Alfalfa Seeds Research Articles

Seed Yield, Seed Attributes and Economics of Alfalfa Seed Production as Influenced by Integrated Nutrient Management During kharif Season under Southern Dry Zone of Karnataka, India

The field experiment was carried out at the farm field of Krishi Vigyan Kendra, Konehalli, Tiptur taluk, Tumkuru, Karnataka during Kharif season of 2017-18 to examine the effect of integrated nutrient management on seed yield, seed attributes and net benefit of alfalfa. The experiment was laid out Randomized complete block design (RCBD) comprising nine treatments viz T1 : RDF (25:50:25 kg NPK/ha + 10 t/ha FYM); T2 : 75% RDF + 25% N through FYM; T3 :75% RDF + 25% N through Vermicompost; T4 :75% RDF + 25% N through Poultry manure; T5 : 50% RDF + 25% N through FYM + Rhizobium + PSB+VAM ; T6 :50% RDF + 25% N through Vermicompost+ Rhizobium + PSB+VAM; T7 :50% RDF + 25% N through Poultry manure+ Rhizobium + PSB+VAM ;T8 : RDF + Rhizobium + PSB+VAM ;T9 : 10 t/ha FYM + 100% N through FYM and replicated four times. The results of experiments revealed that the plants supplied with of 50 % RDF + 25 % N through vermicompost + Rhizobium + PSB + VAM shown highest fresh herb yield at first harvest (160.65 q/ha), herb yield after harvest of seeds (187.50q/ha), 50% flowering (35.00 days), pod initiation (48.50days), pod maturation (73.05 days), while, lower fresh herb yield, pod initiation, pod maturation were recorded when 10 t/ha FYM + 100 % N through FYM was applied. The maximum pod (56.78 %), number of pods (228.75/plant), pod yield (19.90/plant), pod length (6.25 mm), pod weight (87.61 mg), seed set (56.50 %), number of seeds (5.50/pod), test weight (3.33g), seed yield per hectare (223.38kg/ha) and highest net return per hectare were recorded with 50 % RDF + 25 % N through vermicompost + Rhizobium + PSB + VAM. while, poor performance with regard to these parameter were recorded with application of 10 t/ha FYM + 100 % N through FYM. Therefore, application of 50 % RDF + 25 % N through vermicompost + Rhizobium + PSB + VAMmay be recommended for commercial seed productionin Alfalfa during kharif season under southern dry zone of Karnataka.

Impact of Ozone Exposure on the Biochemical Composition of Wheat, Broccoli, Alfalfa, and Radish Seeds During Germination

In recent years, there has been an increasing interest in the use of gaseous ozone (O3) to promote the germination of edible seeds. While its ability to improve seedling vigor and stimulate germination is acknowledged, there has been limited research on the impact of gaseous O3 on the biochemical profile, including phenolic compounds (TPC) and antioxidant activity, of cereals, seeds, and their sprouts. The lack of information has led to the initiation of this study, which aims to assess the impact of ozone treatment duration at a concentration of 50 ppm 1 L min−1, ranging from 1 to 5 h, on the biochemical attributes of broccoli (Brassica oleracea), radish (Raphanus sativus), alfalfa (Medicago sativa) seeds, and sprouts, as well as wheat (Triticum aestivum) grains and sprouts. By optimizing O3 exposure parameters, including duration, this approach has the potential to serve as a valuable tool for enhancing the microbiological and nutritional quality of seeds and cereals. The findings revealed that O3 treatment generally had an adverse impact on TPC in seeds, cereals, and sprouts, resulting in a significant reduction in TPC post O3 treatment. Wheat grains, in particular, displayed the lowest TPC following ozone exposure, with an average decrease of 39.4% compared to the untreated sample. However, it is noteworthy that alfalfa seeds exhibited a positive response to 4 and 5 h O3 treatment, manifesting an average increase in TPC of 13.0% and 27.7%, respectively. In turn, broccoli, radish, and wheat sprouts displayed the lowest TPC, with values of 47.7%, 20.2%, and 18.0% lower than the control samples, respectively. This study revealed that plant responses to O3 exposure varied, and the effects of O3 treatment on TPC levels depended on O3 exposure time. Furthermore, the effect of O3 on the sugar content of the seeds, cereals, and sprouts varied among different plant types, with some showing an increase in content and others showing no substantial changes. This suggests that, depending on the type of seed, O3 may have both positive and neutral effects.

РАЗРАБОТКА ТЕХНОЛОГИЧЕСКОЙ СХЕМЫ И ОБОСНОВАНИЕ ПАРАМЕТРОВ ОЧИСТКИ СЕМЯН ЛЮЦЕРНЫ

Despite the high effectiveness of chemical preparations against weeds, this fight begins with the preparation of seed material on seed cleaning lines. An urgent problem in cleaning crop seeds is the isolation of dodder seeds in the preparation of alfalfa seeds. Alfalfa cleaning is a time-consuming process in itself, due to the complex structure of the combine heap. In this regard, it is important to develop a technological scheme for cleaning the alfalfa heap, which will allow you to effectively separate dodder seeds on a par with other impurities. When developing the technological scheme for alfalfa purification, the parameters of the initial heap and its components were determined. When determining the composition of the alfalfa heap, it was found that the alfalfa heap consists of 40-60% of organic impurities, 10% of puny seeds, and up to 10% of the heap consists of unthreshed beans, and the alfalfa seeds themselves in the initial heap are usually no more than 30% of the total weight, it was also determined that weed seeds make up less than 1% of the original weight, nevertheless, the heap contained 0.02% of dodder seeds. On laboratory screens with slotted notches and round holes, the dimensional characteristics of the components are determined. On the sail classifier, the distributions of the components according to the critical speed of vinow were established. The data obtained made it possible to develop a technological scheme for cleaning alfalfa seeds, which allows you to effectively remove dodder seeds with minimal losses. Using the same equipment in laboratory conditions, the proposed alfalfa cleaning scheme was tested, which showed that by combining aspiration and sieve cleaning, it is possible to achieve 99.45% purity of seeds, with losses of no more than 1%, dodder was not found in the final material.

Effect of Treating Alfalfa Seeds with Gibberellin and Nano-fertilizer on Productive Characteristics and Protein Content in Hydroponic Cultures

This study was carried out in an experiment in an incubator designed for hydroponic seed germination with tight thermal insulation in the laboratories of the Department of Plant Production Technologies at the Technical College of Agriculture in Mosul, which is affiliated with the Northern Technical University in Iraq, where the seeds of the Dutch variety alfalfa (Medicago sativa. L) were germinated commercially and four times between 2022 and 2024. The experiment was carried out in a completely randomized design (CRD) with two factors and three replicates in perforated dishes with dimensions of 28 cm in length and 19 cm in width (area 532 cm2) at a seed rate of kg / m2, resulting in an average seed weight of 53 g in one dish. The first experimental factor was the germination periods, while the second factor was soaking the seeds in solutions of varying amounts made from gibberellin and nano-compound fertilizer with the comparison treatment, which is soaking in water alone, as follows: Gibberellin 100 ppm and given the symbol GA 100ppm- Gibberellin 200 ppm and given the symbol 200ppm GA- --Nano fertilizer 20:20:20 NPK at a concentration of 2000 ppm and given the symbol NPK2000ppm. -Nano fertilizer 20:20:20 NPK at a concentration of 4000 ppm and given the symbol NPK4000ppm. This study showed that the Gibberellin 200ppm GA treatment was significantly superior to the rest of the other soaking treatments in the characteristics of the height of the incubator (3.66 cm), as well as the wet weight of both seedlings and roots and their dry weight, which were (497.88, 56.97g/dish-1), respectively, The fresh and dried root weights were considerably greater in the same treatment, at (400.86 and 43.86 g/dish-1), respectively. while the comparison treatment was superior. Morally, it was significantly superior to the rest of the other soaking treatments in the characteristic of protein in the medium was (18.29%). As for the times of hydroponic seed germination, there were no significant differences between the times in all the studied characteristics, which enhances the high degree of incubator isolation, the accuracy of implementing the experiment, and the accuracy of taking the results.

Effect of physical energy on germination and seedling vigor of alfalfa seeds (Medicago sativa L.)

Effect of physical energy on germination and seedling vigor of alfalfa seeds (Medicago sativa L.)

Rapid prediction and visualization of safe moisture content in alfalfa seeds based on multispectral imaging technology

Moisture significantly impacts seed sales, storage, and processing. Traditional moisture testing methods are often slow, labor-intensive, and inadequate for the rapid detection demands of modern agriculture, particularly for non-destructive testing of individual seeds. This study applied multispectral imaging to obtain morphological and spectral data from alfalfa seeds at six moisture levels (4 %, 8 %, 12 %, 16 %, 25 %, and 41 %). By integrating algorithms such as Support Vector Machines (SVM), Random Forests (RF), Linear Discriminant Analysis (LDA), Back Propagation Neural Network (BPNN), and normalized typical discriminant analysis (nCDA) algorithms, classification models were developed to distinguish between safe and unsafe moisture levels. The Results indicated that spectral data alone significantly improved model accuracy and prediction. nCDA visualizations effectively illustrated spatial moisture distribution, highlighting stark color differences between seeds in the safe moisture range (4 %, 8 %, 12 %) and those in the unsafe range (16 %, 25 %, 41 %). BPNN exhibited high model precision, achieving a recognition accuracy rate of 90.1 % for safe and unsafe moisture content. Key wavelengths identified by the Permutation method included 970, 880, 570, and 490 nm. Pearson correlation analysis showed a significant positive correlation between germination indicators and spectral data, which strengthened with longer seed storage. These findings confirm the potential of multispectral imaging for assessing the safe moisture content of alfalfa seeds, supporting the development of detection systems for evaluating moisture content in individual seeds. This advancement enables the rapid removal of high-moisture seeds, preventing deterioration during storage.

Effect of Ozone on Enterobacteriaceae Counts in Wheat Grain, Alfalfa, Radish, Broccoli Seeds and Sprouts

Abstract The sprouts of various seeds have garnered considerable regard amid consumers due to its verdure, crispiness, and germination potential, offering a cost-effective and straightforward process. The moist and warm conditions are very conducive to germination, fostering the growth of microorganisms and thereby raising food safety concerns. The aim of the study was to investigate the impact of ozone on Enterobacteriaceae counts by subjecting wheat (Triticum aestivum) grains, alfalfa (Medicago sativa), radish (Raphanus sativus) and broccoli (Brassica oleracea) seeds to ozone gas treatment. Additionally, the grain and seeds were rinsed with ozonated water during germination. Three different treatment methods were employed for seeds and grains. In the first method, samples were exposed to ozone gas at 20 ppm for 10 minutes, with subsequent rinsing every 12 hours during germination until the rinse water appeared visually clean, at a concentration of 2.0 mg L−1 of ozone. The second method involved rinsing with ozonated water at a concentration of 2.0 mg L−1 for durations of 20, 40, and 60 minutes. The third method - treated with ozone gas at 50 ppm and the exposure time of 1, 2, 3, 4 and 5 h. Untreated samples were used as controls. Evaluation of the samples revealed that Enterobacteriaceae was virtually undetectable in dry seeds. However, after 72 hours of germination in both the control and ozone-treated sprouts and following 7 days of storage at 4±2 °C, Enterobacteriaceae were found at an average of 7 to 8.1 log CFU g−1 in all samples, irrespective of the treatment method. Additionally, mould was observed only in wheat and radish sprouts during the assessment period. The study showed that there was no significant reduction in the number of mesophilic aerobic and facultative anaerobic microorganisms, moulds and Enterobacteriaceae then treated with ozone gas.

NaHS immersion alleviates the stress effect of chromium(III) on alfalfa seeds by affecting active oxygen metabolism

ABSTRACT Objective This study aimed to investigate the regulatory effects of exogenous hydrogen sulfide (H2S) on seed germination, seedling growth, and reactive oxygen species (ROS) homeostasis in alfalfa under chromium (Cr) ion (III) stress. Methods The effects of 0–4 mM Cr(III) on the germination and seedling growth of alfalfa were first assessed. Subsequently, following seed NaHS immersion, the influence of H2S on alfalfa seed germination and seedling growth under 2 mM Cr(III) stress was investigated, and the substance contents and enzyme activities associated with ROS metabolism were quantified. Results Compared to the control group, alfalfa plant germination was delayed under 2 mM Cr(III) stress for up to 48 h (p < 0.05). At 120 h, the total seedling length was approximately halved, and the root length was roughly one-third of the control. Treatment with 0.02–0.1 mM NaHS alleviated the delay in germination and root growth inhibition caused by 2 mM Cr(III) stress, resulting in an increased ratio of root length to hypocotyl length from 0.57 to 1 above. Additionally, immersion in 0.05 mM NaHS reduced hydrogen peroxide (H2O2) and oxygen-free radicals (O2 · –) levels (p < 0.05), boosted glutathione (GSH) levels (p < 0.05), and notably enhanced catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) activities (p < 0.05) compared to the 2 mM Cr(III) stress treatment group. Conclusion Seed immersion in NaHS mitigated the delay in germination and inhibition of root elongation under 2 mM Cr(III) stress. This effect is likely attributed to the regulation of intracellular ROS homeostasis and redox balance through enzymatic and non-enzymatic systems; thus, providing a potential mechanism for combating oxidative stress.

Role of malic acid in enhancing the efficiency of barley (Hordeum vulgare L.) and alfalfa (Medicago sativa L.) plants for phytoremediation of salt affected soil

Soil salinization is a growing global problem that influences plant growth and crop productivity. Most of the reclamation efforts in the past have focused on the installation of surface drainage systems. Other management approaches, such as excessive leaching and chemical amendments, have also been used on a limited scale to enhance the productivity of these soils. Phytoremediation can be cost-effective and environmentally sound technology.A laboratory experiment was carried out to study the role of malic acid which is low molecular weight organic acid (LMWOA) in enhancing the efficiency of barley and alfalfa plants for the phytoremediation of salt-affected soil. Seeds of barley and alfalfa were cultured in pots and irrigated with full strength Hoagland nutrient solution with three concentrations of seawater (SW) (10%, 20% and 30%) and a mixture of seawater with malic acid (MA) at 2, 4 and 6 mM l-1 (MA+SW), Hoagland solution was used as control. After twelve weeks, plants were harvested, and three types of soils (barley soil, alfalfa soil, and plant-free soil) were subjected to physical and chemical analysis for EC (electrical conductivity), TOC (total organic carbon), pH, potassium, sodium, and chloride ions. Results indicated a significant decrease was recorded in soil EC, pH, potassium, sodium, and chloride ions and a significant increase in soil TOC in barley and alfalfa soil compared with plant-free soil. Treatments with (MA+SW), especially at (2+10%) resulted in a significant increase in ions availability and phytoremediation activity in barley and alfalfa soils comparing with plant-free soil.

Alfalfa Spring Black Stem and Leaf Spot Disease Caused by Phoma medicaginis: Epidemic Occurrence and Impacts.

Alfalfa spring black stem and leaf spot disease (ASBS) is a cosmopolitan soil-borne and seed-borne disease caused by Phoma medicaginis, which adversely affects the yield, and nutritive value and can stimulate production of phyto-oestrogenic compounds at levels that may adversely affect ovulation rates in animals. This review summarizes the host range, damage, and symptoms of this disease, and general features of the infection cycle, epidemic occurrence, and disease management. ASBS has been reported from over 40 countries, and often causes severe yield loss. Under greenhouse conditions, reported yield loss was 31-82% for roots, 32-80% for leaves, 21% for stems and 26-28% for seedlings. In field conditions, the forage yield loss is up to 56%, indicating that a single-cut yield of 5302 kg/ha would be reduced to 2347 kg/ha. P. medicaginis can infect up to 50 species of plants, including the genera Medicago, Trifolium, Melilotus, and Vicia. ASBS is more severe during warm spring conditions before the first harvest than in hot summer and cooler winter conditions, and can infect alfalfa roots, stems, leaves, flowers, pods, and seeds, with leaf spot and/or black stem being the most typical symptoms. The primary infection is caused by the overwintering spores and mycelia in the soil, and on seeds and the cortex of dead and dry stems. The use of resistant cultivars is the most economical and effective strategy for the control of ASBS. Although biological control has been studied in the glasshouse and is promising, chemical control is the main control method in agriculture.

Transmission from seed to seedling and elimination of alfalfa viruses.

Viral diseases have become a vital factor limiting the development of the alfalfa (Medicago sativa) industry. Six viruses infecting alfalfa with a high incidence rate are Alfalfa mosaic virus (AMV), Medicago sativa alphapartitivirus 1 (MsAPV1), Medicago sativa alphapartitivirus 2 (MsAPV2), Medicago sativa deltapartitivirus 1 (MsDPV1), Medicago sativa amalgavirus 1 (MsAV1), and Cnidium vein yellowing virus 1 (CnVYV1). The purpose of this study was to develop preventive measures against these viruses by investigating their transmission through alfalfa seeds. In this study, we investigated the transmission rate of alfalfa viruses from seed to seedling by PCR, determined the location of viruses in seed by dissecting seed embryos and seed coat, tracked the changes of viruses in seedlings, and finally discover effective elimination measures for alfalfa viruses from 16 measures. Our results demonstrated that all these six viruses could be transmitted from alfalfa seeds to seedlings with the transmission rate ranging from 44.44% to 88.89%. For AMV, MsAPV2, and MsAV1, the viral load was significantly higher in the seed coats than in the seed embryos; however, it did not show significant differences between these two parts of the seeds for MsAPV1, MsDPV1, and CnVYV1. Dynamic accumulation analysis of AMV and MsAPV2 indicated that the viral load in plants increased continuously in the early growth stage, making it important to inactivate these viruses prior to their seed-to-seedling transmission. Sixteen treatments including physical, chemical, and combinations of physical and chemical measures were compared in terms of their elimination efficiency on AMV and MsAPV2 and impacts on seed germination. The results showed that soaking alfalfa seeds in sterile distilled water for 2h + 2% NaClO for 1h or 2% NaClO for 1h were more promisingly applicable because it could significantly reduce AMV and MsAPV2 particles in both seeds and seedlings. Our data revealed a route of virus transmission in alfalfa and shed light on the discovery of a highly efficient method for the management of alfalfa viral diseases.

Titanium dioxide nanoparticles (TiO2-NPs) effect on germination and morphological parameters in alfalfa, tomato, and pepper

Titanium dioxide nanoparticles are used in different processes, derived from this their presence in wastewater is common, concentrating in residual sludge. These residues are used as agricultural soil improvers, being a source of crop exposure. In this study, the effect of TiO2-NPs (450, 900, and 1800 mg L-1) on the germination of alfalfa, tomato, and pepper seeds was evaluated. The germination parameters were not different (p &gt; 0.05) except for, the seed vigor index in alfalfa and pepper, and the mean germination time in tomato. The germination index was below 80% in 450 mg L-1 in tomato, which showed moderate phytotoxicity. Morphological modifications with differences (p &lt; 0.05) were found in the three crops, mainly in the root. In tomatoes, the length of the main root, root hairs, and the width of the root tip were reduced but increased the width of the main root, piliferous zone, and the length of root hairs were. For alfalfa, root length and number of secondary roots augment. However, the stem, root tip width, and root villi were reduced. Finally, for pepper, the length, and width of the root and the piliferous zone were modified. Additionally, a concerning trend has been observed in the length of root hairs. TiO2-NPs affected germination and morphology in alfalfa, tomato and pepper seeds differently. Tomato was most negatively affected, with reduced root length and width. Alfalfa showed mixed effects, with positive impacts on some parameters but negatives on others. Pepper seeds responded positively overall, with improved germination and root length, despite some impacts on root morphology.

Alfalfa seeds potential in enhancing wheat flour nutritional composition, rheological properties and technological quality of resulting standard and sourdough bread

Alfalfa seeds potential in enhancing wheat flour nutritional composition, rheological properties and technological quality of resulting standard and sourdough bread

Non-Thermal Plasma (NTP) Treatment of Alfalfa Seeds in Different Voltage Conditions Leads to Both Positive and Inhibitory Outcomes Related to Sprout Growth and Nutraceutical Properties.

Non-thermal plasma (NTP) has proven to be a green method in the agricultural field for the stimulation of germination, growth, and production of nutraceutical compounds in some cases. However, the process is far from being fully understood and depends on the targeted plant species and the NTP used. In this work, we focus on the production of alfalfa sprouts from NTP-treated seeds under different voltage conditions. A flexible electrode configuration was used to produce the NTP, which can also be placed on packages for in-package treatments. The surface of the seeds was analyzed, indicating that the microstructure was strongly affected by NTP treatment. Biometric measurements evidenced the possibility of stimulating the sprout growth in some conditions by up to 50% compared to the sprouts obtained from untreated seeds. Biochemical traits for the sprouts obtained in different processing conditions were also studied, such as the concentrations of chlorophyll pigments, flavonoids and polyphenols, and antioxidant activity. Most NTP treatments led to inhibitory effects, proving the strong dependence between NTP treatment and targeted plant species.

Comparing native and non-native seed-isolated strains for drought resilience in maize (Zea mays L.)

Maize, a vital crop for human nutrition, livestock, and industrial development, faces increasingly severe climatic conditions that hinder its production capacity along with other strategic crops. Novel treatments based on microorganisms have demonstrated efficiency in enhancing plant development and responding to stress. The use of bacteria isolated from seeds is a novel approach for biotreatment, as recent studies point to a co-evolution process for their presence in seeds. This approach hypothesize a pre-adaptation to its host, which may lead to increased efficiency. However, several aspects of this approach remain understudied. In this study, we aimed to evaluate the potential of native maize seed microbiota in comparison to that isolated from other species to mitigate drought stress. For this we characterized seedborne microbiota of a common productive maize variety in Portugal, to use it as biotreatment in other two varieties (sensitive and resistant to drought), selecting the strain Pseudomonas fulva MB as the most promising candidate. Stenotrophomonas maltophilia MS-M1 strain, isolated from wild alfalfa seeds and previously characterized as a drought-tolerant enhancer, served as a non-native control strain. According to the data, both varieties of maize demonstrated enhanced vegetative growth when treated with both individual strains, as well with the consortium, with an increase in plant height of 5–7 % in full and medium irrigation, and 50–55 % when not irrigated. This trend was also observed in plant weight, which increased by 13–23 %, even under no irrigation. In addition, production in both varieties was positively impacted by these treatments, based on the amount of grain produced (by weight). The drought-sensitive variety experienced a 17 % increase under full irrigation, while the most tolerant variety experienced a 25–40 % increase. Under medium irrigation level, the increase was about 15 % in both varieties, while a 100 % and 140 % increase was observed in each variety, respectively, when no irrigation was applied. The results suggest that native strain the P. fulva MB was slightly more efficient treatment, as it outperformed the non-native strain in terms of productivity in both varieties. However, the differences were not solid enough along all parameters to consistently asses such difference. The consortium treatment only showed better performance under full or medium irrigation conditions for some production traits. These findings support the use of seed microbiota as very efficient biotreatments, suggesting than even non-native strains have a remarkable beneficial impact (interspecific), expanding the possible of use of this kind of bioinoculants. Further test are required to optimize the use of seed-isolated strains as better adapted or tailor-made solutions for agriculture.

Analysis and Identification of Differences in Volatile Components of Various Alfalfa Seeds Based on GC-IMS

Volatile components are one key factor in sample identification, differential analysis, quality control and origin traceability. In order to identify and analyze the differences in volatile substances in different alfalfa seeds, this study used gas chromatography–ion mobility spectrometry (GC-IMS), combined with the Gallery Plot plug-in, and PCA, PLS-DA and other analysis methods. In this way, a comprehensive analysis of volatile components in 10 alfalfa seeds, including aerospace varieties, was conducted. A comparative analysis of the characteristics of different sample compounds using topographic maps and fingerprints led to isolation of 48 kinds of 54 volatile compounds. Among them, esters (9 types), olefins (8 types), ketones (8 types), alcohols (6 types) and aldehydes (6 types) were found to be the most abundant volatile compounds in alfalfa seeds. At the same time, PCA and PLS-DA analysis models showed that esters, ketones and alcohols were the main volatiles causing the differences among alfalfa seeds. Among them, the content of various substances in the ZT2 and ZT3 aerospace varieties were higher than that of other varieties, while the types and contents of volatiles in ZT1, ZM2 and GN3 were relatively low. Therefore, in combination with the differences in maturity of each sample, the 10 varieties of alfalfa were finally divided into three categories, and the varieties of the same series were basically classified into one category. This provides a basis and convenience for future seed screening, identification, traceability and forage breeding.

Transcriptome Analysis Reveals the Vital Role of ABA Plays in Drought Tolerance of the ABA-Insensitive Alfalfa (Medicago sativa L.)

Drought stress severely affects alfalfa (Medicago sativa L.) growth and production. It is particularly important to analyze the key networks of drought in alfalfa through physiological and molecular levels. However, how to quickly screen drought-tolerant alfalfa germplasm and how to elucidate the molecular pathways of alfalfa responding to drought are less studied. In this study, based on our previous research, we further verified the association between the heritability of ABA sensitivity during seed germination and drought tolerance of plants and identified the key pathways of drought tolerance differences between ABA-sensitivity (S1-0) and -insensitivity (S1-50) plants via RNA-seq and analysis. The results showed that the sensitivity to ABA in alfalfa seeds can be inherited and that plants that are insensitive to ABA during germination show stronger drought tolerance. An analysis of the differentially expressed genes (DEGs) revealed that ABA biosynthesis and signaling, amino acid metabolism, LEA, and wax synthesis-related pathways may be the key pathways that can be used for drought tolerance improvement in alfalfa. DEGs such as NCED, PYR/PYL, and PP2C may contribute to drought tolerance in the S1-50 plant. The study further confirms that screening with ABA at the seed germination stage can select alfalfa lines with good drought tolerance, which provides a new theoretical basis for alfalfa drought tolerance breeding. The expression of the key genes of alfalfa in response to drought stress was also tested.

The fate of enterohemorrhagic Escherichia coli on alfalfa and fenugreek seeds and sprouts as affected by ascaroside #18 treatments

Alfalfa and fenugreek sprouts are healthy foods, but they are occasionally contaminated with bacterial pathogens and serve as vehicles for transmitting foodborne illnesses. This study examined the efficacy of ascaroside (ascr)#18 treatment for the control of enterohemorrhagic E. coli (EHEC) growth on sprouts. Commercial alfalfa and fenugreek seeds were decontaminated with 20,000 ppm of NaClO, and residual chlorine was neutralized with Dey-Engley broth. Decontaminated seeds were treated with 1 mM or 1 μM ascr#18, a plant immunity modulator, before being dried and mixed with sandy soil inoculated with E. coli F4546 or BAA-2326 at 104–105 CFU/g. The inoculated seeds were sprouted on 1 % water agar at 25 °C for 7 days in the dark. Seed or sprout samples were collected on days 0, 1, 3, 5, and 7 for enumeration of bacterial populations. Data was fit into the general linear model and analyzed using Fisher's least significant different test of the statistical analysis software. Treatment with ascr#18 significantly (P ≤ 0.05) reduced the cell population of EHEC on sprouts. The mean EHEC populations in the 1 mM or 1 μM treatment groups were 3.31 or 1.56 log CFU/g lower compared to the control groups. Besides treatment, sprout seed type and sprouting time were also significant independent variables influencing the growth of EHEC, according to the results of type III error analysis. However, EHEC strain type was not a significant independent variable. The study suggests that ascr#18 could be potentially used to control EHEC contamination and improve the microbial safety of sprouts.

Bacillus strain BX77: a potential biocontrol agent for use against foodborne pathogens in alfalfa sprouts.

Despite regulatory and technological measures, edible sprouts are still often involved in foodborne illness and are considered a high-risk food. The present study explored the potential of spore-forming Bacillus isolates to mitigate Salmonella and Escherichia coli contamination of alfalfa sprouts. Food-derived Bacillus strains were screened for antagonistic activity against S. enterica serovar Typhimurium SL1344 (STm) and enteropathogenic E. coli O55:H7. Over 4 days of sprouting, levels of STm and E. coli on contaminated seeds increased from 2.0 log CFU/g to 8.0 and 3.9 log CFU/g, respectively. Treatment of the contaminated seeds with the most active Bacillus isolate, strain BX77, at 7 log CFU/g seeds resulted in substantial reductions in the levels of STm (5.8 CFU/g) and E. coli (3.9 log CFU/g) in the sprouted seeds, compared to the control. Similarly, co-culturing STm and BX77 in sterilized sprout extract at the same ratio resulted in growth inhibition and killed the Salmonella. Confocal-microscopy experiments using seeds supplemented with mCherry-tagged Salmonella revealed massive colonization of the seed coat and the root tip of 4-day-old sprouted seeds. In contrast, very few Salmonella cells were observed in sprouted seeds grown with BX77. Ca-hypochlorite disinfection of seeds contaminated with a relatively high concentration of Salmonella (5.0 log CFU/g) or treated with BX77 revealed a mild inhibitory effect. However, disinfection followed by the addition of BX77 had a synergistic effect, with a substantial reduction in Salmonella counts (7.8 log CFU/g) as compared to untreated seeds. These results suggest that a combination of chemical and biological treatments warrants further study, toward its potential application as a multi-hurdle strategy to mitigate Salmonella contamination of sprouted alfalfa seeds.

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-1β), 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.