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Spring regrowth is an important trait for perennial plants including alfalfa, the most cultivated forage legume worldwide. However, the genetic and genomic basis of the trait is largely unknown in alfalfa due to its complex genetic background of the tetroploid genome. The objective of this study was to identify quantitative trait loci (QTLs) associated with spring regrowth using high-resolution genetic linkage maps we constructed previously. In total, 36 significant additive effect QTLs for the trait were detected. Among them, 10 QTLs individually explained more than 10% of the phenotypic variation (PVE) with four in P1 and six in P2. Six overlapped QTLs intervals were detected with two and four intervals distributed in P1 and P2, respectively. In P1, both overlapped genomic regions were located on homolog 7D. In P2, the four QTLs with PVE>10% were co-localized on homolog 6D. Meanwhile, six pairs of significant epistatic QTLs were identified in P2. Screening of potential candidate genes associated with four overlapped QTLs (qCP2019-8, qLF2019-5, qLF2020-4, and qBLUP-3) narrowed down one candidate annotated as MAIL1. The Arabidopsis homolog gene has been reported to play an important role in plant growth. Therefore, the detected QTLs are valuable resources for genetic improvement of alfalfa spring vigor using marker-assisted selection (MAS), and further identification of the associated genes would provide insights into genetic control of spring regrowth in alfalfa.

Medicago sativa L. (alfalfa), a leguminous crop, is vital globally for forage, crop rotation, and soil fertility. Though, the microbiome of alfalfa grown in uncultivated land has not been studied yet. Here, we employed the high-throughput 16S rRNA and nifH gene amplicon sequencing to unravel the rhizosphere microbiome and nodules endosymbionts of alfalfa grown in poor lawn and coastal sandy soils. The root nodule of alfalfa of coastal sands and poor lawn soil, Sinorhizobia/Ensifer accounted for 80.599% and 67.727% of the total microbiome, respectively. The class Actinobacteria (except Frankiales) and order Rhizobiales, Sphingomonadales, Burkholderiales, Pseudomonas, and Cytophagia were found significantly enriched. Culturable studies displayed the dominance of Bacillus followed by Arthrobacter, Microbacterium, and Streptomyces. Hydroponics studies revealed the efficiency of cultured isolates for nodulation and plant growth attributes. Overall studies decipher the soil indigenous microbiome associated with root nodule and rhizosphere of Alfalfa grown in uncultivated coastal and sandy land.

In Turkestan region of South Kazakhstan has large areas of fertile sierozem soils that are important for crop production and its accompanying economic development. And also, alfalfa (Medicago sativa) is an important forage crop grown for seed production. The soils are fertile loams, but because of the regions dry environment, they need to be irrigated. Field experiments were conducted from 2017 to 2020 on one-year or older alfalfa stands grown for seed production at various plots on sierozem soils in rainfed areas of Turkestan region to determine the influence of agricultural practices such as fertilizer use, pesticide use and conventional tillage on seed yield and dry hay of alfalfa stands. The findings of field research experiments indicated that agricultural practices of fertilizer use, pesticide use and conventional tillage was essential to obtain maximum seed yield and dry hay of alfalfa. But, it was determined that the best outputs tended to be obtained with Fertilizer use + Pesticide use + Conventional tillage.

Fifteen alfalfa varieties cultivated in Turkey were researched to compare the antioxidant potential of their seeds and find differences between the varieties. The total phenolic (TP) content, DPPH•, and ABTS•+ scavenging activities, and ferric-reducing antioxidant power (FRAP) of alfalfa seed methanol extracts were determined. TP contents of extracts were found between 37.03 and 54.04 mg GAE/g. ABTS assay results ranged between 0.100 and 0.158 mmol Trolox/g extract, and FRAP changed from 389.90 to 791.02 µmol Fe2+/g extract. The results demonstrated that the differences between the alfalfa varieties were significant (p < 0.01) in terms of analyzed characters. © 2021, Centenary University. All rights reserved.

Background The root system architecture (RSA) of alfalfa (Medicago sativa L.) affects biomass production by influencing water and nutrient uptake, including nitrogen fixation. Further, roots are important for storing carbohydrates that are needed for regrowth in spring and after each harvest. Previous selection for a greater number of branched and fibrous roots significantly increased alfalfa biomass yield. However, phenotyping root systems of mature alfalfa plant is labor-intensive, time-consuming, and subject to environmental variability and human error. High-throughput and detailed phenotyping methods are needed to accelerate the development of alfalfa germplasm with distinct RSAs adapted to specific environmental conditions and for enhancing productivity in elite germplasm. In this study methods were developed for phenotyping 14-day-old alfalfa seedlings to identify measurable root traits that are highly heritable and can differentiate plants with either a branched or a tap rooted phenotype. Plants were grown in a soil-free mixture under controlled conditions, then the root systems were imaged with a flatbed scanner and measured using WinRhizo software. Results The branched root plants had a significantly greater number of tertiary roots and significantly longer tertiary roots relative to the tap rooted plants. Additionally, the branch rooted population had significantly more secondary roots > 2.5 cm relative to the tap rooted population. These two parameters distinguishing phenotypes were confirmed using two machine learning algorithms, Random Forest and Gradient Boosting Machines. Plants selected as seedlings for the branch rooted or tap rooted phenotypes were used in crossing blocks that resulted in a genetic gain of 10%, consistent with the previous selection strategy that utilized manual root scoring to phenotype 22-week-old-plants. Heritability analysis of various root architecture parameters from selected seedlings showed tertiary root length and number are highly heritable with values of 0.74 and 0.79, respectively. Conclusions The results show that seedling root phenotyping is a reliable tool that can be used for alfalfa germplasm selection and breeding. Phenotypic selection of RSA in seedlings reduced time for selection by 20 weeks, significantly accelerating the breeding cycle.

Background and Aim: Feed plays the most important role in supporting livestock productivity. There is a need for studies on the nutrient levels in feed absorbed by the body of livestock. The aim of this study was to investigate the effects of alfalfa (Medicago sativa L.) supplementation in feed on growth, small intestinal histomorphology, and digestibility in hybrid ducks. Materials and Methods: The study was conducted in vivo using 75 hybrid ducks, with three treatments and five replications. Each replication consisted of five ducks. The treatments were: T0=basal ration without any alfalfa supplementation; T1=basal ration+3% fresh alfalfa supplementation; and T2=basal ration+6% fresh alfalfa supplementation. Fresh alfalfa was quantified on the basis of dry matter content. Drinking water was provided ad libitum. The observed variables were growth performance, measured in terms of feed consumption, body weight gain, and feed conversion ratio; duodenum histomorphology, measured in terms of villus height, villus width, and crypt depth; digestibility, measured in terms of dry matter digestibility; and organic matter, crude protein, and crude fiber concentrations. The data were analyzed using variance analysis with a completely randomized design of one-way pattern with Statistical Product for Service Solution application of version 22. The data with significant differences were further analyzed using Duncan's new multiple range rest. Results: The results of the study showed that 3% alfalfa supplementation increased feed consumption and body weight gain of the hybrid ducks at 35 days of age compared with 0% and 6% supplementation. Furthermore, 3% alfalfa supplementation presented the best result in terms of villus height and duodenal crypt depth. Meanwhile, 6% alfalfa supplementation decreased nutrient digestibility in the ducks. Conclusion: These findings show that supplementation of feed with fresh alfalfa could have a significant effect on hybrid ducks in terms of growth performance, small intestinal histomorphology, and digestibility.

Endogenous viral elements (EVEs) have been for the most part described in animals and to a less extent in plants. The endogenization was proposed to contribute toward evolution of living organisms via horizontal gene transfer of novel genetic material and resultant genetic diversity. During the last two decades, several full-length and fragmented EVEs of pararetroviral and non-retroviral nature have been identified in different plant genomes, both monocots and eudicots. Prior to this work, no EVEs have been reported in alfalfa (Medicago sativa L.), the most cultivated forage legume in the world. In this study, taking advantage of the most recent developments in the field of alfalfa research, we have assessed alfalfa genome on the presence of viral-related sequences. Our analysis revealed segmented EVEs resembling two dsDNA reverse-transcribing virus species: Soybean chlorotic mottle virus (family Caulimoviridae, genus Soymovirus) and Figwort mosaic virus (family Caulimoviridae, genus Caulimovirus). The EVEs appear to be stable constituents of the host genome and in that capacity could potentially acquire functional roles in alfalfa’s development and response to environmental stresses.

To explore the effects of mowing frequency, fertilization, and their interaction on the yield and quality of alfalfa(Medicago sativa) in light saline-sodic soils, a field experiment with a split-plot design was conducted in the western Songnen Plain. The mowing frequency treatment plot(once, twice, three, and four times per year) was used as the main plot and the fertilization treatment plot(CK, N, P, and NPK) was the secondary plot. The results showed that the hay yield of alfalfa increased significantly(PPP+/Na+ increased significantly(PP2+ increased first and then decreased; and the ratio of Ca2+/Na+ tended to increase first and then stabilize. Application of NPK compound fertilizer significantly increased the content of crude protein and content of crude fat of alfalfa and decreased the content of crude fiber(PP

The current paper has presented the estimation results of ecological adaptability of the alfalfa samples. The purpose of the work was to assess the productivity and quality of green mass of the alfalfa samples from the IPI plant genetic resources gene bank and to identify the most adaptive ones according to the trait ‘green mass productivity’ and ‘raw protein percentage’. The study of the collection alfalfa samples was carried out in the southern part of the Rostov region on the plots of the “ARC “Donskoy” in the breeding crop rotation of perennial grasses in 2016–2018. The objects of study were 30 alfalfa samples from the collection of N.I. Vavilov IPI from different countries (Canada, the USA, Peru, France). The variety ‘Rostovskaya 90’ was used as a standard one. The estimation of alfalfa samples on the presence of adaptive properties in them according to the trait ‘green mass productivity’ showed that the most valuable samples in present practical breeding work are the samples ‘K-32873’, ‘K-33299’, ‘K-42684’, ‘K-42249’, ‘K-78803’ with weak responsiveness to changes in environmental conditions; the samples ‘K-36104’, ‘K-48778’, ‘K-42694’, ‘K-45715’, ‘K-47800’, ‘K-47802’, ‘K-43260’ with high resistance to stress; the samples ‘K-43272’, ‘K-50545’, ‘K-47806’, ‘K-47807’ with genetically flexible genotypes. When breeding according to the trait ‘raw protein percentage’, the samples ‘K-47807’, ‘K-47804’, ‘K-42712’ possessing a high raw protein percentage and resistance to changes in this trait are important for further work.

The aim of this investigation was to determine appropriate perennial grasses and their most suitable mixture ratios, which can be grown with alfalfa in binary mixtures. The research was conducted during 2009-2011 in Isparta (37.50 °N, 30.32 °E), located in the Western Mediterranean climate conditions. In the study, alfalfa was planted as a binary mixture with perennial ryegrass (Lolium perenne L.), orchardgrass (Dactylis glomerata L.), meadow fescue (Festuca pratensis Huds.), and smooth bromegrass (Bromus inermis Leyss.) in a randomized block design with three replications. Sowing ratios of alfalfa in the mixtures varied 20, 30, and 40%, and grass species ratios were 80, 70, and 60%. In the study, significant differences were found among the applications regarding hay yield, crude protein content, crude protein yield, NDF, ADF, land equivalent ratio (LER), and grass ratio in the hay (GR). The highest hay yield was obtained from alfalfa + orchardgrass and alfalfa + smooth bromegrass binary mixtures (25.98 and 25.78 t ha-1, respectively). Alfalfa + perennial ryegrass and alfalfa + meadow fescue mixtures gave the highest crude protein contents as 14.93 and 14.80%, respectively. The highest LER values were observed on perennial ryegrass and smooth bromegrass binary mixtures, and the highest grass ratios were determined in orchardgrass and smooth bromegrass mixtures made with alfalfa. Increasing the alfalfa sowing ratio in the mixtures positively affected all quality characteristics such as crude protein, NDF, and ADF. It was concluded that orchardgrass and smooth bromegrass binary mixtures with alfalfa produced high yield and quality hay and that the mixture should contain 40% alfalfa and 60% grass.

Ramie (Boehmeria nivea) is noted for the production of a large biomass that has a high protein content and is rich in antioxidants. It may thus serve as a high-quality forage material to replace alfalfa and improve the meat quality of farmed animals. In this study, we evaluated the carcass characteristics and meat quality of goats when 0, 35%, 75%, and 100% of dietary alfalfa was replaced with ramie. Crude protein content (linear, P

Background Iron (Fe) is an essential plant nutrient. Its deficiency is a major constraint in crop production systems, affecting crop yield and quality. It is therefore important to elucidate the responses and adaptive mechanisms underlying Fe-deficiency symptoms in alfalfa. Materials and methods The experiment was carried out on 12-day-old alfalfa plants grown in hydroponics under Fe-sufficient and Fe-deficient conditions. Results The Fe-starved alfalfa showed decreased plant biomass, chlorophyll score, PSII efficiency, and photosynthesis performance index in young leaves under low Fe. Further, Fe shortage reduced the Fe, Zn, S and Ca concentration in root and shoot of alfalfa accompanied by the marked decrease of MsIRT1, MsZIP, MsSULTR1;1, MsSULTR1;2 and MsSULTR1;3 transcripts in root and shoot. It indicates that retardation caused by Fe-deficiency was also associated with the status of other elements, especially the reduced Fe and S may be coordinately attributed to the photosynthetic damages in Fe-deficient alfalfa. The ferric chelate reductase activity accompanied by the expression of MsFRO1 in roots showed no substantial changes, indicating the possible involvement of this Strategy I response in Fe-deficient alfalfa. However, the proton extrusion and expression of MsHAI1 were significantly induced following Fe-deficiency. In silico analysis further suggested their subcellular localization in the plasma membrane. Also, the interactome map suggested the partnership of MsFRO1 with plasma membrane H+-ATPase, transcription factor bHLH47, and nitrate reductase genes, while MsHAI1 partners include ferric reductase-like transmembrane component, plasma membrane ATPase, vacuolar-type H-pyrophosphatase, and general regulatory factor 2. In this study, SOD and APX enzymes showed a substantial increase in roots but unable to restore the oxidative damages in Fe-starved alfalfa. Conclusion These findings promote further studies for the improvement of Fe-starved alfalfa or legumes through breeding or transgenic approaches. Graphic Abstract

For both New Zealand and China, agriculture is integral to the economy, supporting primary production in both intensive and extensive farming systems. Grasslands have important ecosystem and biodiversity functions, as well providing valuable grazing for livestock. However, production and persistence of grassland and forage species (e.g. alfalfa) is not only compromised by overgrazing, climate change and habitat fragmentation, but from a range of pests and diseases, which impose considerable costs on growers in lost production and income. Some of these pest species are native, but increasingly, international trade is seeing the rapid spread of exotic and invasive species. New Zealand and China are major trading partners with significant tourist flow between the two countries. This overview examines the importance of grasslands and alfalfa in both countries, the current knowledge on the associated insect pest complex and biocontrol options. Identifying similarities and contrasts in biology and impacts along with some prediction on the impact of invasive insect species, especially under climate change, are possible. However, it is suggested that coordinated longitudinal ecological research, carried out in both countries using sentinel grass and forage species, is critical to addressing gaps in our knowledge of biology and impact of potential pests, along with identifying opportunities for control, particularly using plant resistance or biological control.

Alfalfa is an unconventional alternative legume and its potentiality for utilization can be improved by processing. In this study, alfalfa seeds were processed by different thermal and non-thermal techniques and evaluated for their physical properties, colour characteristics, nutritional composition, pasting profile and protein secondary structure. The results were analyzed using Principal Component Analysis to elucidate the effect of processing. Colour of the flour was more affected by thermal processing in comparison to non-thermal processing. Nutritional composition of alfalfa flour was significantly affected by germination as compared to other methods and germinated flour showed an increase in the protein content. All the processing treatments resulted in improving the pasting properties except for germination. Particularly, marked improvement in the pasting properties was observed by wet heat processing and soaking. ATR-FTIR exhibited change in the secondary structure of the protein as a result of processing and showed the dominance of intermolecular β-sheets in extrusion, germination and microwave treated flour and formation of anti-parallel β-sheets after dry heat processing. Wet heat processing and soaking treatment can be employed for alfalfa for enhancing its techno-functionality.

The legume plant alfalfa (Medicago sativa L.) is a widely cultivated perennial forage due to its high protein content, palatability, and strong adaptability to diverse agro-ecological zones. Alfalfa is a self-incompatible cross-pollinated autotetraploid species with tetrasomic inheritance. Therefore, maintaining excellent traits through seed reproduction is a prime challenge in alfalfa. However, the cutting propagation technology could enable consistent multiplication of quality plants that are genetically identical to the parent plant. The current study aimed to develop a simple, cost-effective, reproducible, and efficient hydroponic cutting method to preserve alfalfa plants and for molecular research. In this study, alfalfa landrace ‘Wudi’ was grown in hydroponics for 30 days and used as source material for cuttings. The top, middle and bottom sections of its stem were used as cuttings. The rooting rate, root length, and stem height of the different stem sections were compared to determine the best segment for alfalfa propagation in four nutrient treatments (HM, HM + 1/500H, HM + 1/1000H and d HM + 1/2000H). After 21 days of culture, the rooting rates of all the three stem types under four cutting nutrient solutions were above 78%. The rooting rate of the middle and bottom parts in HM + 1/1000 H and HM + 1/2000 H nutrient solutions reached more than 93%, with a higher health survey score (> 4.70). In conclusion, this study developed a de novo cutting propagation method that can be used to conserve and propagate germplasm in breeding programs and research. This method is a new report on the cutting propagation of alfalfa by hydroponics, which could supplement the existing cutting propagation methods.

Background The objective of this study was to evaluate the chemical compositions and microbial communities of salt-tolerant alfalfa silage. Salt-tolerant alfalfa was ensiled with no additive control, and cellulase for 30 and 60 to 90 days. In this study, the dry matter (DM) content of the raw material was 29.9% DM, and the crude protein (CP) content of the alfalfa was 21.9% CP. Results After 30 days of fermentation, the DM content with the cellulase treatment was reduced by 3.6%, and the CP content was reduced by 12.7%. After 60 days of fermentation, compared with alfalfa raw material, the DM content in the control group (CK) was reduced by 1%, the CP content was reduced by 9.5%, and the WSC (water-soluble carbohydrates) content was reduced by 22.6%. With the cellulase, the lactic acid content of the 30- and 60-day silages was 2.66% DM and 3.48% DM. The content of Firmicutes in salinized alfalfa raw material was less than 0.1% of the total bacterial content. Before and after ensiling, the microbes had similar composition at the phylum level, and were composed of Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria. The abundance of Pantoea was dominant in fresh alfalfa. In the absence of additives, after 30 days and 60 days of silage, the dominant lactic acid bacteria species became Lactococcus and Enterococcus. Conclusions The results showed that LAB (Lactobacillus, Lactococcus, Enterococcus, and Pediococcus) played a major role in the fermentation of saline alfalfa silage. It also can better preserve the nutrients of saline alfalfa silage. The use of cellulase enhances the reproduction of Lactobacillus. The fermentation time would also change the microbial community of silage fermentation.

Flavonoids have attracted the phytochemical scientists attention since their discovery. Today, with the COVID-19 pandemic, have been information about a favorable therapeutic effect in the treatment dynamics of simultaneously taking treatment protocol drugs with traditional Chinese medicine (dietary supplements of plant origin, separate isolated natural compounds) for more than 85% of Chinese patients infected with SARS-CoV-2. Have been presence and content of flavonoids as a class of natural compounds genetically related to the development and accumulation of benzo-α-pyrone derivatives in the aerial part of 20 alfalfa varieties from different breeding sites in the South of Ukraine introduction. The studies were carried out in 2015–2019. Accounting for the main traits and phenological observations of plant development were carried out according to the method field and laboratory research. The cultivation has been carried out on the territory of the southern part left bank Ukraine (on the border of the forest steppe and steppe zones). The raw material for analysis (alfalfa herb) was harvested by cutting off the plants in the flowering stage, dried in the shade, under a canopy, turned over periodically. Solutions for the flavonoids determination were obtained by boiling a crushed raw materials sample with ethanol (70% v/v). Chromatographic separation has been performed on a Shimadzu LC-20 Prominence liquid chromatograph (Shimadzu, Japan) with a Phenomenex Luna C18 column (2); column temperature 35 °C; detection wavelength – from 270 nm to 520 nm; the separation mode is gradient with a constant rate flow of the mobile phase 1 ml/min. The injection volume is 0.5 μl. The compounds presence and content in the alfalfa varieties obtained extracts vary widely enough under the same conditions of sowing, processing, and growing. The Tajik variety Vakhshskaya 233 is the leader in the content of luteolin-7-glucoside. The American variety WL 508 is the richest in routine (0.50920 mg/g). Cinnamic acid was found in maximum quantities in the Mexican variety Tanhuato (0.24406 mg/g), and this component is completely absent in the Ukrainian variety Sinyukha.

Introduction[1] Forages are grown mainly for feeding livestock, especially in dairy cows, because adequate roughage is needed in diets to provide good rumen function. However, as more roughage is fed, the energy density of the diet is reduced. So, the production of high quality forage is very important for dairy producers. High quality forage has direct effects on animal production efficiency, including weight gain, milk production, and reproductive success. Producing and conserving of high quality forage is a challenge because several factors can be affected forage quality including plant species, soil fertility, maturity at harvest, and harvesting (mowing, field curing, baling or chopping) and storage methods and other factors (weeds, insects and diseases). Fiber and energy contents are the most important in forage quality measures. As the fiber level increases, the energy content generally decreases. Therefore, improving forage quality can be achieved by managing forage carbohydrate content. Carbohydrates are the primary source for ruminants and contribute 60 to 70% of the net energy used for milk production and are classified as structural and non-structural. As usual, structural carbohydrates defined as neutral detergent fiber (cellulose, hemicellulose, lignin and portion of the pectin) and non-structural carbohydrates consist of the sugars, starches and pectin. Non-structural carbohydrates are a highly digestible energy source and together with degraded protein, are needed by the rumen for microbial growth and digestion. Plants accumulate sugars during the day via photosynthesis, but incur a net loss at night via dark respiration. This diurnal cycling reflects the concentration of total nonstructural carbohydrates in forages. The aim of this study was to evaluate the effect of morning versus afternoon cutting time on chemical composition, gas production parameters and digestibility of sun-drying alfalfa, clover and barley forages. Materials and Methods About 5-7 cm above the soil stage, alfalfa, clover and barley were harvested in two times, at 06:00 AM and 18:00 PM. Whole Alfalfa and clover plants were harvested at the first bud stage of development and whole barley plant at the medium dough stage of maturity used. Their nutritive value was evaluated through the determination of chemical compositions and in vitro gas production techniques. Samples were tested in an in vitro gas production method (96 h incubation) and batch rumen culture system (24 h incubation). Rumen fluid was collected before the morning feed from three fistulated Dalagh male sheep (45 ± 1.5 kg live weight fed on a forage diet at a concentration of 40:60). In vitro gas production was measured in triplicate and for each replicate; a sample of 200 mg DM was used. The bottles were then filled with 30 ml of incubation medium that consisted of 10 ml of rumen fluid plus 20 ml of buffer solution and placed in a water bath at 39 °C. Gas production was recorded at 2, 4, 8, 16, 24, 48, 72 and 96 h. Total gas values corrected for blank incubation and gas values expressed in ml g-1 of DM. The asymptotic gas production system (A) and rate of gas production (c), organic matter digestibility (OMD), metabolizable energy (ME) and short chain fatty acids (SCFA). A medium similar to one developed for gas production was used for batch rumen culture system to measure pH, and NH3-N and in vitro digestibility. The pH of the media was measured after 24 h incubation. After 24 h incubation, the contents of each glass bottle were empty, strained through four layers of cheesecloth and then 10 ml of strained rumen fluid was acidified by 10 ml of 0.2 N HCl for determination of NH3-N using the distillation method. Finally, all contents remaining in the bottles were filtered through nylon bags, oven dried at 60 °C for 48 h and analyzed for IVDMD and IVOMD. Results and Discussion Results showed that cutting time affected chemical composition of alfalfa and clover forages significantly (p < 0.05), but had not effect on barley forage. Afternoon cutting forages had lower content of NDF and ADF and higher levels of starch and WSC compared to morning cutting forages. There were no significant differences between afternoon and morning cutting forages on gas production parameters (P > 0.05). However, Afternoon cut forages had higher gas production potential than morning cutting forages. Results showed that harvest time had significant effect on DMD (66 vs 59) and OMD (64 vs 58.5) of barley forage. Although, time harvesting had no effect on MCP, EMCP and PF (P>0.05), but Afternoon cut forages had higher MCP, EMCP and PF than morning cutting forages. Conclusion Generally, it was concluded that with considered all factors that affected quality and nutritive value of forages, delaying forage harvest until late afternoon could result in improve nutritive value of forage.

Objective The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p

This study looked at the synergistic effect of Pseudomonas alkylphenolica PF9 and Sinorhizobium meliloti Rm41 on the Moroccan alfalfa population (Oued Lmaleh) grown under symbiotic nitrogen fixation and limited phosphorus (P) availability. The experiment was conducted in a growth chamber and after two weeks of sowing, the young seedlings were inoculated with Sinorhizobium meliloti Rm41 alone or combined with a suspension of Pseudomonas alkylphenolica PF9. Then, the seedlings were submitted to limited available P (insoluble P using Ca3HPO4) versus a soluble P form (KH2PO4) at a final concentration of 250 μmol P·plant-1·week-1. After two months of P stress, the experiment was evaluated through some agro-physiological and biochemical parameters. The results indicated that the inoculation of alfalfa plants with Sinorhizobium strain alone or combined with Pseudomonas strain significantly (p

Chromosome line and characterization of nine Turkish alfalfa (Medicago sativa L.) lines were investigated using karyological techniques in this study. Root tips were obtained from germinated alfalfa seeds in petri dishes at room temperature (25°C) for visualizing somatic chromosomes. The chromosome numbers of nine alfalfa lines were determined as 2n=4x=32. The karyotype formula of 2n=32= 30m+ 2sm (30 median + 2 submedian) were noted for Line 1, 2and 3. Whereas, Line 4, 6, 8, 9 showed the karyotype formula of 2n=32=32m (32 median). The karyotype formula of Line 5 and 7 were 2n=32=28m+4sm (28 median + 4 submedian). Satellite chromosome pairs were observed from Line 5. The results of the karyotype asymmetry index analysis showed that intrachromosomal asymmetry is higher than interchromosomal asymmetry in the karyotypes of alfalfa lines.

Argon, a non-polar molecule, easily diffuses into deeper tissue and interacts with larger proteins, protein cavities, or even receptors. Some of the biological effects of argon, notably its activity as an antioxidant, have been revealed in animals. However, whether and how argon influences plant physiology remains elusive. Here, we provide the first report that argon can enable plants to cope with salinity toxicity. Considering the convenience of the application, argon gas was dissolved into water (argon-rich water (ARW)) to investigate the argon’s functioning in phenotypes of alfalfa seed germination and seedling growth upon salinity stress. The biochemical evidence showed that NaCl-decreased α/β-amylase activities were abolished by the application of ARW. The qPCR experiments confirmed that ARW increased NHX1 (Na+/H+ antiporter) transcript and decreased SKOR (responsible for root-to-shoot translocation of K+) mRNA abundance, the latter of which could be used to explain the lower net K+ efflux and higher K accumulation. Subsequent results using non-invasive micro-test technology showed that the argon-intensified net Na+ efflux and its reduced Na accumulation resulted in a lower Na+/K+ ratio. NaCl-triggered redox imbalance and oxidative stress were impaired by ARW, as confirmed by histochemical and confocal analyses, and increased antioxidant defense was also detected. Combined with the pot experiments in a greenhouse, the above results clearly demonstrated that argon can enable plants to cope with salinity toxicity via reestablishing ion and redox homeostasis. To our knowledge, this is the first report to address the function of argon in plant physiology, and together these findings might open a new window for the study of argon biology in plant kingdoms.

Greater consumer demands for nutritious and healthy foods matching today’s lifestyle requirements are the driving force for the development of innovative ingredients with versatile functionalities. This task becomes even more important when it comes to the part of the population with coeliac disease (CD) whose diet is characterised by high input of fat and carbohydrates but low input of dietary fibres, minerals (Ca, Mg, Fe, Zn) and vitamins. Corresponding nutrient deficiencies can further lead to the development of other related diseases compromising the overall well-being of CD patients. Furthermore, other conditions linked to gluten ingestion and a great number of individuals embracing the gluten-free diet as a healthier choice are other reasons for addressing this global challenge. High nutritional value (rich in proteins, dietary fibres, essential polyunsaturated fatty acids, minerals, vitamins and associated total phenols), absence of gluten, extensive cultivation and availability make lucerne (Medicago sativa L.) or alfalfa seeds an ideal candidate for application in gluten-free bread (GFB) production. However, the presence of antinutrients and beany flavour could limit its use. To tackle these limitations, germination as a natural processing technique is successfully employed enabling also improvements in digestibility of starch and proteins and bioavailability of the other nutrients and bioactive compounds. Considering the presented background, this study aims to explore the effect of non-germinated and germinated alfalfa seeds flour addition (5% on maize flour/starch basis) on GFB mineral composition, protein content and sensory properties (specific volume, external appearance, crumb appearance, smell and taste). Additionally, the aforementioned properties of GFB containing alfalfa were compared with control (without alfalfa) and commercial GFB available on the market. In GFB containing alfalfa significant increase in Mg (from 342.2 to 908.3 mg/kg), K (from 968.8 to 1478.9 mg/ kg) and Zn (from 10.8 to 21.1 mg/kg) content was observed when compared to commercial GFB. However, corresponding bread samples showed lower content of Ca and Fe compared to commercial GFB, but Fe content in GFB containing alfalfa was still higher than in control. Protein content ranged between 4.7 and 6.0 % db and it was higher in GFB containing alfalfa. Control GFB received the highest scores upon sensory evaluation followed by GFB containing germinated and non-germinated alfalfa. Despite the observed volume depressing effect, GFB with alfalfa were characterised by fine taste, soft crumb texture with medium size air pores of good uniformity., This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia [number 451-03-68/2022-14/200134].

Alfalfa is the most important forage legume with symbiotic nitrogen-fixing nodule in roots, but it is sensitive to aluminum (Al), which limits its plantation in acidic soils. One rhizobia clone of Sinorhizobium meliloti with Al tolerance (AT1) was isolated from the nodule in AlCl3-treated alfalfa roots. AT1 showed a higher growth rate than the standard rhizobia strain Sm1021 under Al-stressed conditions. Alfalfa growth was improved by inoculation with AT1 under Al-stressed conditions, with increased length and fresh weight in shoots and roots. High nitrogenase activity and pink effective nodules were obtained in AT1-inoculated plant roots under Al stress, with increased total nitrogen compared with the non-inoculated control. The application of exogenous NH4+-nitrogen increased the Al resistance in alfalfa. It is suggested that rhizobia’s increase of the Al resistance in alfalfa is associated with its improved nitrogen status. Inoculation with Al-tolerant rhizobia is worth testing in an acidic field for improved alfalfa productivity.

Alfalfa (Medicago sativa L.) is a perennial leguminous forage cultivated globally. Fusarium spp.-induced root rot is a chronic and devastating disease affecting alfalfa that occurs in most production fields. Studying the disease resistance regulatory network and investigating the key genes involved in plant–pathogen resistance can provide vital information for breeding alfalfa that are resistant to Fusarium spp. In this study, a resistant and susceptible clonal line of alfalfa was inoculated with Fusarium proliferatum L1 and sampled at 24 h, 48 h, 72 h, and 7 d post-inoculation for RNA-seq analysis. Among the differentially expressed genes (DEGs) detected between the two clonal lines at the four time points after inoculation, approximately 81.8% were detected at 24 h and 7 d after inoculation. Many DEGs in the two inoculated clonal lines participated in PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI) mechanisms. In addition, transcription factor families such as bHLH, SBP, AP2, WRKY, and MYB were detected in response to infection. These results are an important supplement to the few existing studies on the resistance regulatory network of alfalfa against Fusarium root rot and will help to understand the evolution of host–pathogen interactions.

Fodder crops play an important role in sustainable agriculture as they provide feed for animals, which is ultimately converted to human food. Alfalfa is one of the most important fodder crops having high nutritive value for livestock. However, seed production of alfalfa crop is seriously affected by several factors and the highest reduction in seed yield is caused by stink bug infestation. The current study evaluated different insecticides to control stink bugs during 2016–17. The efficacy of ten insecticides, i.e., acephate, dimethoate, malathion, chlorpyriphos, bifenthrin, lambdacyhalothrin, deltamethrin, acetamiprid, imidacloprid and carbosulfan was tested on Agonoscelis spp. (Heteroptera Pentatomidae). The mortality of stink bug was recorded at one, three, five, seven, ten and fifteen days after insecticide application. Similarly, the population of pollinators was recorded before and one, three and five days after the application of insecticides. It was observed that acetamiprid (81.14%) and acephate (80.65%) caused the highest mortality of stink bug and proved most effective. Insecticides application decreased the pollinators’ population one day after spray; however, it was rehabilitated three days after insecticide application. Insecticide application increased seed yield from 28.05 kg/acre (during last four year without chemical control) to 116 kg/acre in 2016–17 (with chemical control). It is concluded that acetamiprid and acephate can be successfully used in integrated management program of increasing alfalfa seed production.

Alfalfa (Medicago sativa L.) is an important forage legume grown in Kansas, USA and its productivity with cool-season grasses however is unknown. The objective of this study was to determine the dry matter yield (DMY) and forage nutritive value of alfalfa-grass mixtures compared to those of alfalfa and grasses grown in monoculture with and without nitrogen fertilization. Three different alfalfa varieties were planted (reduced-lignin alfalfa, Roundup Ready, and conventional alfalfa) and two kinds of cool-season grasses (smooth brome, Bromus inermis Leyss, and tall fescue, Festuca arundinacea Schreb) were planted as a monoculture or in alfalfa-grass mixtures. Nitrogen fertilizer (urea) was applied at green-up at a rate of 56 kg/ha and after the second cutting at a rate of 56 kg/ha in 2016 and 2017, respectively. and control treatments received no nitrogen. DMY was significantly higher in monoculture alfalfa and alfalfa-grass mixtures than in grass monocultures. Between alfalfa monoculture and alfalfa-grass mixtures, no significant differences in DMY were found. For all treatments, nitrogen application significantly increased DMY compared to the control. In 2016 and 2017, the low-lignin alfalfa monoculture had the lowest acid detergent fiber (ADF) and the grass monocultures had the highest ADF. In 2016 and 2017, neutral detergent fiber (NDF) in smooth bromegrass and tall fescue was higher than in other species treatments. A low-lignin alfalfa monoculture had significantly lower NDF concentration compared to alfalfa-grass mixtures. When averaged over 2016 and 2017, relative feed value (RFV) was highest in low-lignin alfalfa and lowest in the grass monocultures. In both years, nitrogen fertilizer application did not affect nutritive values.

Background Alfalfa (Medicago sativa L.) is a perennial legume extensively planted throughout the world as a high nutritive value livestock forage. Flowering time is an important agronomic trait that contributes to the production of alfalfa hay and seeds. However, the underlying molecular mechanisms of flowering time regulation in alfalfa are not well understood. Results In this study, an early-flowering alfalfa genotype 80 and a late-flowering alfalfa genotype 195 were characterized for the flowering phenotype. Our analysis revealed that the lower jasmonate (JA) content in new leaves and the downregulation of JA biosynthetic genes (i.e. lipoxygenase, the 12-oxophytodienoate reductase-like protein, and salicylic acid carboxyl methyltransferase) may play essential roles in the early-flowering phenotype of genotype 80. Further research indicated that genes encode pathogenesis-related proteins [e.g. leucine rich repeat (LRR) family proteins, receptor-like proteins, and toll-interleukin-like receptor (TIR)-nucleotide-binding site (NBS)-LRR class proteins] and members of the signaling receptor kinase family [LRR proteins, kinases domain of unknown function 26 (DUF26) and wheat leucine-rich repeat receptor-like kinase10 (LRK10)-like kinases] are related to early flowering in alfalfa. Additionally, those involved in secondary metabolism (2-oxoglutarate/Fe (II)-dependent dioxygenases and UDP-glycosyltransferase) and the proteasome degradation pathway [really interesting new gene (RING)/U-box superfamily proteins and F-box family proteins] are also related to early flowering in alfalfa. Conclusions Integrated phenotypical, physiological, and transcriptomic analyses demonstrate that hormone biosynthesis and signaling pathways, pathogenesis-related genes, signaling receptor kinase family genes, secondary metabolism genes, and proteasome degradation pathway genes are responsible for the early flowering phenotype in alfalfa. This will provide new insights into future studies of flowering time in alfalfa and inform genetic improvement strategies for optimizing this important trait.

Background: Lutein and its isomerzeaxanthin are safe natural compounds. They are able to reduce the development of tumor and other chronic diseases. The objectives of the current study was to examine the cytotoxicity of lutein isolated and purified from alfalfa, safe and low-cost plant, on five different human cancer cell lines, namely (MCF-7), (HepG2), (A549), (PC3), and (HCT116), as well as normal (HFB4) cells. Method: We examined the cytotoxicity of lutein purified from Medicago sativa L and evaluate its activity against human liver HepG2, breast MCF-7, lung A549, prostate PC3, and colon HCT116 cancer cell lines using SRB assay in comparison with doxorubicin as a reference drug. Results: Results revealed that the tested extract could be a more promising anticancer agent in the case of MCF-7 (IC50, 3.10±0.47 μg/ml) compared with standard drug doxorubicin (IC50, 2.90±0.30 μg/ml). Moreover, the extract showed a moderate effect on HepG2 (IC50, 6.11±0.84 μg/ml) versus doxorubicin (IC50, 2.90±0.30 μg/ml); meanwhile, the extract showed no activity against A549, PC3, and HCT116 cells. The results further revealed that the extract had no toxicity against the growth of normal HFB4 cells versus doxorubicin. Conclusion: Lutein-rich extract from alfalfa had a major antiproliferative role in breast MCF-7 and liver HepG2 compared to doxorubicin.

Ruminants would suffer from gastric distension (bloat) when feeding on alfalfa (Medicago sativa L.; Alf), while tannins can relieve this problem. Thus, this study aimed to evaluate the effect of Alf mixed with paper mulberry (Broussonetia papyrifera) leaves (PML) or condensed tannins (CT) extracted from PML on in vitro fermentation performance. Seven experimental treatments based on the mass ratios of Alf and PML were set up: Alf, PML, 3A1P (Alf:PML = 3:1), 1A1P (Alf:PML = 1:1), 1A3P (Alf:PML = 1:3), A1P (Alf with 1% CT extracted from PML), A2P (Alf with 2% CT extracted from PML), and a blank. The results after fermentation were as follows. (1) The degradable dry matter (DDM) and degradable organic matter (DOM) contents increased simultaneously with the increase in the proportion of PML (p .05), propionic, and butyric acids compared to Alf alone. Adding CT improved the concentrations of propionic and butyric acids over the Alf treatment (p

The objective of this study was to evaluate the effects of lactic acid bacteria (LAB) inoculants on fermentation quality and subsequent in vitro gas production, ruminal fermentation parameters, cellulolytic bacteria and their activities of alfalfa silage. Primary growth of alfalfa (Medicago sativa L.) was harvested at 50% flowering stage, inoculated without (control) or with Lactobacillus plantarum, Enterococcus mundtii and Enterococcus faecalis at 1.0 × 106 cfu/g of fresh weight (FW) in quadruplicate laboratory silos for 45 d. The silage inoculated with LAB were well preserved, indicated by the lower (p

Aim. Alfalfa is a rich source of phytoestrogens, among them coumestrol which shows strong estrogenic activity that can adversely affect the health of domestic animals. The aim of the study was to determine the variation in coumestrol content in leaves of alfalfa breeding populations, present in the breeding program of Agricultural Institute Osijek in Croatia. Method. Twelve alfalfa populations were selected based on their high forage yield and good persistence. Coumestrol was extracted using acidifi ed methanol as an organic solvent from lyophilized and ground alfalfa leaves, while for detection and quantifi cation was used. Results. Signifi cant differences were observed between the studied populations with average coumestrol content of 435.67 mg/kg of dry matter (DM). The highest content of coumestrol was determined in breeding population Rs-21 (619.53 mg/kg of DM). Conclusions. Populations Rs-33 and Rs-20 had the lowest coumestrol content (82.18 and 86.58 mg/kg, respectively) and present a potential breeding source for creat- ing new contemporary cultivars with decreased coumestrol content

Background and Objective: The goal of study was to evaluate promising alfalfa cultivars for drought stress and identify the most tolerant and superior cultivars for drought stress. Materials and Methods: The experiment was conducted as split plot based on randomized complete blocks design with three replications in the East Azarbaijan Agriculture and Natural Resources Research and Education Center during three cropping seasons (2016-19). In the main plots, the irrigation factor was applied at three levels - at the first irrigation level only to the end of the first harvest, at the second level irrigation until the end of the second harvest, and at the third level irrigation until the end of the third harvest (complete irrigation). The second factor was six cultivars and alfalfa ecotypes. Results: The effect of irrigation levels was significant for all measured traits. The fresh and dry matter yields of the studied genotypes were affected by irrigation levels and decreased significantly. Water stress significantly reduced plant height at first and second levels of irrigation 23% and 17%, respectively. Water stress, also significantly reduced the number of stems at the first and second levels of irrigation 34% and 30%, respectively. Conclusion: The responses of genotypes for dry matter yield were similar as fresh forage yield. In conclusion, synthetic and Azar cultivars and Varzghan ecotype were the most suitable cultivars for deficit irrigation conditions and are suitable for cultivation in low irrigation conditions in Tabriz plain.

Background: This research aims to evaluate heavy metals contamination of Alfalfa forage and soil in industrial and non-industrialregions of Tehran province. The soil and plant samples were collected from Alfalfa fields situated in industrial and non-industrialareas during two harvests of Alfalfa harvest.Methods: The Electrothermal atomizers spectroscopy technique was employed to measure the heavy metals (lead, arsenic, andcadmium) levels in the samples. A flame atomic absorption spectrometer equipped with acetylene-air flame was employed.Results: The highest heavy metals concentrations were detected in the soil and Alfalfa samples from the industrial regions. Lead hadthe highest level in the soil followed by arsenic and cadmium; the same trend held for the Alfalfa as well. Moreover, the heavy metalcontent in Alfalfa declined from the first to second harvest, while it increased in the soil. The heavy metals content of Alfalfa wasbelow the maximum permitted concentration for feedstuffs and livestock. The highest soil-plant transfer factor of heavy metals wasdetected in the first harvesting. The highest arsenic transfer factor was recorded in the industrial areas.Conclusion: The findings of the current study can be helpful in the determination of healthy forage in various areas

Copper metal as a contact surface was studied during the germination of alfalfa seeds (Medicago sativa L.) inside a rotating drum on a laboratory scale and compared with a plastic surface of food-grade. A system of three rotating drums was used inside a thermo-regulated chamber to germinate seeds. Alfalfa seeds were inoculated with 4.2 log cfu g-1 of Escherichia coli and after 84 h of germination sprouts were evaluated for E. coli, mesophilic aerobic bacteria, the content of copper and other minerals (potassium, calcium, magnesium, sodium, iron, manganese, and zinc), total mass, unit mass and length, and color. The contact of alfalfa sprouts with copper sheets allowed to reduce the E. coli load from 6.54 to 10 ppm) according to Food Sanitary Regulations. Germinated mass and length decreased after copper treatments. No statistically significant differences were observed between treatments for the remaining quality parameters. Finally, it is concluded that copper was very efficient in reducing the microbial load of E. coli in alfalfa sprouts, complying with the regulations established by the Chilean Ministry of Health.

The results of field studies of nitrogen consumption in alfalfa crops depending on the nitrogen nutrition pattern and symbiotic activity of crops for 2017...2019 are presented. Experiments with the alfalfa purple were carried out in the ecological conditions of the foothill zone of the North Ossetia-Alania, on leached chernozem with a close occurrence of gravel. Mineral and symbiotrophic patterns of nitrogen nutrition of alfalfa plants were compared. It was found that in the year of planting, the nitrogen consumption of alfalfa crops ranged from 105.0 to 135.4 kg/ha, and by the first mowing, the consumption was 1.1...1.2 times bigger than by the second mowing. In the second and third years of crop use, nitrogen consumption increased by 1.7...1.9 times compared to the year of planting. The parameters of the options with natural conditions (control) and the addition of starting doses of nitrogen fertilizers (N30) were almost identical. In the second and third years of use of crops, parameters of options with inoculation of seeds with high-altitude inoculum (In-1800) and the addition of starting doses of nitrogen fertilizers within the background of inoculation (N30 + in) were almost identical. In these options, the amount of nitrogen consumption was more than 235 kg/ha, or 25.6...27.1 % higher than in the control variant. In total, over three years of research, in the control option alfalfa crops took 483.3 kg/ha of nitrogen. Starting doses of nitrogen fertilizers (N30) increased this indicator by only 1.4 %. Pre-planting inoculation of seeds contributed to an increase in nitrogen consumption in all options: in the option with the industrial strain of risotorphine (St. 425a) – by 17.8 %, with high-altitude inoculum within the background of adding starting doses of mineral nitrogen fertilizers (N30 + In) – by 24.6 %, with high-altitude inoculum in pure form (In-1800) – by 27.1 %., Volga Region Farmland, Выпуск 3 (7) 2020, Pages 37-41

Alfalfa (Medicago sativa L.) is the most widely grown forage legume crop worldwide. Yield and plant height are important agronomic traits influenced by genetic and environmental factors. The objective of this study was to identify quantitative trait loci (QTL) and molecular markers associated with alfalfa yield and plant height. To understand the genetic basis of these traits, a full-sib F1 population composed of 392 individuals was developed by crossing a low-yielding precocious alfalfa genotype (male parent) with a high-yielding late-maturing alfalfa cultivar (female parent). The linkage maps were constructed with 3818 single-nucleotide polymorphism (SNP) markers on 64 linkage groups. QTL for yield and plant height were mapped using phenotypic data for three years. Sixteen QTL associated with yield and plant height were identified on chromosomes 1 to 8. Six QTL explained more than 10% of phenotypic variation, representing major loci controlling yield and plant height. One locus on chromosome 1 controlling yield traits had not been identified in previous studies. Three QTL co-located with other QTL (qyield-1 and qheight-7, qheight-5 and qyield-4, qheight-6, and qyield-6). With further validation, the markers closely linked with these QTL may be used for marker-assisted selection in breeding new alfalfa varieties with high yield.

Arising incidence of coeliac disease made both consumers’ and food industry aware of the close link between diet and health far beyond the well-known role of food in satisfying hunger. The emerging requirements for healthy gluten-free foods providing necessary nutrients has steadily prompted the research and development towards exploration and introduction of novel gluten-free raw materials as an alternative to refined flours and starches. However, finding the right balance between nutritive value and technological functionality of such gluten-free raw materials creates many challenges nowadays especially in production of gluten-free bread that will match the properties of its wheat counterparts. As one of the extensively cultivated forage legume alfalfa or lucerne (Medicago sativa L.), and its seeds possessing high nutritional value (rich in proteins, dietary fibres, essential polyunsaturated fatty acids, vitamins and associated total phenols) have immense potential as a gluten-free food resource. In application of novel gluten-free flours as baking ingredients, knowing the water absorption properties of flour represents one of the essential prerequisites for reaching the final product with desired overall quality. Additionally, differences in novel gluten-free flours effect on the final product quality can be predicted to some extent by assessing its hydration properties. In this regard, presented study aims to investigate and compare the hydration (water binding capacity, water holding capacity, and swelling) and oil binding properties of grounded raw alfalfa seeds, grounded germinated alfalfa seeds, and alfalfa sprouts powder. The highest water holding capacity and swelling was recorded for alfalfa sprouts 6.48 g/g and 7.13 cm3/g, respectively, whilst germinated alfalfa seeds had the highest water binding capacity (3.13 g/g). Oil binding capacity ranged from 0.90 to 1.41 g/g with the highest value reported for alfalfa sprouts followed by germinated and raw alfalfa seeds. The obtained results demonstrated that alfalfa sprouts, considering their hydration properties, can have a great impact on other gluten-free ingredients functionality, product yield and shelf stability, while in terms of oil binding can act as flavour retainer increasing the mouth feel of gluten-free bakery products. Additionally, germinated and raw alfalfa seeds also showed hydration and oil binding properties desirable for the gluten-free bread production. Further research should be conducted to establish the raw and germinated alfalfa seeds and sprouts influence on gluten-free bread technological, sensory and nutritional quality.

Within Mediterranean cropping systems, legume forage crops that last up to 6 years or more (e.g., alfalfa) are replaced with cereal crops (e.g., wheat). The change from forage to cereal crops has negative climate and environmental impacts that must be addressed with mitigation actions. This study evaluated the synergies and tradeoffs between the ecosystem services provided by three management systems after forage legume. A field trial was set up from October 2017 to September 2019 on a 6-year-old alfalfa field subjected to the following management systems: (i) alfalfa termination followed by wheat for 2 years (WW, control); (ii) alfalfa termination followed by single amendment with 60 Mg ha−1 recalcitrant biochar and then by wheat for 2 years (WWB60); and (iii) extension of alfalfa for 2 years (AEXT). A range of regulating, supporting, and provisioning ecosystem services were assessed during the 2018 and 2019 cropping seasons. The results highlight that WWB60 can guarantee carbon sequestration without causing tradeoffs with other services, while AEXT can enhance soil conservation while not increasing soil greenhouse gas emissions. Future policies should support the WWB60 system if the goal is to increase the supporting services. Conversely, the AEXT system should be used if the goal is to increase the regulating and provisioning services.

Interseeding silage corn into an alfalfa stand following its spring harvests to produce mixed silage would feasibly break the restriction of alfalfa production during a hot-rainy summer. In this 2-yr experiment, the alfalfa–silage corn intercropping system (AC) with nitrogen (N) fertilization rates of 0, 60, 120, and 180 kg N ha−1 and corn plant densities of 30,000, 45,000, and 60,000 plants ha−1 was compared with the monocultured corn (CK) managed using the local farmers’ practice. Intercropping with alfalfa significantly decreased the dry matter (DM) yield of the corn by 31.5% on average, while increasing the N application rate and corn plant density promoted DM yield and crude protein (CP) accumulation of the intercropped corn, and the intercropping system could achieve the same CP yield as CK—it did in most cases. Although the DM contents, pH values, and NH3-N concentrations were decreased in the mixed silage, its fermentation quality was not reduced and the CP, water soluble carbohydrates, and lactic acid contents were dramatically increased by 16.3%, 17.8%, and 72.9% compared with those of CK, respectively. For the opposite effects of N fertilization and corn density on silage quality, the treatment combination with a medium corn density (45,000 plants ha−1) fertilized at a medium N rate (120 kg N ha−1) was evaluated as the optimum for more balanced system productivity, silage nutritive values and fermentative characteristics during the coexisting period.

Medicago sativa(also known as alfalfa), a forage legume, is widely cultivated due to its high yield and high-value hay crop production. Infectious diseases are a major threat to the crops, owing to huge economic losses to the agriculture industry, worldwide. The protein-protein interactions (PPIs) between the pathogens and their hosts play a critical role in understanding the molecular basis of pathogenesis.Pseudomonas syringaepv.syringaeALF3 suppresses the plant’s innate immune response by secreting type III effector proteins into the host cell, causing bacterial stem blight in alfalfa. The alfalfa-P. syringaesystem has little information available for PPIs. Thus, to understand the infection mechanism, we elucidated the genome-scale host-pathogen interactions (HPIs) between alfalfa andP. syringaeusing two computational approaches: interolog-based and domain-based method. A total of ∼14 M putative PPIs were predicted between 50,629 alfalfa proteins and 2,932P. syringaeproteins by combining these approaches. Additionally, ∼0.7 M consensus PPIs were also predicted. The functional analysis revealed thatP. syringaeproteins are highly involved in nucleotide binding activity (GO:0000166), intracellular organelle (GO:0043229), and translation (GO:0006412) while alfalfa proteins are involved in cellular response to chemical stimulus (GO:0070887), oxidoreductase activity (GO:0016614), and Golgi apparatus (GO:0005794). According to subcellular localization predictions, most of the pathogen proteins targeted host proteins within the cytoplasm and nucleus. In addition, we discovered a slew of new virulence effectors in the predicted HPIs. The current research describes an integrated approach for deciphering genome-scale host-pathogen PPIs between alfalfa andP. syringae, allowing the researchers to better understand the pathogen’s infection mechanism and develop pathogen-resistant lines.

This study evaluated the effects of hydrolysable tannin (HT) and condensed tannin (CT) on the bacterial community, fermentation quality, and proteolysis of alfalfa silage. Alfalfa was wilted to a dry matter (DM) of 35% fresh weight and ensiled with or without 4% HT or 4% CT. The application rates of tannins were based on fresh weight, and each treatment was ensiled in triplicate. After 60 d of fermentation, the CT-treated group had lower concentrations of ammonia nitrogen (NH3-N) and free amino acid nitrogen (AA-N), but greater lactic acid concentration, than those in the control and HT-treated silage (p < 0.05). Compared to the control group, the application of tannins increased the abundance of Pseudomonas (negatively correlated with aminopeptidases activity), and decreased the abundance of Pediococcus—which was positively correlated with aminopeptidases activity—and the concentrations of non-protein nitrogen (NPN), NH3-N, and AA-N. The application of HT decreased the abundance of Lactobacillus and increased the abundances of Enterococcus, while the opposite results were observed in the CT-treated group. The application of HT and CT reduced the proteolysis in treated silages, but the two were different in terms of their mechanism and their effects on the bacterial communities of the alfalfa silage.

The study focused on the isolation, characterization and conservation of fungal strains isolated form Medicago sativa plants showing wilt disease symptoms in southern Tunisian oases. A total of 79 fungal strains were isolated from different plant organs, including roots, collars, and stems. Morphological identification of fungal strains was based on the morphology of culture on PDA medium and of the conidia under microscope. All the fungal strains were stored on PDA slant covered with mineral oil at room temperature. Over all, three main fungal genera were identified i.e. Fusarium sp., Rhizoctonia sp., and Alternaria sp. with unidentified strains. These findings demonstrate that alfalfa wilt disease is probably caused by a complex of pathogens rather than the effect of a single pathogen.The study focused on the isolation, characterization and conservation of fungal strains isolated form Medicago sativa plants showing wilt disease symptoms in southern Tunisian oases. A total of 79 fungal strains were isolated from different plant organs, including roots, collars, and stems. Morphological identification of fungal strains was based on the morphology of culture on PDA medium and of the conidia under microscope. All the fungal strains were stored on PDA slant covered with mineral oil at room temperature. Over all, three main fungal genera were identified i.e. Fusarium sp., Rhizoctonia sp., and Alternaria sp. with unidentified strains. These findings demonstrate that alfalfa wilt disease is probably caused by a complex of pathogens rather than the effect of a single pathogen.

The study focused on the isolation, characterization and conservation of fungal strains isolated form Medicago sativa plants showing wilt disease symptoms in southern Tunisian oases. A total of 79 fungal strains were isolated from different plant organs, including roots, collars, and stems. Morphological identification of fungal strains was based on the morphology of culture on PDA medium and of the conidia under microscope. All the fungal strains were stored on PDA slant covered with mineral oil at room temperature. Over all, three main fungal genera were identified i.e. Fusarium sp., Rhizoctonia sp., and Alternaria sp. with unidentified strains. These findings demonstrate that alfalfa wilt disease is probably caused by a complex of pathogens rather than the effect of a single pathogen.The study focused on the isolation, characterization and conservation of fungal strains isolated form Medicago sativa plants showing wilt disease symptoms in southern Tunisian oases. A total of 79 fungal strains were isolated from different plant organs, including roots, collars, and stems. Morphological identification of fungal strains was based on the morphology of culture on PDA medium and of the conidia under microscope. All the fungal strains were stored on PDA slant covered with mineral oil at room temperature. Over all, three main fungal genera were identified i.e. Fusarium sp., Rhizoctonia sp., and Alternaria sp. with unidentified strains. These findings demonstrate that alfalfa wilt disease is probably caused by a complex of pathogens rather than the effect of a single pathogen.

Phosphorus (P) is one of the essential nutrients for alfalfa (Medicago sativa L.) growth, but P deficiency in soil is a common phenomenon. Applying an appropriate amount of P fertilizer for alfalfa in the establishment year improves winter survival, helps achieve sustainable high yields, and promotes the best economic returns. The objective of this study was to determine the effect of different P fertilization rates (0, 22, 44, and 66 kg P ha−1) on winter survival, forage yield, and root traits of two late-summer seeded alfalfa cultivars (dormant and semi-dormant) in the establishment and first production year. Our results showed that applying an appropriate amount of P fertilizer can increase forage yield and enhance the persistence of late-summer seeded alfalfa by promoting the development of crown and root traits and improving winter survival, especially for semi-dormant cultivars. Late-summer seeding and application of appropriate P fertilizer rates are effective agronomic methods for alfalfa in cold and arid winter regions, and can be used as strategies to improve fertility and stabilize soil from wind erosion for sustainable agriculture. To obtain high rate of survival in winter, the optimal P fertilization rates for the dormant and semi-dormant alfalfa cultivars in this study were 31.4 and 41.3 kg P ha−1, whereas the optimum P application rates for high forage yields were 31.3 and 31.5 kg P ha−1, respectively.

The cultivation of alfalfa (Medicago sativa L.), a forage crop grown worldwide, is negatively affected by hard seed presence. We show that treatment of alfalfa seeds with an inductively coupled radio frequency oxygen plasma improves their surface hydrophilicity, as determined by water contact angle (WCA) measurements and water uptake. Furthermore, we see that these effects are mediated by functionalization and etching of the alfalfa seed surface. Surface chemistry is analyzed by X-ray photoelectron spectroscopy (XPS), while morphology is viewed using scanning electron microscopy (SEM). Plasma produces effective alfalfa seed hydrophilization with a variety of treatment parameters. With its potential for fine-tuning, plasma modification of seed wettability shows promise for introduction into agricultural practice.

Alfalfa is widely grown worldwide as a perennial high-quality legume forage and as a good ecological landcover. The cysteine synthase (CSase) gene family is actively involved in plant growth and development and abiotic stress resistance but has not been systematically investigated in alfalfa. We identified 39 MsCSase genes on 4 chromosomes of the alfalfa genome. Phylogenetic analysis demonstrated that these genes were clustered into six subfamilies, and members of the same subfamily had similar physicochemical properties and sequence structures. Overexpression of the CSase gene in alfalfa increased alkali tolerance. Compared with control plants, the overexpression lines presented higher proline, soluble sugars, and cysteine and reduced glutathione contents and superoxide dismutase and peroxidase activities as well as lower hydrogen peroxide and superoxide anion contents after alkali stress. The relative expression of γ-glutamyl cysteine synthetase gene (a downstream gene of CSase) in the overexpression lines was much higher than that in the control line. The CSase gene enhanced alkalinity tolerance by regulating osmoregulatory substances and improving antioxidant capacity. These results provide a reference for studying the CSase gene family in alfalfa and expanding the alkali tolerance gene resources of forage plants.

The alfalfa weevil Hypera postica (Gyllenhal) (Coleoptera: Curculionidae) is a major pest of alfalfa worldwide. In Spain, insecticide sprayings are mainly used for control with low efficacy. As a potential more sustainable control method, we determined the effectiveness of an alfalfa winter cutting for reducing pest populations in the spring and its interaction with parasitism rate due to Bathyplectes spp. (Hymenoptera: Ichneumonidae). Forty-two commercial fields were split in two parts and one was mowed during winter. Larval abundance in each part of the field was sampled by sweep netting in 2019 and 2020 before the first alfalfa spring cutting, when damage is caused. The rates of parasitism due to Bathyplectes anura (Thomson) and Bathyplectes curculionis (Thomson) were estimated by rearing larvae in the laboratory. Winter cutting significantly reduced the spring larval weevil populations and favored the rate of larval parasitism. Our results suggest that winter cutting can be a useful cultural method for alfalfa weevil control that has potential to be a component of an integrated pest management program. This study was funded by Ministerio de Ciencia, Innovación y Universidades, Spanish Government. Project AGL2017-84127-R

Alfalfa (Medicago sativa L.) is one of the most widely cultivated forage crops in the world. China is the second largest producer of alfalfa in terms of the planting area worldwide, with Gansu, Henan, Inner Mongolia, and Shaanxi provinces being the production hubs. Alfalfa viruses have been reported on a small-scale survey in some of these areas, but they have not been well characterized. In the present study, seven viruses were detected in 12 fields of 10 cities/counties of the four abovementioned provinces by high-throughput sequencing and assembly of small RNA. Their incidence, distribution, and genetic diversity were analyzed by enzyme-linked immunosorbent assay, polymerase chain reaction (PCR)/reverse transcription-PCR and clone sequencing. The results showed that alfalfa mosaic virus (AMV), pea streak virus (PeSV), lucerne transient streak virus (LTSV), alfalfa dwarf virus (ADV), Medicago sativa alphapartitivirus 1 (MsAPV1), MsAPV2, and alfalfa leaf curl virus (ALCV) were the main viruses infecting alfalfa in four examined provinces. AMV and MsAPV1 had the highest incidences in all 4 provinces. SDT analysis of the 7 viruses isolated in China revealed a highly conserved among AMV, LTSV, ADV, MsAPV1, MsAPV2, and ALCV, but the sequence was a high variation between China isolates to abroad isolates in PeSV, ADV, and ALCV. To our knowledge, this is the first report of ADV in Inner Mongolia and Gansu, ALCV in Inner Mongolia, MsAPV1 and MsAPV2 in all 4 provinces, and PeSV and LTSV in China. These findings provide a basis for future research on the genetic evolution of alfalfa viruses in China and on strategies to prevent diseases in alfalfa caused by these viruses.

As the population increases and available land for food production decreases, it is necessary for livestock producers to continually work towards increasing livestock production efficiency. In livestock operations, feed accounts for the majority of input costs associated with raising livestock. As such, it is necessary to improve growth and production of livestock animals, while also optimizing feed utilization. Different feedstuffs can be included in the diet of livestock animals to maximize growth and production. However, the effects of some of these novel feedstuffs on growth and production of livestock animals has not been elucidated. As such, we investigated the effects of including two novel alfalfa products, ProLEAF MAX��� (a pellet composed of alfalfa leaves) and ProFiber Plus��� (alfalfa stems), in the diets of beef steers, dairy heifers, and lactating dairy cows. We hypothesized that inclusion of ProLEAF MAX��� and ProFiber Plus��� in the diet would result in improved growth and performance of beef steers, growth and development of dairy heifers, and milk yield and milk components of lactating dairy cows. We found that inclusion of ProFiber Plus��� in the diet of beef steers and dairy heifers decreases feed costs without affecting overall growth in steers, but decreases growth in dairy heifers and inclusion of the two novel alfalfa products in the diet of lactating dairy cows results in improved milk yield and milk components. Additionally, we examined the effects of supplementing murine myoblasts with polyamines and polyamine precursors to further investigate novel products that may be able to be utilized in the diets of livestock animals to increase growth. We hypothesized that supplementation of polyamines and their precursors would result in improved growth of skeletal muscle cells (myoblasts). Treatment of myoblasts with polyamines and their precursors improves proliferation rates and alters mRNA expression of genes involved in polyamine biosynthesis, cell proliferation, and protein synthesis. Collectively, our observations suggest that various novel feedstuffs, whether it be alfalfa processed differently or amino acid derivatives (polyamines), have the potential to improve various growth and/or production measures. However, additional research is required to fully understand the potential of including these products in the diet.