity in willow leaves, which may have enhanced the solubilization of insoluble phosphate minerals by bacterial species (e.g., Vicinamibacterales, Bacillus, and Gaiella) to release Cd, ultimately leading to increased phytoremediation efficiency. In addition, plants with and without Brs treatments induced the bacteria-mediated transformation of available Cd to stable Cd. The study findings may be useful for improving the phytoremediation of Cd-contaminated paddy soil.

Converging alfalfa (Medicago sativa L.) and petroleum hydrocarbon acclimated ACC-deaminase containing bacteria for phytoremediation of petroleum hydrocarbon contaminated soil.

Plant assisted bioremediation of petroleum hydrocarbon contaminated soil is considered an effective green technology whereby accelerated degradation occurs due to converged effect of microorganisms and plants. However, survival and growth of microbes and plants under stress conditions is challenging task for success of the technology. In this study, plant growth promoting bacteria containing 1-aminocyclopropane-1-carboxylate (ACC)-deaminase activity and tolerance to petroleum hydrocarbon contamination were used in association with alfalfa for bioremediation of petroleum hydrocarbon contaminated soil. Eight pre-isolated bacterial isolates from soil having previous history of petroleum contamination were used in convergence with alfalfa on sand soil which was artificially contaminated (10 g crude oil per kg-1 of coarse textured soil). Combined effect of bacteria and plants on the degradation of petroleum hydrocarbons under controlled conditions of light and temperature was observed for a period of 60 days. The results of the study revealed that four bacterial isolates Bacillus subtilis strain PM32Y, Bacillus cereus strain WZ3S1, Bacillus sp. strain SM73 and Bacillus sp. strain WZ3S3 in association with alfalfa significantly degraded petroleum hydrocarbons. The most significant biodegradation (47%) of petroleum hydrocarbons was recorded in the experimental unit receiving PM32Y inoculation in association with alfalfa. Biodegradation of petroleum hydrocarbons was 33% with alone inoculation (without alfalfa) of PM32Y. The study revealed that combined use of bacteria and alfalfa plant is more efficient than alone application of either bacteria or plants for degradation of petroleum hydrocarbons.

This study provides the evidence for phytoremediation and significant degradation of petroleum hydrocarbons by using plant growth promoting bacteria (PGPB), containing 1-aminocyclopropane-1-carboxylate deaminase (ACC-deaminase) in association with alfalfa (Medicago sativa L.). The most significant biodegradation of petroleum hydrocarbons was recorded with a new combination of Bacillus subtilis strain PM32Y in association with alfalfa.

Integrated eco-physiological, biochemical, and molecular biological analyses of selenium fortification mechanism in alfalfa.

MAIN CONCLUSION: Foliar Se (IV) application at 100 mg/kg can act as a positive bio-stimulator of redox, photosynthesis, and nutrient metabolism in alfalfa via phenotypes, nutritional compositions, biochemistry, combined with transcriptome analysis. Selenium (Se) is an essential element for mammals, and plants are the primary source of dietary Se. However, Se usually has dual (beneficial/toxic) effects on the plant itself. Alfalfa (Medicago sativa L.) is one of the most important forage resources in the world due to its high nutritive value. In this study, we have investigated the effects of sodium selenite (Se (IV)) (0, 100, 200, 300, and 500 mg/kg) on eco-physiological, biochemical, and transcriptional mechanisms in alfalfa. The phenotypic and nutritional composition alterations revealed that lower Se (IV) (100 mg/kg) levels positively affected alfalfa; it enhanced the antioxidant activity, which may contribute to redox homeostasis and chloroplast function. At 100 mg/kg Se (IV) concentration, the H2O2, and malondialdehyde (MDA) contents decreased by 36.72% and 22.62%, respectively, whereas the activity of glutathione peroxidase (GPX) increased by 31.10%. Se supplementation at 100 mg/kg increased the plant pigments contents, the light-harvesting capacity of PSII (Fv/Fm) and PSI (ΔP700max), and the carbon fixation efficiency, which was demonstrated by enhanced photosynthesis (37.6%). Furthermore, alfalfa shifted carbon flux to protein synthesis to improve quality at 100 mg/kg of Se (IV) by upregulating carbohydrate and amino acid metabolic genes. On the contrary, at 500 mg/kg, Se (IV) became toxic. Higher Se (IV) disordered the plant antioxidant system, increasing H2O2 and MDA by 14.2 and 4.3%, respectively. Moreover, photosynthesis was inhibited by 20.2%, and more structural substances, such as lignin, were synthesized. These results strongly suggest that Se (IV) at a concentration of 100 mg/kg act as the positive bio-stimulator of redox metabolism, photosynthesis, and nutrient in alfalfa.

[Soil nematode community characteristics of alfalfa field with different growing ages in the semi-arid Loess Plateau of Central Gansu, Northwest China]. / 陇中黄土高原半干旱区不同种植年限紫花苜蓿土壤线虫群落分布特征.

To clarify the impacts of long-term alfalfa plantation on the soil nematode community, soil samples were collected from different alfalfa growing ages (2 a, 9 a, 18 a) in the semi-arid area of Loess Plateau in Central Gansu by Illumina Miseq sequencing technology. The main controlling factors affecting its community change were also explored. The results showed that soil nematode belongs to 2 classes, 7 orders, 16 families and 21 genera. Among them, Chromadorea was the dominant group (44.6%-81.4%), the relative abundance of which decreased with alfalfa growing ages. Paratylenchus, Helicotylenchus, Xiphinema, Pristionchus, Ditylenchus, Panagrolaimus, Longidorus, Aprutides, Isolaimium and Aglenchus were the special nematode species of alfalfa, among which Paratylenchus (54.1%), Helicotylenchus (23.9%) and Xiphinema (21.9%) were the dominant nematodes in 2 a, 9 a and 18 a alfalfa soil respectively. Plant-parasitic nematode was the dominant group in alfalfa soil (31.8%-67.1%), and its relative abundance decreased at first and then increased with alfalfa growing ages. Results of redundancy analysis showed that soil available phosphorus and total nitrogen were the dominant environmental factors affecting community structure of soil nematodes in the region.

Growth performance, nutrient digestibility, and ruminal fermentation of dairy calves fed starter diets with alfalfa hay versus corn silage as forage and soybean oil versus palm fatty acids as fat source.

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.

Effect of alfalfa on subsurface (tile) nitrogen and phosphorus loss in Ohio, USA.

Growing annual crops such as corn (Zea mays L.) can lead to considerable nutrient losses through subsurface drainage in agricultural fields, posing a serious threat to surface water quality in the midwestern United States. Perennial crops have the potential to reduce these nutrient losses. However, more comprehensive data are needed on the nutrient loss effect of perennial crops. We examined the effect of alfalfa (Medicago sativa L.) on nitrate-nitrogen (NO3 - -N), total nitrogen (TN), dissolved reactive phosphorus (DRP), and total phosphorus (TP) in subsurface drainage using a before-after-control-impact (BACI) experimental design with one control field (with annual crops) and one impact field (with alfalfa) each on two farms (Sites A and B) located in northwestern Ohio. The "Before" period (prior to planting alfalfa at the impact field) extended for 4 yr (2013-2017) at Site A and 6 yr (2011-2017) at Site B; the "After" period extended for an additional 2 yr at both sites. Reductions in the mean monthly discharge and loads of NO3 - -N, TN, DRP, and TP were significant at Site A, whereas the only significant change at site B was a reduction in the mean monthly TP load. Significant reductions in NO3 - -N loads were observed during spring and winter at Site A. In addition, alfalfa reduced the variability of discharge and nutrient loads through subsurface drainage at both sites. Our findings suggest that introducing alfalfa into annual crop rotations has the potential to reduce subsurface nutrient loads and increase the resiliency of agricultural systems.

Long-term alfalfa (Medicago sativa L.) establishment could alleviate phosphorus limitation induced by nitrogen deposition in the carbonate soil.

Phosphorus (P) limitation is a widespread problem of primary production in dryland submitted to persistent nitrogen (N) deposition. The legume alfalfa (Medicago sativa L.), which can fix N2, might potentially strengthen P limitation in dryland ecosystems and is widely distributed as forage. However, there is still unclear how alfalfa grassland mobilizes the soil P to meet its demand. In this experiment, alfalfa introduction was used for long-term revegetation to evaluate the P uptake of plants from deep soil and assess the P limitation induced by N deposition compared with fallow. Our results showed that alfalfa introduction increased the soil P storage significantly at 0-2.4 m soil depth (+0.74 Mg ha-1), whereas it decreased at 2.4-4.8 m soil depth (-0.21 Mg ha-1) after 15-year establishment. Alfalfa establishment increased soil organic P concentration (180.9 mg kg-1 vs. 67.2 mg kg-1) and its relative contribution to total P (19.64% vs. 8.08%) at 0-4.8 m. Alfalfa establishment also increased the concentration and proportion of labile and intermediate P fractions at 0-4.8 m (9.12 mg kg-1 vs. 6.87 mg kg-1, 1.12% vs. 0.98%; 16.06 mg kg-1 vs. 8.39 mg kg-1, 1.69% vs. 1.17%). Alfalfa introduction decreased the concentrated HCl-Pi (250.66 mg kg-1 vs. 229.32 mg kg-1, 36.81% vs. 28.91%) in 2.4-4.8 m soil depth. These results indicated that the deep root system of alfalfa grassland could promote the P mobilization from deep to shallow soil. The concentrated HCl-Pi may be the main potential P source of alfalfa from 2.4-4.8 m to 0-2.4 m of soil depth, and long-term establishment of alfalfa can alleviate P limitation caused by N deposition in carbonate soil. Our results suggested that species with deep roots (such as alfalfa) could be selected as an economical way to mitigate nitrogen deposition in drylands.