Search

Introduction: Alfalfa (Medicago sativa L.) is a globally important legume crop with high nutritional and ecological value. Drought poses a serious threat to alfalfa acreage and yields. Spermine (Spm) has been shown to protect plants from drought damage. The aim of this study was to clarify the mechanism of exogenous Spm to improve drought resistance of alfalfa. Methods: In this study, we root applied 0.1, 0.5, and 1mM Spm to Gannong No. 3 (G3) alfalfa under drought stress, and then determined their physiological and metabolic changes. Results: The results showed that exogenous Spm increased chlorophyll content, chlorophyll fluorescence parameters and gas exchange parameters, enhanced antioxidant enzymes activity, improved ascorbic acid-glutathione (AsA-GSH) cycle, increased osmoregulatory substances content, reduced hydrogen peroxide and superoxide anion levels, and inhibited malondialdehyde accumulation in alfalfa under drought stress, thereby increasing plant height and leaf relative water content and enhancing drought tolerance of alfalfa. The redundancy analysis of the above physiological indicators showed that the addition of the optimal Spm to improve drought tolerance of alfalfa under drought stress was mainly achieved by increasing catalase activity and improving the ASA-GSH cycle. In addition, metabolomics analysis revealed that exogenous Spm increased the content of oxobutanedioic acid, citric acid, fumaric acid and malic acid to enhance the tricarboxylic acid cycle. Meanwhile, exogenous Spm increased endogenous Spm and proline (Pro) content to resist drought stress by enhancing Spm and Pro metabolism. Moreover, exogenous Spm increased the accumulation of the signaling substance abscisic acid. Discussion: In conclusion, exogenous Spm enhanced drought resistance of alfalfa leaves under drought stress. [ABSTRACT FROM AUTHOR]

Pollination stands out as the paramount ecosystem service facilitated by insect pollinators, providing benefits to over three quarters of the world’s major crops. The conservation and effective management of honeybees constitute integral components for ensuring the sustainable productivity of cross-pollinated crops. The present study aimed to document the potential pollinators of lucerne in the Bundelkhand region in India, which is known for its subtropical climatic conditions. Further, the foraging behaviour, including foraging rate and speed, was meticulously documented. A total of 13 insect species from seven families across three insect orders were observed visiting lucerne flowers. Hymenopterans comprised the majority of the floral visitors (73.48%), followed by Lepidoptera (26.51%) during the study period. Among them, Apis dorsata (Hymenoptera: Apidae) emerged as the most predominant floral visitor, exhibiting peak activity at 12:00 h with a density of 7.85 bees/m2/10 min. Surprisingly, among the major floral visitors, A. florea has the highest mean foraging rate (number of flowers visited/minute/forager), i.e., 11.56, 11.59 and 10.79 bees/flowers/minute during the initial flowering stage, peak flowering stage and late flowering stage, respectively. However, the foraging speed (time spent by the forager on each flower) was longest in A. dorsata, i.e., 5.05, 3.78 and 5.41 s/flower during initial, peak and late flowering stages, respectively. The findings emphasize the importance of enhancing honeybee activity to boost lucerne seed yield. Further, the outcomes of the study can be leveraged to optimize honeybee colony management, scheduling field operations such as pesticide application and weeding during periods of minimal honeybee activity. [ABSTRACT FROM AUTHOR]

Background: Drought stress is a major limiting factor that affects forage yields, and understanding the drought resistance mechanism of plants is crucial for improving crop yields in arid areas. Alfalfa (Medicago sativa L.) is the most important legume plant, mainly planted in arid and semi-arid areas. However, the adaptability of alfalfa to drought stress and its physiological and molecular mechanisms of drought resistance remains unclear. Results: In this study, we analyzed the physiological and transcriptome responses of alfalfa cultivars with different drought resistances (drought-sensitive Gannong No. 3 (G3), drought-resistant Gannong No. 8 (G8), and strong drought-resistant Longdong (LD)) under drought stress at 0, 6, 12, and 24 h. LD had higher catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities and a higher soluble protein content, lower malondialdehyde (MDA) content, a lower O2·− production rate, and a lower H2O2 content than G8 and G3 (P < 0.05). The functional enrichment analysis, temporal expression pattern analysis, and weighted gene co-expression network analysis (WGCNA) of the differentially expressed genes (DEGs) showed phenylpropanoid biosynthesis, flavonoid biosynthesis, starch and sucrose metabolism, glycolysis/gluconeogenesis, glutathione metabolism, and biosynthesis of amino acid responses to drought stress in alfalfa. The differential expression of genes during phenylpropanoid biosynthesis, starch and sucrose metabolism, and the glutathione metabolism pathway was further studied, and it was speculated that PAL, COMT, 4CL, CCR, CAD, HXK, INV, SUS, WAXY, AGP, GST, and APX1 played important roles in the alfalfa drought stress response. Conclusions: The aim of this study was to enhance alfalfa drought resistance by overexpressing positively regulated genes and knocking out negatively regulated genes, providing genetic resources for the subsequent molecular-assisted breeding of drought-resistant alfalfa crops. [ABSTRACT FROM AUTHOR]

The cultivation of alfalfa is crucial for farmers as it is an excellent forage crop with a high nitrogen-fixing capacity, making it indispensable in crop rotations. Breeding programs face challenges in advancing more rapidly in genetic diversity to achieve a higher heterosis effect and, consequently, greater yield. In this study, we used 30 alfalfa varieties, which were used for molecular analyses by 5 ISSR primers and 13 RAPD primers. The results obtained highlighted the greater efficiency of ISSR primers in identifying genetic diversity. On the other hand, the simultaneous use of ISSR + RAPD allowed for clearer clustering of varieties that enabled more efficiently distinguishing the genetic diversity. The most efficient ISSR primer, A17, generated 31 polymorphic bands, while the most efficient RAPD primer, L-07, generated only 21 bands. Varieties such as "Pastoral" and "F1413-02" exhibited low similarity coefficients (0.39), suggesting their potential for enhancing genetic variability through crossbreeding, thereby increasing the potential of achieving a greater heterosis effect. Conversely, varieties with high similarity coefficients, such as "Cristal" and "Viking" (0.81) are less suited for this purpose. The correlation between specific markers highlights that using both ISSR and RAPD markers together offers a clear understanding of genetic diversity in alfalfa, aiding in more effective selection for crossbreeding in breeding programs. [ABSTRACT FROM AUTHOR]

In order to study the suitable drip head flow, buried depth and irrigation quota of alfalfa under subsurface drip irrigation, field experiments were carried out to study the single factor effects of key technical parameters of subsurface drip irrigation on alfalfa root system and yield by setting three drip irrigation areas with drip head flow (1.38, 2.00 and 3.00 L/h), three buried depths of drip irrigation area (10, 20 and 30 cm) and three subsurface drip irrigation water quota (15.0, 22.5 and 30.0 mm). The results show that when the drip flow rate is 2.00 L/h, the growth of alfalfa root system is more favorable and the buried depth of drip irrigation has a significant effect on lateral roots of alfalfa. Irrigation quota has significant effects on specific root surface area, root tip number, lateral root occurrence number and lateral root position of alfalfa. The yield of alfalfa treated with small drip flow is generally higher than that treated with large drip flow. The buried depth of drip irrigation area has no significant effect on alfalfa yield. In a certain range, the yield of alfalfa is increased gradually with the increase of irrigation quota. Considering the factors of water saving and yield, it is suggested that the key technical parameters of alfalfa subsurface drip irrigation in Etoke Qianqi area of Ordos are as follows: The drip head flow is 2.00 L/h, the buried depth of drip irrigation area is 20 cm, and the irrigation quota is 22.5 mm. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) can fix N naturally within soils, which makes alfalfa cultivation useful for enhancing soil fertility while minimizing environmental impacts from pesticides, fertilizers, and soil pollution. To assess the influence of alfalfa cropping on degraded black soil, we determined the nutrient stoichiometry of the soil and soil microbial biomass under four corn cultivation systems at the Harbin Corn Demonstration Base (Heilongjiang, China), which is located in Wujia (126°23′ E, 45°31′ N), Shuangcheng district, Harbin, Heilongjiang Province. The cultivation systems included continuous corn cultivation for more than 30 years (CK), 2 years of alfalfa–corn rotation (AC), three years of alfalfa cropping (TA), and four years of alfalfa cropping (FA). Overall, AC, TA, and FA treatment increased the soil pH, reduced the soil salinity, and increased the organic matter content of the 0–15 cm soil layer. TA and FA presented soil nutrient levels comparable to those of degraded cornfields that were fertilized annually. The TA and FA treatments increased the soil available N:P, soil N:P, and soil C:P ratios. Moreover, TA significantly increased the soil microbial biomass P (SMBP) in the 0–15 cm (surface) soil layer and reduced the soil microbial biomass C (SMBC):SMBP ratio. AC, TA, and FA increased the storage and mineralization rates of soil N and alleviated the microbial P limitations in degraded black soil farmland. Compared with FA, TA resulted in greater improvements in the quality of degraded black soil farmland. The ability of alfalfa to enhance soil fertility makes an important component of sustainable agricultural practices aimed at rehabilitating degraded soils. [ABSTRACT FROM AUTHOR]

Gravid culicine mosquitoes rely on olfactory cues for selecting breeding sites containing organic detritus. While this capacity of the mosquitoes is used for surveillance and control, the current methodology is unwieldy, unreliable and expensive in time and labour. This study evaluated the dose-dependent attraction and oviposition response of gravid Culex quinquefasciatus to alfalfa infusions. Through combined chemical and electrophysiological analyses, bioactive volatile organic compounds (VOCs) in the headspace of alfalfa infusions, eliciting attraction, were identified. While phenolic and indolic compounds were the most abundant bioactive VOCs, additional VOCs, including a monoterpene, were required to elicit a significant behavioural response to the synthetic odour blend of alfalfa infusions. Comparative analysis with the commercially available mosquito oviposition pheromone (MOP) was also conducted demonstrating that this standardised synthetic alfalfa infusion odour blend offers a promising lure for targeted surveillance and control of Culex mosquitoes, which may contribute to disease prevention and public health protection. [ABSTRACT FROM AUTHOR]

JASMONATE ZIM domain (JAZ) proteins, inhibitors of the jasmonic acid (JA) signaling pathway, are identified in different plants, such as rice and Arabidopsis. These proteins are crucial for growth, development, and abiotic stress responses. However, limited information is available regarding the JAZ family in alfalfa. This study identified 11 JAZ genes (MsJAZs) in the "Zhongmu No.1" reference genome of alfalfa. The physical and chemical properties, chromosome localization, phylogenetic relationships, gene structure, cis-acting elements, and collinearity of the 11 MsJAZ genes were subsequently analyzed. Tissue-specific analysis revealed distinct functions of different MsJAZ genes in growth and development. The expression patterns of MsJAZ genes under salt stress conditions were validated using qRT-PCR. All MsJAZ genes responded to salt stress, with varying levels of upregulation over time, highlighting their role in stress responses. Furthermore, heterogeneous expression of MsJAZ1 in Arabidopsis resulted in significantly lower seed germination and survival rates in OE-2 and OE-4 compared to the WT under 150 mM NaCl treatment. This study establishes a foundation for further exploration of the function of the JAZ family and provides significant insights into the genetic improvement of alfalfa. [ABSTRACT FROM AUTHOR]

Alfalfa species Medicago sativa L. (MS) and Medicago falcata L. (MF), globally prominent perennial leguminous forages, hold substantial economic value. However, our comprehension of the molecular mechanisms governing their resistance to cold stress remains limited. To address this knowledge gap, we scrutinized and compared MS and MF cold-stress responses at the molecular level following 24 h and 120 h low-temperature exposure (4 °C). Our study revealed that MF had superior physiological resilience to cold stress compared with MS, and its morphology was healthier under cold stress, and its malondialdehyde content and superoxide dismutase activity increased, first, and then decreased, while the soluble sugar content continued to accumulate. Transcriptome analysis showed that after 120 h of exposure, there were different gene-expression patterns between MS and MF, including 1274 and 2983 genes that were continuously up-regulated, respectively, and a total of 923 genes were included, including star cold-resistant genes such as ICE1 and SIP1. Gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed numerous inter-species differences in sustained cold-stress responses. Notably, MS-exclusive genes included a single transcription factor (TF) gene and several genes associated with a single DNA repair-related pathway, whereas MF-exclusive genes comprised nine TF genes and genes associated with 14 pathways. Both species exhibited high-level expression of genes encoding TFs belonging to AP2-EREBP, ARR-B, and bHLH TF families, indicating their potential roles in sustaining cold resistance in alfalfa-related species. These findings provide insights into the molecular mechanisms governing cold-stress responses in MS and MF, which could inform breeding programs aimed at enhancing cold-stress resistance in alfalfa cultivars. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) ‐ grass or alfalfa ‐ sainfoin (Onobrychis viciifolia Scop.) mixtures are commonly used for pastures in western Canada because of their high forage quality and their low risk of causing frothy bloat in grazing animals. However, the proportion of these two legumes declines in mixed forage stands over time. The objective of this study was to evaluate the effectiveness of selecting superior genotypes from clonally propagated alfalfa or sainfoin under plant competition in different growth environments. For this study, around 100 genotypes of each legume were cloned and transplanted into meadow bromegrass (Bromus riparius Relm.) or alfalfa swards at Saskatoon, SK and Lethbridge, AB, Canada in 2017. Genotype‐environment (G x E) interactions of alfalfa and sainfoin genotypes were analysed by an additive main‐effects and multiplicative interaction (AMMI) model. Significant variations in plant height, spring vigour and total dry matter yield (TDM) were observed for both species. Among the measured traits, plant height was a more highly heritable trait (H2 =.16 for alfalfa and H2 =.18 for sainfoin), while TDM was the least heritable (H2 =.08 for alfalfa and H2 =.04 for sainfoin). In the AMMI Analysis of variance for TDM, the genotype explained <12% of the variation for both species, suggesting a direct selection of yield would result in a low genetic gain. The biplot mean performance (Y) x weighted average of absolute scores from the singular value decomposition of the matrix of BLUPs stability index (WAASB) identified several promising genotypes with superior performance and stability across different environments that were selected for producing synthetic lines. However, the synthetic lines of both alfalfa and sainfoin did not consistently exhibit superior performance in mixtures compared to their respective check cultivars. [ABSTRACT FROM AUTHOR]

This study investigates the impact of phosphorus fertilization on the yield and quality of various alfalfa (Medicago sativa L.) varieties grown in light chestnut soils. Conducted over a three-year period from 2013 to 2015, the research was carried out in the Karasay district of the Almaty region under irrigated conditions. The experiment included six alfalfa varieties: NS Alfa, VS Banat, Mediana, Nera, Niagara, and Kokoray. Four phosphorus treatments were applied: control (no phosphorus), 60 kg/ha (P60), 90 kg/ha (P90), and 120 kg/ha (P120), using double superphosphate as the phosphorus source. The results demonstrated that phosphorus fertilization significantly enhanced both the yield and quality of alfalfa. Across all varieties, the highest yield was observed with the application of 120 kg/ha phosphorus. For instance, NS Alfa's yield increased from 283.3 c/ha in the control to 349.7 c/ha with P120, reflecting a 23% increase. Similarly, VS Banat and Mediana exhibited yield increases of 23% and 25%, respectively, at the highest phosphorus level. The study also revealed improvements in the nutritional quality of alfalfa hay. Crude protein content increased from 20.3% to 22.0% in NS Alfa, while digestible protein content rose from 11.20% to 12.40%. Other quality parameters, including fat and carotene content, also improved significantly with higher phosphorus levels. Moreover, the availability of nitrate nitrogen and mobile phosphorus in the soil increased progressively with higher phosphorus application rates, contributing to better nutrient uptake and overall plant health. This research underscores the importance of phosphorus fertilization in maximizing alfalfa yield and quality. The findings suggest that the optimal phosphorus application rate for enhancing alfalfa production in light chestnut soils is 120 kg/ha, providing valuable insights for sustainable agricultural practices in similar agro-ecological zones. [ABSTRACT FROM AUTHOR]

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

Three bacteria strains of Pseudomonas sp. resistant to heavy metals were isolated from chemically contaminated soil. An analysis of the nucleotide sequences of the 16S rRNA and rpoD genes identified the Pseudomonas sp. 17 НМ strain as Pseudomonas capeferrum, and the strains of Pseudomonas sp. 65 НМ and 67 НМ were most closely related to the type strains of Pseudomonas silesiensis and Pseudomonas umsongensis, respectively. It was shown that the strains of Pseudomonas sp. 17 НМ, 65 НМ, and 67 НМ were characterized by different levels of resistance to heavy metals, with the maximum tolerable concentration (MTC) of zinc being 1 mМ for all strains, that of cadmium, 1, 1.5, and 1 mМ; of lead, 5, 5, and 4 mМ; and of nickel, 7, 9, and 7 mМ, respectively. All Pseudomonas strains can form biofilms and have the properties of PGPR bacteria. Treatment of alfalfa seeds (Medicago sativa L.) with the strains of Pseudomonas sp. 17 НМ, 65 НМ, and 67 НМ under cadmium stress increased the dry weight of alfalfa seedlings up to 40% and the content of chlorophyll a and b in the leaves by 25–33% relative to the control. [ABSTRACT FROM AUTHOR]

The wheat flour-alfalfa blends containing 5 and 10 g/100 g of non-germinated and germinated alfalfa flour were examined in terms of nutritional composition (protein, lipid, starch, total dietary fibre), mineral profile, total phenolic content and antioxidant activity (ABTS, FRAP), and Mixolab rheological properties while the corresponding standard and sourdough breads were evaluated regarding physical and textural characteristics (specific volume, hardness, cohesiveness, springiness, gumminess, chewiness). The alfalfa flour inclusion resulted in increased protein (11.64–14.87%) and total dietary fibre (1.65–5.82%) contents in all blends, regardless of the alfalfa flour type but in a higher extent with 10 g/100 g inclusion level. Blends were characterised by improved mineral profile, total phenolic content (37.57–162.35 mg GAE/100 g) and antioxidant properties (ABTS: 4.37–232.79 µmol TE/100 g, FRAP: 138.05–539.16 µmol TE/100 g) particularly when non-germinated alfalfa flour in quantity of 10 g/100 g was used. Dough rheological properties were not significantly affected by the alfalfa flour type, whilst higher alfalfa flour inclusion level prolonged dough development time. Increased specific volume (2.63–3.62 cm3/g) with decreased crumb hardness (3.82–1.68 N) was recorded upon alfalfa flour incorporation and was more pronounced for sourdough bread. Overall nutrient composition of blends and technological features of the resulting breads were enhanced by alfalfa flour addition. [ABSTRACT FROM AUTHOR]

Conditioning is an important step in harvesting alfalfa hay, as squeezing and bending alfalfa stems can break down the stem fibers and accelerate the drying rate of alfalfa. The quality of alfalfa hay is directly affected by the conditioning effect. The finite element method (FEM) can quantitatively analyze the interaction relationship between alfalfa and conditioning rollers, which is of great significance for improving conditioning effects and optimizing conditioning systems. The accuracy of material engineering parameters directly affects the simulation results. Due to the small diameter and thin stem wall of alfalfa, some of its material parameters are difficult to measure or have low measurement accuracy. Based on this background, this study proposed a method for calibrating the finite element parameters of thin-walled plant stems. By conducting radial tensile, shear, bending, and radial compression tests on alfalfa stems and combining with the constitutive relationship of the material, the range of engineering parameters for the stems was preliminarily obtained. By conducting a Plackett–Burman experiment, the parameters that affect the maximum shearing force of stems were determined, including Poisson's ratio in the isotropic plane, radial elastic modulus, and the sliding friction coefficient between the alfalfa stem and steel plate. By conducting the steepest ascent experiment and Box–Behnken experiment, the optimal values of Poisson's ratio, radial elastic modulus, and sliding friction coefficient were obtained to be 0.42, 28.66 MPa, and 0.60, respectively. Finally, the double-shear experiment, radial compression experiment, and conditioning experiment were used to evaluate the accuracy of the parameters. The results showed that the average relative error between the maximum shear and the measured value was 0.88%, and the average relative error between the maximum radial contact force and the measured value was 2.13%. In the conditioning experiment, the load curve showed the same trend as the measured curve, and the simulation results could demonstrate the stress process and failure mode of alfalfa stems. The modeling and calibration method can effectively predict the stress and failure of alfalfa during conditioning. [ABSTRACT FROM AUTHOR]

Introduction: Intercropping and soil properties both affect soil diazotrophic communities. However, the specific effects that alfalfa-maize intercropping has on diazotrophic networks and community diversity under different soil properties remain unclear. Methods: In this study, we investigated the soil diazotrophic communities of two crop systems, alfalfa monoculture (AA) and alfalfa-maize intercropping (A/M), in two sites with similar climates but different soil properties (poor vs. average). Results and discussion: The diazotrophic network complexity and community diversity were higher at the site with poor soil than at the site with average soil (p < 0.05). Community structure also varied significantly between the sites with poor and average soil (p < 0.05). This divergence was mainly due to the differences in soil nitrogen, phosphorus, and organic carbon contents between the two sites. At the site with poor soil, the A/M system had lower diazotrophic diversity, lower network complexity and greater competition between diazotrophs than the AA system (p < 0.05) because intercropping intensified the soil phosphorus limitation under poor soil conditions. However, in the average soil, it was the A/Msystemthat had an altered diazotrophic structure, with an increased abundance of 11 bacterial genera and a decreased abundance of three bacterial genera (p < 0.05). Conclusion: Our results indicated that the effects of alfalfa-maize intercropping on diazotrophic communities were soil property-dependent. [ABSTRACT FROM AUTHOR]

Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Sprouted seeds and microgreens are emerging as innovative specialty raw salad crops, valued for their health-promoting properties. However, the risk of foodborne illnesses associated with microbial contamination in microgreens underscores the need for effective sanitization in their production. This study explores using hydrolates, by-products of essential oil distillation from six plants, as natural sanitizers in alfalfa microgreen production. The research investigates their impact on seed germination, antimicrobial activity, and sensory attributes. Results revealed that oregano, fennel, lavender, and lemon catmint hydrolates decrease alfalfa seed germination, while peppermint and hop hydrolates have no significant impact. Peppermint and fennel hydrolates demonstrate notable efficacy in antimicrobial testing. Sensory analysis indicates differences in odour and flavour, with peppermint, oregano, and lavender receiving high scores. According to the results, peppermint hydrolate can be considered a favourable option for alfalfa micro sprout production, contributing to sustainable and organic approaches in urban agriculture and underlining the importance of natural sanitizers for food safety. Highlights: Micro sprouts are prevalent raw salad crops with functional properties. Alfalfa sprouting was done in water or six selected hydrolates during 10 days. Peppermint, oregano, fennel, hop, lavender and lemon catmint hydrolates were used. Peppermint hydrolate showed the highest antimicrobial and sensory potential. [ABSTRACT FROM AUTHOR]

Background: Medicago sativa, often referred to as the "king of forage", is prized for its high content of protein, minerals, carbohydrates, and digestible nutrients. However, various abiotic stresses can hinder its growth and development, ultimately resulting in reduced yield and quality, including water deficiency, high salinity, and low temperature. The ethylene-insensitive 3 (EIN3)/ethylene-insensitive 3-like (EIL) transcription factors are key regulators in the ethylene signaling pathway in plants, playing crucial roles in development and in the response to abiotic stresses. Research on the EIN3/EIL gene family has been reported for several species, but minimal information is available for M. sativa. Results: In this study, we identified 10 MsEIN3/EIL genes from the M. sativa genome (cv. Zhongmu No.1), which were classified into three clades based on phylogenetic analysis. The conserved structural domains of the MsEIN3/EIL genes include motifs 1, 2, 3, 4, and 9. Gene duplication analyses suggest that segmental duplication (SD) has played a significant role in the expansion of the MsEIN3/EIL gene family throughout evolution. Analysis of the cis-acting elements in the promoters of MsEIN3/EIL genes indicates their potential to respond to various hormones and environmental stresses. We conducted a further analysis of the tissue-specific expression of the MsEIN3/EIL genes and assessed the gene expression profiles of MsEIN3/EIL under various stresses using transcriptome data, including cold, drought, salt and abscisic acid treatments. The results showed that MsEIL1, MsEIL4, and MsEIL5 may act as positive regulatory factors involved in M. sativa's response to abiotic stress, providing important genetic resources for molecular design breeding. Conclusion: This study investigated MsEIN3/EIL genes in M. sativa and identified three candidate transcription factors involved in the regulation of abiotic stresses. These findings will offer valuable insights into uncovering the molecular mechanisms underlying various stress responses in M. sativa. [ABSTRACT FROM AUTHOR]

Alfalfa biomass can be fractionated into leaf and stem components. Leaves comprise a protein-rich and highly digestible portion of biomass for ruminant animals, while stems constitute a high fiber and less digestible fraction, representing 50 to 70% of the biomass. However, little attention has focused on stem-related traits, which are a key aspect in improving the nutritional value and intake potential of alfalfa. This study aimed to identify molecular markers associated with four morphological traits in a panel of five populations of alfalfa generated over two cycles of divergent selection based on 16-h and 96-h in vitro neutral detergent fiber digestibility in stems. Phenotypic traits of stem color, presence of stem pith cells, winter standability, and winter injury were modeled using univariate and multivariate spatial mixed linear models (MLM), and the predicted values were used as response variables in genome-wide association studies (GWAS). The alfalfa panel was genotyped using a 3K DArTag SNP markers for the evaluation of the genetic structure and GWAS. Principal component and population structure analyses revealed differentiations between populations selected for high- and low-digestibility. Thirteen molecular markers were significantly associated with stem traits using either univariate or multivariate MLM. Additionally, support vector machine (SVM) and random forest (RF) algorithms were implemented to determine marker importance scores for stem traits and validate the GWAS results. The top-ranked markers from SVM and RF aligned with GWAS findings for solid stem pith, winter standability, and winter injury. Additionally, SVM identified additional markers with high variable importance for solid stem pith and winter injury. Most molecular markers were located in coding regions. These markers can facilitate marker-assisted selection to expedite breeding programs to increase winter hardiness or stem palatability. [ABSTRACT FROM AUTHOR]

MicroRNA (miRNA) is a type of endogenous non-coding small RNA, which is abundant in living organisms. miRNAs play an important role in regulating gene expression and myriad cellular processes by binding to target messenger RNAs through complementary base pairing, and cross-species regulation mammalian cells by plant-derived xeno-miRNAs has been described. Here, we examined the miRNA species in two alfalfa (Medicago sativa, lucerne) cultivars commonly grown in Ningxia, China: cv. Zhongmu 1 and cv. Xinyan 52. Both cultivars have good salt and drought resistance. We found that the miRNA profiles were similar between the cultivars, with a slightly higher number of miRNAs present in the newer cv. Xinyan 52, which may contribute to its improved salt and drought tolerance. miRNAs were stable during drying, and some miRNAs were increased in dry versus fresh alfalfa, suggesting some miRNAs may be upregulated during drying. Alfalfa-derived miRNAs could be detected in exosomes from serum and whey collected from dairy cows, confirming the ability of the exogenous miRNAs (xeno-miRNAs) to enter the circulation and reach the mammary epithelium. In vitro studies confirmed that overexpression of mtr-miR156a could downregulate expression of Phosphatase 2 Regulatory Subunit B'gamma (PPP2R5D) and Phosphoinositide-3-kinase Regulatory Subunit 2 (PIK3R2). Overexpression of mtr-miR156a also modulated PI3K–AKT–mTOR signaling as well as the casein content of milk produced by bovine mammary epithelial cells. Based on the known roles of PPP2R5D and PIK3R2 in regulating the PI3K–AKT–mTOR pathway as well as the effect of PI3K–AKT–mTOR on milk protein content, our findings implicate alfalfa-derived miR156a as a new cross-species regulator of milk quality in dairy cows. [ABSTRACT FROM AUTHOR]

Estimating pasture biomass has emerged as a promising avenue to assist farmers in identifying the best cutting times for maximizing biomass yield using satellite data. This study aims to develop an innovative framework integrating field and satellite data to estimate aboveground biomass in alfalfa (Medicago sativa L.) at farm scale. For this purpose, samples were collected throughout the 2022 growing season on different mowing dates at three fields in Kansas, USA. The satellite data employed comprised four sources: Sentinel-2, PlanetScope, Planet Fusion, and Biomass Proxy. A grid of hyperparameters was created to establish different combinations and select the best coefficients. The permutation feature importance technique revealed that the Planet's PlanetScope near-infrared (NIR) band and the Biomass Proxy product were the predictive features with the highest contribution to the biomass prediction model's. A Bayesian Additive Regression Tree (BART) was applied to explore its ability to build a predictive model. Its performance was assessed via statistical metrics (r2: 0.61; RMSE: 0.29 kg.m−2). Additionally, uncertainty quantifications were proposed with this framework to assess the range of error in the predictions. In conclusion, this integration in a nonparametric approach achieved a useful predicting tool with the potential to optimize farmers' management decisions. [ABSTRACT FROM AUTHOR]

Although previous studies have demonstrated that methane (CH4) can mitigate the toxicity of cadmium (Cd) in alfalfa seedlings, the CH4-rich water used in these studies may create hypoxic conditions, potentially influencing the experimental outcomes. Therefore, this study aimed to investigate whether CH4 can reduce Cd toxicity in alfalfa seedlings without the interference of hypoxia and to analyze its underlying mechanisms. Here, it was observed that supplementing oxygen with saturated CH4-rich water can significantly alleviate the inhibition of 75 μM CdCl2 on the growth of alfalfa (Medicago sativa L.) seedlings. Less Cd accumulation was also observed in both root and shoot parts, which could be explained by the CH4-altered cell wall components in alfalfa seedling roots, including covalent and ionic soluble pectin, and the degree of demethylation in pectin, thus enabling a higher proportion of Cd binding to the cell walls and reducing the entry of Cd into the cells. The above actions of CH4 were accompanied by an increase in hydrogen peroxide (H2O2) content and NADPH oxidase activity, which could be blocked by the addition of the NADPH oxidase inhibitor diphenylene iodonium (DPI). Taken together, these results implied that exogenously applied CH4 could alleviate Cd toxicity in alfalfa seedlings by enhancing Cd chelation onto the root cell walls, which might be closely associated with NADPH oxidase-dependent H2O2 signals. These findings could provide insight into the mechanism through which CH4 alleviates Cd toxicity in alfalfa plants. [ABSTRACT FROM AUTHOR]

The growth and biological decline of alfalfa may be linked to the rhizosphere microbiome. However, plant–microbe interactions in the rhizosphere of alfalfa and associated microbial community variations with stand age remain elusive. This study explored the successional pattern of rhizosphere microbial communities across different aged alfalfa stands and its relationship with alfalfa decline. Rhizosphere soils were collected from 2- and 6-year-old alfalfa stands. Control soils were collected from interspaces between alfalfa plants in the same stands. Soil bacterial and fungal communities were characterized by 16S and ITS rRNA gene sequencing, respectively. Specific microbial taxa colonized the rhizosphere soils, but not the control soils. The rhizosphere-specific taxa mainly included potentially beneficial genera (e.g., Dechloromonas, Verrucomicrobium) in the young stand and harmful genera (e.g., Peziza, Campylocarpon) in the old stand. Alfalfa roots regulated soil microbial communities by selective promotion or inhibition of distinct taxa. The majority of time-enriched taxa were reported as harmful fungi, whose relative abundances were negatively correlated with plant traits. Time-depleted taxa were mostly known as beneficial bacteria, which had relative abundances positively correlated with plant traits. The relative abundances of functional bacterial genes associated with vancomycin biosynthesis, zeatin biosynthesis, and amino acid metabolism trended lower in rhizosphere soils from the old stand. An upward trend was observed for fungal pathogens and wood saprotrophs with increasing stand age. The results suggest that root activity drives the negative succession of rhizosphere microbial communities during alfalfa decline in old stands. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal fungi (AMF) are universally distributed in soils, including saline soils, and can form mycorrhizal symbiosis with the vast majority of higher plants. This symbiosis can reduce soil salinity and influence plant growth and development by improving nutrient uptake, increasing plant antioxidant enzyme activity, and regulating hormone levels. In this study, rhizosphere soil from eight plants in the Songnen saline-alkaline grassland was used to isolate, characterize, and screen the indigenous advantageous AMF. The promoting effect of AMF on alfalfa (Medicago sativa L.) under salt treatment was also investigated. The findings showed that 40 species of AMF in six genera were identified by highthroughput sequencing. Glomus mosseae (G.m) and Glomus etunicatum (G.e) are the dominant species in saline ecosystems of northern China. Alfalfa inoculated with Glomus mosseae and Glomus etunicatum under different salt concentrations could be infested and form a symbiotic system. The mycorrhizal colonization rate and mycorrhizal dependence of G.m inoculation were significantly higher than those of G.e inoculation. With increasing salt concentration, inoculation increased alfalfa plant height, fresh weight, chlorophyll content, proline (Pro), soluble sugar (SS), soluble protein (SP), peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) activity while decreasing the malondialdehyde (MDA) content and superoxide anion production rate. The results highlight that inoculation with G.m and G.e effectively alleviated salinity stress, with G.m inoculation having a significant influence on salt resistance in alfalfa. AMF might play a key role in alfalfa growth and survival under harsh salt conditions. [ABSTRACT FROM AUTHOR]

Background: Alfalfa (Medicago sativa L.) is an essential leguminous forage with high nutrition and strong adaptability. The TIFY family is a plant-specific transcription factor identified in many plants. However, few reports have been reported on the phylogenetic analysis and gene expression profiling of TIFY family genes in alfalfa. Result: A total of 84 TIFY genes belonging to 4 categories were identified in alfalfa, including 58 MsJAZs, 18 MsZMLs, 4 MsTIFYs and 4 MsPPDs, respectively. qRT-PCR data from 8 genes in different tissues revealed that most MsTIFY genes were highly expressed in roots. The expression of MsTIFY14 was up-regulated after different times in both thrips-resistant and susceptible alfalfa after thrips feeding, and the expression of the remaining MsTIFYs had a strong correlation with the time of thrips feeding. Different abiotic stresses, including drought, salt, and cold, could induce or inhibit the expression of MsTIFY genes to varying degrees. In addition, the eight genes were all significantly up-regulated by JA and/or SA. Interestingly, MsTIFY77 was induced considerably by all the biotic, abiotic, or plant hormones (JA or SA) except ABA. Conclusion: Our study identified members of the TIFY gene family in alfalfa and analyzed their structures and possible functions. It laid the foundation for further research on the molecular functions of TIFYs in alfalfa. [ABSTRACT FROM AUTHOR]

Background: Phytochrome-interacting factors (PIFs) plays an important role in plants as hubs for intracellular signaling regulation. The PIF gene family has been identified and characterized in many plants, but alfalfa (Medicago sativa L.), an important perennial high-quality legume forage, has not been reported on the PIF gene family. Results: In this study, we presented the identification and characterization of five MsPIF genes in alfalfa (Medicago sativa L.). Phylogenetic analysis indicated that PIFs from alfalfa and other four plant species could be divided into three groups supported by similar motif analysis. The collinearity analysis of the MsPIF gene family showed the presence of two gene pairs, and the collinearity analysis with AtPIFs showed three gene pairs, indicating that the evolutionary process of this family is relatively conservative. Analysis of cis-acting elements in promoter regions of MsPIF genes indicated that various elements were related to light, abiotic stress, and plant hormone responsiveness. Gene expression analyses demonstrated that MsPIFs were primarily expressed in the leaves and were induced by various abiotic stresses. Conclusion: This study conducted genome-wide identification, evolution, synteny analysis, and expression analysis of the PIFs in alfalfa. Our study lays a foundation for the study of the biological functions of the PIF gene family and provides a useful reference for improving abiotic stress resistance in alfalfa. [ABSTRACT FROM AUTHOR]

Background: Alfalfa (Medicago sativa L.) is a globally important cool-season forage grass for semiarid regions and the demand for seeds is increasing with the increase of alfalfa planting area. However, its seed yield is often low or inconsistent. Methods: A field experiment was conducted to investigate the response of seed yield and yield components of alfalfa to different sowing dates and phosphorus (P) fertilization levels from 2020 to 2022. This research was conducted using a split-plot design in a randomized complete block, with four replications. The main plots were three sowing dates (20 July, 5 and 20 August) and subplots were five different rates (0, 40, 70, 100 and 130 kg P2O5 ha-1). Result: Our results revealed that delaying the sowing dates reduced seed yield, by influencing its yield components, except for the 1000-seed weight, but P fertilization enhanced the seed yield components affected by late sowing and increased seed yield, it is recommended to apply P fertilizer at 77.6~144.8 kg ha-1. Stem density and seeds per pod were the primary factors determining seed yield and they may be useful to consider in developing traits for high-seed-yield alfalfa. [ABSTRACT FROM AUTHOR]

Fusarium spp. can cause root rot in alfalfa, leading to the death of the whole plant, which seriously affects the yield and quality of alfalfa. This study used a Fusarium acuminatum strain labeled with green fluorescent protein (GFP) to observe the infection process of F. acuminatum on alfalfa by confocal fluorescence microscopy. The aim of this study was to reveal the infection mechanism of alfalfa Fusarium root rot at the cellular histological level. The results showed that conidia of F. acuminatum attached to the surface of the root and germinated at one day post-inoculation, the mycelium then entered the vascular bundle tissue of the alfalfa root at 5 days post-inoculation, reached the base of the plant stem at 14 days post-inoculation, and colonized the stem of the first and second compound leaf at 28 and 49 days post-inoculation, respectively. Moreover, the experiment, which sprayed a spore suspension, showed that the conidia of F. acuminatum could spread through the air to infect the pericarp and seed coat tissue of the pod. For the first time, we report the infection process of alfalfa Fusarium root rot caused by F. acuminatum and clarify that F. acuminatum can initially infect the root tissue of alfalfa, colonize the bottom stem of the plant through systematic infection, and eventually cause the plant to wilt and die. The results reveal the infection mechanism of F. acuminatum at the cell level via histology and provide theoretical support for the development of control strategies and key control technologies for alfalfa root rot. [ABSTRACT FROM AUTHOR]

To predict the role of the MsICE gene family in the response to abiotic stress, in this study, bioinformatics analysis and real-time fluorescence quantitative PCR were performed. Alfalfa (Medicago sativa) is one of the most economically valuable crops globally. Inducer of CBF expression (ICE), which is part of the basic helix–loop–helix (bHLH) transcription factor (TF) family, acts as a key regulator of cold tolerance. Despite this, there is little information available about ICE genes in alfalfa. Therefore, we studied the function of ICE TFs in alfalfa. We identified 11 MsICE genes from the alfalfa genome and classified them into two groups. Analysis of the protein motif and gene structure revealed relatively high conservation among subgroups of the tightly clustered MsICE genes. Through synteny analysis, we detected duplication events in the MsICE gene family, suggesting that the ICE gene family was formed through fragment duplications. All the MsICE proteins were located in the nucleus according to subcellular localization predictions. The promoter cis-regulatory elements of MsICE genes are largely involved in light (Box 4), hormone (ABRE), and stress (MYB) responses. The MsICE01/MsICE07/MsICE09/MsICE10/MsICE11 genes contained MYB- and MYC-binding motifs, indicating an association with abiotic stress. The specific expression patterns of MsICE genes in leaves were revealed by examining their expression patterns in different tissues. These findings suggest that these genes may sense external environmental changes through leaves. Abiotic stress can cause striking upregulation of MsICE07 (PCA score: −4.03) and MsICE10 (PCA score: −4.05) expression. In this study, candidate genes associated with cold stress were identified, and subsequent molecular biological analyses allowed elucidation of the biological functions of these genes in alfalfa. This research provides a theoretical foundation for enhancing alfalfa yield and quality under cold conditions. [ABSTRACT FROM AUTHOR]

In arid and semiarid regions, crop production has high irrigation water demands due to low precipitation. Efficient irrigation water management strategies can be developed using crop growth models to assess the effect of different irrigation management practices on crop productivity. The leaf area index (LAI) is an important growth parameter used in crop modeling. Measuring LAI requires specialized and expensive equipment not readily available for producers. Canopy cover (CC) and canopy height (CH) measurements, on the other hand, can be obtained with little effort using mobile devices and a ruler, respectively. The objective of this study was to determine the relationships between LAI, CC, and CH for fully and deficit-irrigated alfalfa (Medicago sativa L.). The LAI, CC, and CH measurements were obtained from an experiment conducted at the Valley Road Field Lab in Reno, Nevada, starting in the Fall of 2020. Three irrigation treatments were applied to two alfalfa varieties (Ladak II and Stratica): 100%, 80%, and 60% of full irrigation demands. Biweekly measurements of CC, CH, and LAI were collected during the growing seasons of 2021 and 2022. The dataset was randomly split into training and testing subsets. For the training subset, an exponential model and a simple linear regression (SLR) model were used to determine the individual relationship of CC and CH with LAI, respectively. Also, a multiple linear regression (MLR) model was implemented for the estimation of LAI with CC and CH as its predictors. The exponential model was fitted with a residual standard error (RSE) and coefficient of determination (R2) of 0.97 and 0.86, respectively. A lower performance was obtained for the SLR model (RSE = 1.03, R2 = 0.81). The MLR model (RSE = 0.82, R2 = 0.88) improved the performance achieved by the exponential and SLR models. The results of the testing indicated that the MLR performed better (RSE = 0.82, R2 = 0.88) than the exponential model (RSE = 0.97, R2 = 0.86) and the SLR model (RSE = 1.03, R2 = 0.82) in the estimation of LAI. The relationships obtained can be useful to estimate LAI when CC, CH, or both predictors are available and assist with the validation of data generated by crop growth models. [ABSTRACT FROM AUTHOR]

This study assessed the impact of phytoremediation on reducing the residual concentration of metolachlor in soil treated with doses of 530.7 and 1061.4 g/ha and its effect on microbial biodiversity in contaminated areas. For the plant species Avena sativa and Medicago sativa, a significant efficacy of 54.5 and 36.4% was observed in the dissipation of the herbicide, especially at higher doses. Although metolachlor application reduced soil microbial biodiversity, phytoremediating plants, especially M. sativa, promoted greater richness and distribution of microbial species, mitigating the negative effects of the herbicide. Principal component analysis revealed the influence of these plants and metolachlor on the composition of the microbial community. These results highlight the importance of phytoremediation in promoting soil biodiversity and reducing herbicide contamination, providing crucial insights for remediation strategies in contaminated areas. [ABSTRACT FROM AUTHOR]

Incorporating forage legumes into grasslands is a recommended climate change mitigation strategy, but accruing desired benefits from legumes is contingent upon their resilience when exposed to climate change factors (CCF). Our objective was to synthesize literature describing responses to CCF of a broad array of forage legume species, including annuals and perennials from both temperate and tropical/subtropical regions. Most‐represented species in the related literature include alfalfa (Medicago sativa L.), various Trifolium and Lotus species, rhizoma peanut (Arachis glabrata Benth.), and capitata stylo (Stylosanthes capitata Vogel). The data indicate that while CCF effects on forage legumes can be generalized, interactions among CCF, species, and site‐specific soil/climate conditions may cause variation from expected responses. In most instances, exposing forage legumes to CO2 enrichment (eCO2) increased forage accumulation (FA), but elevated temperature (eT) often reduced the magnitude of the positive response to eCO2. Photosynthetic acclimation to eCO2 occurs in non‐N‐fixing plants, but legume nodules are large C sinks and drive sustained increases in legume FA to eCO2. Legume N2 fixation and the proportion of legume N derived from N2 fixation increase with eCO2, but the magnitude of the increases lessens with eT. Legume nutritive value (NV) responses to eCO2 are less pronounced than FA responses, but decreased herbage N and greater nonstructural carbohydrate concentrations are common. Exposure to eT negatively affects NV unless intervals between defoliation events are shortened. Limited data on reproductive performance show CCF affect flower number, pollen grain morphology and viability, and behavior of pollinators, potentially influencing legume reproductive success. Core Ideas: Legume forage accumulation, N fixation, and proportion increase in mixed swards under enriched CO2.Elevated temperature decreases legume nutritive value and may ameliorate positive legume response to enriched CO2.Drought stress reduces legume forage accumulation, but drought effects on nutritive value are inconsistent.Climate change may affect legume flower number, nectar composition, pollinator visits, and reproductive success.Legume responses to climate change can be generalized, but interactions with species and local environments occur. [ABSTRACT FROM AUTHOR]

Crop height and biomass are the two important phenotyping traits to screen forage population types at local and regional scales. This study aims to compare the performances of multispectral and RGB sensors onboard drones for quantitative retrievals of forage crop height and biomass at very high resolution. We acquired the unmanned aerial vehicle (UAV) multispectral images (MSIs) at 1.67 cm spatial resolution and visible data (RGB) at 0.31 cm resolution and measured the forage height and above-ground biomass over the alfalfa (Medicago sativa L.) breeding trials in the Canadian Prairies. (1) For height estimation, the digital surface model (DSM) and digital terrain model (DTM) were extracted from MSI and RGB data, respectively. As the resolution of the DTM is five times less than that of the DSM, we applied an aggregation algorithm to the DSM to constrain the same spatial resolution between DSM and DTM. The difference between DSM and DTM was computed as the canopy height model (CHM), which was at 8.35 cm and 1.55 cm for MSI and RGB data, respectively. (2) For biomass estimation, the normalized difference vegetation index (NDVI) from MSI data and excess green (ExG) index from RGB data were analyzed and regressed in terms of ground measurements, leading to empirical models. The results indicate better performance of MSI for above-ground biomass (AGB) retrievals at 1.67 cm resolution and better performance of RGB data for canopy height retrievals at 1.55 cm. Although the retrieved height was well correlated with the ground measurements, a significant underestimation was observed. Thus, we developed a bias correction function to match the retrieval with the ground measurements. This study provides insight into the optimal selection of sensor for specific targeted vegetation growth traits in a forage crop. [ABSTRACT FROM AUTHOR]

This work aimed to study the effect of adding alfalfa (Medicago sativa L.) and Atriplex halimus L. on biochemical properties, microbial flora, ferment silage and solar-dried cactus fruit scraps. Alfalfa and Atriplex were added to cactus silage to improve nutrient quality. Three mixtures were prepared: M1 based on cactus, M2 based on cactus and alfalfa, and M3 based on cactus and Atriplex. Bran and wheat straw were added to the mixtures to improve the moisture content. After adding alfalfa and Atriplex, the protein content of the pre-ensiling material increased (p < 0.05), reaching 10.63% DM for M1, 16.35% DM for M2, and 17.22% DM for M3. During ensilage, a significant amount of sugar and protein was degraded. All silage treatments achieved stable pH values (M1: 4.35, M2: 4.55, and M3: 4.54). The M2 and M3 treatments had the highest amount of protein. After drying, the protein content of the dried treatments was higher than that of the ensiled treatments (8.05% DM for M1, 14.96% DM for M2, and 15.36% DM for M3). In both storage methods, the microbial population declined. However, silage reduces coliforms, yeasts and moulds more efficiently than solar drying. The drying operation preserves remarkedly the nutritional quality of the mixtures while ensiling reduces the number of undesirable microorganisms. [ABSTRACT FROM AUTHOR]

Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Medicago truncatula is a key model plant for studying legume plants, particularly alfalfa (Medicago sativa), due to its well-defined genetic background. Plant-specific GASA (Gibberellic Acid Stimulated Arabidopsis) genes play various roles in plant growth and development, abiotic stress, and hormone responses. However, limited information is available on GASA research in Medicago. In this study, 26 MtGASAs were identified and analyzed for its structure, evolution, and expressions. Sequence alignments and phylogeny revealed that 26 MtGASAs containing conserved GASA domains were classified into three clades. The chromosomal locations and gene synteny revealed segmental and tandem repetition evolution. Analysis of cis-regulatory elements indicates that family members likely influence various hormone signaling pathways and stress-related mechanisms. Moreover, the RNA-seq and qRT-PCR analyses revealed that 26 MtGASAs were extensively involved in abiotic stresses and hormone responses. Notably, seven MtGASA genes (MtGASA1, 10, 12, 17, 23, 25 and 26) were all dramatically activated by NaCl and Mannitol treatments, and four MtGASAs (MtGASA7, 10, 23 and 24) were significant activated by GA3, PBZ, ABA, and MeJA treatments. Collectively, this study is the first to identify and describe GASA genes in Medicago on a genome-wide scale. The results establish a basis for functional characterization, showing that these proteins are essential in responding to various abiotic stresses and hormonal signals. [ABSTRACT FROM AUTHOR]

Due to the increasing demand for green fodder in Iraq to sustain livestock, alfalfa cultivation primarily focuses on fodder production, while seed yield is deemed of secondary importance. Meeting this demand requires implementing diverse agricultural strategies aimed at enhancing alfalfa seed yield, notably through the application of plant growth regulators (growth retardants). Therefore, this study aims to know the possibility of increasing seed yield and determining the appropriate spraying stage and concentration using the growth inhibitor Ethephon to increase seed yield of the local variety of alfalfa. A field experiment was carried out from October 2021 to the end of August 2022 at the Experimental Field at the College of Agricultural Engineering Sciences, University of Baghdad, Iraq. The main objective of the study was to find out the effect of Ethephon concentrations (0, 250, 500 and 750) mg/L and three stages of foliar application (vegetative growth, beginning appearance of flower buds and 50% flowering) on seed yield and its components of alfalfa local variety. The layout of the experiment was Randomized Complete Block Design (RCBD) arranged in a split plot, with three replications. Foliar application stages were used as main plots while the Ethephon concentrations were used as sub-plots. Spraying of Ethephon at 750 mg/L resulted in a significant increment in the number of stems m-2, number of ripe racemes per stem, number of pods per raceme, biological yield, seed yield and harvest index. The highest seed yield (550.3 kg/ha) of alfalfa was obtained when plants were sprayed with Ethephon 750 mg/L at vegetative growth, which produces significantly highest number of racemes per stem and number of pods per raceme compared to other concentrations of Ethephon. The mean seed yield amounted to 453.2 kg/ha and it was significantly higher compared to Ethephon concentrations of 0 and 250 mg/L but at par with 750 mg/L concentration of Ethephon. [ABSTRACT FROM AUTHOR]

The influence of molasses (M) and Lactobacillus rhamnosus (LR) on fermentation products, microbial communities, and metabolites in big-bale alfalfa silage was investigated. Alfalfa (Medicago sativa L.) was harvested at the third growth stage during the flowering stage in the experimental field of Linhui Grass Company from Tongliao City, Inner Mongolia. An alfalfa sample without additives was used as a control (C). M (20 g/kg) and LR (106 cfu/g) were added either alone or in combination. Alfalfa was fermented for 7, 14, and 56 d. Lactic acid content in the M, LR, and MLR groups increased, whereas the pH value and butyric acid, 2,3-butanediol, and ethanol contents decreased compared to those of C group after 7, 14, and 56 d of fermentation. A two-way analysis of variance (ANOVA) was performed to estimate the results. The LR group exhibited increased Lactobacillus abundance, whereas the M and MLR groups showed increased Weissella abundance compared to the C group. The relative contents of amino acids (tyrosine, isoleucine, threonine, arginine, valine, and citrulline) in the M and MLR groups were higher than those in the C group. During fermentation, the M, LR, and MLR groups showed decreased phenylalanine, isoleucine, and ferulic acid contents. Amino acids such as isoleucine and L-aspartic acid were positively correlated with Lactobacillus but negatively correlated with Weissella. In conclusion, combining high-throughput sequencing and liquid chromatography–mass spectrometry during anaerobic alfalfa fermentation can reveal new microbial community compositions and metabolite profiles, supporting the application of M, LR, and MLR as feed fermentation agents. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) is an economically important forage legume. As a result of the extensive and intensive cultivation of alfalfa planting and the gradual expansion of planting areas, pest occurrence has increased in frequency. Aphids are one of the main pests that threaten the growth and productivity of alfalfa. After an aphid outbreak, alfalfa yield and quality are commonly greatly reduced. At present, there are few studies on alfalfa plants infested with aphids, so it is imperative to study the regulatory mechanisms of aphid infestation tolerance in alfalfa. In this study, alfalfa plants from the variety "Wudi" were investigated, and pea aphids were selected for inoculation. The transcriptome and metabolome data were analyzed at three time points (0 d, 1 d, and 4 d), revealing 3458 differentially expressed genes and 358 differential metabolites. Trend analysis of DEGs and DAMs revealed that the former were significantly enriched in three distinct trends, whereas the latter were significantly enriched in only one. Moreover, 117 important hub genes associated with alfalfa response to aphid infestation were screened by WGCNA. By mapping DEGs and DAMs to KEGG pathways, it was found that the "phenylpropanoid biosynthesis", "flavonoid biosynthesis", and "isoflavonoid biosynthesis" pathways play an important role in alfalfa responses to aphid infestation. These results further elucidate the regulatory mechanism of alfalfa in response to aphid infestation and provide valuable information for breeding new aphid-resistant plant varieties. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Education & Science is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract The adaptability of insects to hosts has long been a focal point in the study of insect-plant interactions. The pea aphid (Acythosiphon pisum), a significant pest of numerous leguminous crops, not only inflicts direct economic losses but also disseminates various plant viruses. To understand how pea aphids adapt to diverse alfalfa varieties. We analyzed the differentially expressed genes (DEGs) of pea aphids in distinct alfalfa varieties using transcriptome sequencing, and subsequently conducted functional validation of these genes. Comparative analysis between pea aphids feeding on susceptible and resistant strains revealed that DEGs in aphids feeding on resistant strains were primarily associated with transcriptional enrichment in the sugar, amino acid, protein, and lipid metabolism pathways. Fourteen DEGs related to adaptation of the pea aphid to alfalfa were chosen, including five carboxylesterases (CarE), four cytochrome P450s, three glutathione S-transferases, and two peroxidases (POD). RT-qPCR results indicated significant up-regulation of two carboxylesterase genes and two peroxidase genes after 24 h of feeding resistant alfalfa (Gannong 5, GN5) compared to the susceptible varieties (Hunter River, LRH), particularly highlighting the high expression levels of ApCarE4 and ApPOD3. Simultaneously, RNAi-induced knockdown of ApCarE4 and ApPOD3 led to a higher mortality of pea aphids in the alfalfa Hunter River. These results indicate that ApPOD3 and ApCarE4 are involved in the detoxification of metabolic functions in the adaptation of pea aphids to host switching. These findings contribute to the understanding of pea aphid adaptation to host plants and lay a foundation for further exploration of the physiological roles of carboxylesterase and peroxidase genes in pea aphids.

Abstract Background Drought stress is a major limiting factor that affects forage yields, and understanding the drought resistance mechanism of plants is crucial for improving crop yields in arid areas. Alfalfa (Medicago sativa L.) is the most important legume plant, mainly planted in arid and semi-arid areas. However, the adaptability of alfalfa to drought stress and its physiological and molecular mechanisms of drought resistance remains unclear. Results In this study, we analyzed the physiological and transcriptome responses of alfalfa cultivars with different drought resistances (drought-sensitive Gannong No. 3 (G3), drought-resistant Gannong No. 8 (G8), and strong drought-resistant Longdong (LD)) under drought stress at 0, 6, 12, and 24 h. LD had higher catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities and a higher soluble protein content, lower malondialdehyde (MDA) content, a lower O2 ·− production rate, and a lower H2O2 content than G8 and G3 (P

Abstract MicroRNA (miRNA) is a type of endogenous non-coding small RNA, which is abundant in living organisms. miRNAs play an important role in regulating gene expression and myriad cellular processes by binding to target messenger RNAs through complementary base pairing, and cross-species regulation mammalian cells by plant-derived xeno-miRNAs has been described. Here, we examined the miRNA species in two alfalfa (Medicago sativa, lucerne) cultivars commonly grown in Ningxia, China: cv. Zhongmu 1 and cv. Xinyan 52. Both cultivars have good salt and drought resistance. We found that the miRNA profiles were similar between the cultivars, with a slightly higher number of miRNAs present in the newer cv. Xinyan 52, which may contribute to its improved salt and drought tolerance. miRNAs were stable during drying, and some miRNAs were increased in dry versus fresh alfalfa, suggesting some miRNAs may be upregulated during drying. Alfalfa-derived miRNAs could be detected in exosomes from serum and whey collected from dairy cows, confirming the ability of the exogenous miRNAs (xeno-miRNAs) to enter the circulation and reach the mammary epithelium. In vitro studies confirmed that overexpression of mtr-miR156a could downregulate expression of Phosphatase 2 Regulatory Subunit B'gamma ( PPP2R5D) and Phosphoinositide-3-kinase Regulatory Subunit 2 (PIK3R2). Overexpression of mtr-miR156a also modulated PI3K–AKT–mTOR signaling as well as the casein content of milk produced by bovine mammary epithelial cells. Based on the known roles of PPP2R5D and PIK3R2 in regulating the PI3K–AKT–mTOR pathway as well as the effect of PI3K–AKT–mTOR on milk protein content, our findings implicate alfalfa-derived miR156a as a new cross-species regulator of milk quality in dairy cows.

Abstract Background Alfalfa (Medicago sativa L.) is an essential leguminous forage with high nutrition and strong adaptability. The TIFY family is a plant-specific transcription factor identified in many plants. However, few reports have been reported on the phylogenetic analysis and gene expression profiling of TIFY family genes in alfalfa. Result A total of 84 TIFY genes belonging to 4 categories were identified in alfalfa, including 58 MsJAZs, 18 MsZMLs, 4 MsTIFYs and 4 MsPPDs, respectively. qRT-PCR data from 8 genes in different tissues revealed that most MsTIFY genes were highly expressed in roots. The expression of MsTIFY14 was up-regulated after different times in both thrips-resistant and susceptible alfalfa after thrips feeding, and the expression of the remaining MsTIFYs had a strong correlation with the time of thrips feeding. Different abiotic stresses, including drought, salt, and cold, could induce or inhibit the expression of MsTIFY genes to varying degrees. In addition, the eight genes were all significantly up-regulated by JA and/or SA. Interestingly, MsTIFY77 was induced considerably by all the biotic, abiotic, or plant hormones (JA or SA) except ABA. Conclusion Our study identified members of the TIFY gene family in alfalfa and analyzed their structures and possible functions. It laid the foundation for further research on the molecular functions of TIFYs in alfalfa.

Abstract Background Drought stress restricts the growth, distribution and productivity of alfalfa (Medicago sativa L.). In order to study the response differences of alfalfa cultivars to drought stress, we previously carried out physiological and molecular comparative analysis on two alfalfa varieties with contrasting drought resistance (relatively drought-tolerant Longdong and drought-sensitive Algonquin). However, the differences in proteomic factors of the two varieties in response to drought stress still need to be further studied. Therefore, TMT-based quantitative proteomic analysis was performed using leaf tissues of the two alfalfa cultivars to identify and uncover differentially abundant proteins (DAPs). Results In total, 677 DAPs were identified in Algonquin and 277 in Longdong under drought stress. Subsequently, we conducted various bioinformatics analysis on these DAPs, including subcellular location, functional classification and biological pathway enrichment. The first two main COG functional categories of DAPs in both alfalfa varieties after drought stress were ‘Translation, ribosomal structure and biogenesis’ and ‘Posttranslational modification, protein turnover, chaperones’. According to KEGG database, the DAPs of the two alfalfa cultivars after drought treatment were differentially enriched in different biological pathways. The DAPs from Algonquin were enriched in ‘photosynthesis’ and ‘ribosome’. The pathways of ‘linoleic acid metabolism’, ‘protein processing in endoplasmic reticulum’ and ‘RNA transport’ in Longdong were significantly enriched. Finally, we found significant differences in DAP enrichment and expression patterns between Longdong and Algonquin in glycolysis/glycogenesis, TCA cycle, photosynthesis, protein biosynthesis, flavonoid and isoflavonoid biosynthesis, and plant-pathogen interaction pathway after drought treatment. Conclusions The differences of DAPs involved in various metabolic pathways may explain the differences in the resistance of the two varieties to drought stress. These DAPs can be used as candidate proteins for molecular breeding of alfalfa to cultivate new germplasm with more drought tolerance to adapt to unfavorable environments.

Abstract High temperature stress is one of the most severe forms of abiotic stress in alfalfa. With the intensification of climate change, the frequency of high temperature stress will further increase in the future, which will bring challenges to the growth and development of alfalfa. Therefore, untargeted metabolomic and RNA-Seq profiling were implemented to unravel the possible alteration in alfalfa seedlings subjected to different temperature stress (25 ℃, 30 ℃, 35 ℃, 40 ℃) in this study. Results revealed that High temperature stress significantly altered some pivotal transcripts and metabolites. The number of differentially expressed genes (DEGs) markedly up and down-regulated was 1876 and 1524 in T30_vs_CK, 2, 815 and 2667 in T35_vs_CK, and 2115 and 2, 226 in T40_vs_CK, respectively. The number for significantly up-regulated and down-regulated differential metabolites was 173 and 73 in T30_vs_CK, 188 and 57 in T35_vs_CK, and 220 and 66 in T40_vs_CK, respectively. It is worth noting that metabolomics and transcriptomics co-analysis characterized enriched in plant hormone signal transduction (ko04705), glyoxylate and dicarboxylate metabolism (ko00630), from which some differentially expressed genes and differential metabolites participated. In particular, the content of hormone changed significantly under T40 stress, suggesting that maintaining normal hormone synthesis and metabolism may be an important way to improve the HTS tolerance of alfalfa. The qRT-PCR further showed that the expression pattern was similar to the expression abundance in the transcriptome. This study provides a practical and in-depth perspective from transcriptomics and metabolomics in investigating the effects conferred by temperature on plant growth and development, which provided the theoretical basis for breeding heat-resistant alfalfa.

Abstract Background We aimed to gain insight into the response mechanism of alfalfa (Medicago sativa) to drought stress by recognizing and analyzing drought-responsive genes in the roots of different root types of alfalfa. The rhizomatous-rooted M. sativa cv.‘Qingshui’ (QS), tap-rooted M. sativa cv.‘Longdong’ (LD), and creeping-rooted M. varia cv. ‘Gongnong No. 4’ (GN) were used to analyze the transcriptome information and physiological characteristics of the root systems of the cultivars under simulated drought stress using PEG-6000. Results It was found that aridity caused a significant increase in the content of osmotic stress substances and antioxidant enzyme activity. The content of malondialdehyde (MDA) in QS was lower than that in LD and GN under moisture stress, indicating a stronger accumulation capacity of osmotic regulatory substances. Based on sequencing results, 14,475, 9336, and 9243 upregulated DEGs from QS, LD, and GN were annotated into 26, 29, and 28 transcription factor families, respectively. QS showed more DEGs than LD and GN. KEGG enrichment analysis identified that DEGs were significantly enriched in metabolic pathways such as amino acid biosynthesis, phenylpropanoid biosynthesis, plant hormone signaling transduction, and MAPK pathways. This suggests a strong correlation between these pathways and drought stress. The results also show that genes associated with ABA hormone signaling (MS. gene93372, MS. gene072046, and MS. gene012975) are crucial for plant’s adaptation to drought stress. Conclusions These genes, such as serine/threonine protein kinases and abscisic acid receptors, play a crucial role in plant hormone signaling and MAPK pathways. They could serve as potential candidate genes for drought resistance research in alfalfa, providing a molecular foundation for studying drought resistance. Graphical Abstract

Abstract Background Daye No.3 is a novel cultivar of alfalfa (Medicago sativa L.) that is well suited for cultivation in high-altitude regions such as the Qinghai‒Tibet Plateau owing to its high yield and notable cold resistance. However, the limited availability of transcriptomic information has hindered our investigation into the potential mechanisms of cold tolerance in this cultivar. Consequently, we conducted de novo transcriptome assembly to overcome this limitation. Subsequently, we compared the patterns of gene expression in Daye No. 3 during cold acclimatization and exposure to cold stress at various time points. Results A total of 15 alfalfa samples were included in the transcriptome assembly, resulting in 141.97 Gb of clean bases. A total of 441 DEGs were induced by cold acclimation, while 4525, 5016, and 8056 DEGs were identified at 12 h, 24 h, and 36 h after prolonged cold stress at 4 °C, respectively. The consistency between the RT‒qPCR and transcriptome data confirmed the accuracy and reliability of the transcriptomic data. KEGG enrichment analysis revealed that many genes related to photosynthesis were enriched under cold stress. STEM analysis demonstrated that genes involved in nitrogen metabolism and the TCA cycle were consistently upregulated under cold stress, while genes associated with photosynthesis, particularly antenna protein genes, were downregulated. PPI network analysis revealed that ubiquitination-related ribosomal proteins act as hub genes in response to cold stress. Additionally, the plant hormone signaling pathway was activated under cold stress, suggesting its vital role in the cold stress response of alfalfa. Conclusions Ubiquitination-related ribosomal proteins induced by cold acclimation play a crucial role in early cold signal transduction. As hub genes, these ubiquitination-related ribosomal proteins regulate a multitude of downstream genes in response to cold stress. The upregulation of genes related to nitrogen metabolism and the TCA cycle and the activation of the plant hormone signaling pathway contribute to the enhanced cold tolerance of alfalfa.

The current paper has presented the study conducted in the alfalfa collection nursery on the experimental plots of the Agricultural Research Center “Donskoy” in the period from 2021 to 2023. The purpose of the study was to determine the dependence of green mass productivity of collection alfalfa samples on the morphological traits of plants. The object of the study was a collection of alfalfa, consisting of 105 samples developed created at FSBSI ARC “Donskoy”. Sowing was carried out manually, with row spacing of 20 cm. The area of the plots was 1 m2 , with double repetition. The seed-sowing rate for alfalfa was 2 g/m2 . The standard variety was ‘Rostovskaya 90’. As a result, there was revealed that over 3 years the highest mean productivity of alfalfa green mass was formed in two groups of samples according to plant height: 1) 80–85 cm – 8.10 kg/m2 (the samples ‘SGP-424’, ‘SPCh 401/2000’); 2) 105–110 cm – 7.88 kg/m2 (‘SGP-162’, ‘SGP-189’, ‘SGP-175’, etc.). This indicator of alfalfa productivity was the highest (7.81 kg/m2 ) in the optimal values of tillering (15–20 pcs.). The highest alfalfa green mass productivity was formed by the samples with foliage of 48–50 % and 42–44 %. The green mass productivity of alfalfa samples varied within the range of 3.33–10.49 kg/m2 . There have been identified 7 samples which productivity ranged from 9.09 kg/m2 to 10.49 kg/m2. There have been selected the samples that combined the highest green mass productivity with tallness and high foliage of plants, as well as the greatest leaf length and hay yields.