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Leaves are the most significant parts of forage crops such as alfalfa. Senescence is the terminal stage of leaf development and is controlled by an integrated myriad of endogenous signals and environmental stimuli. WRKY transcription factors (TFs) play essential roles in regulating leaf senescence; however, only a few studies on the analysis and identification of the WRKY TF family in Medicago Sativa have been reported. In this study, we identified 198 WRKY family members from the alfalfa (M. sativa L.) cultivar 'XinjiangDaye' using phylogenetic analysis and categorized them into three subfamilies, Groups I, II, and III, based on their structural characteristics. Group II members were further divided into five subclasses. In addition, several hormone- and stress-related cis-acting elements were identified in the promoter regions of MsWRKYs. Furthermore, 14 aging-related MsWRKYs genes from a previous transcriptome in our laboratory were selected for RT-qPCR validation of their expression patterns, and subsequently cloned for overexpression examination. Finally, MsWRKY5, MsWRKY66, MsWRKY92, and MsWRKY141 were confirmed to cause leaf yellowing in Nicotiana benthaminana using a transient expression system. Our findings lay a groundwork for further studies on the mechanism of M. sativa leaf aging and for the creation of new germplasm resources. [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]

Drought may impact plant–soil biotic interactions in ways that modify aboveground herbivore performance, but the outcomes of such biotic interactions under future climate are not yet clear. We performed a growth chamber experiment to assess how long‐term, drought‐driven changes in belowground communities influence plant growth and herbivore performance using a plant–soil feedback experimental framework. We focussed on two common pasture legumes—lucerne, Medicago sativa L., and white clover, Trifolium repens L. (both Fabaceae)—and foliar herbivores—cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), and two‐spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). Soil was collected from a field facility where rainfall had been manipulated for 6 years, focussing on treatments representing ambient rainfall and prolonged drought (50% reduction relative to ambient), to consider the effects of biological legacies mediated by the prolonged drought. All soils were sterilized and re‐inoculated to establish the respective home (i.e. where a given plant is cultivated in its own soil) and away (i.e. where a given plant is cultivated in another species' soil) treatments in addition to a sterile control. We found that the relative growth rate (RGR) and relative consumption of larvae were significantly lower on lucerne grown in soil with ambient rainfall legacies conditioned by white clover. Conversely, the RGR of insect larvae was lower on white clover grown in soil with prolonged drought legacies conditioned by lucerne. Two‐spotted spider mite populations and area damage (mm2) were significantly reduced on white clover grown in lucerne‐conditioned soil in drought legacies. The higher number of nodules found on white clover in lucerne‐conditioned soil suggests that root–rhizobia associations may have reduced foliar herbivore performance. Our study provides evidence that foliar herbivores are affected by plant–soil biotic interactions and that prolonged drought may influence aboveground–belowground linkages with potential broader ecosystem impacts. [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]

Background: Alfalfa (Medicago sativa L.) is the most widely planted legume forage and one of the most economically valuable crops in the world. Serine hydroxymethyltransferase (SHMT), a pyridoxal phosphate-dependent enzyme, plays crucial roles in plant growth, development, and stress responses. To date, there has been no comprehensive bioinformatics investigation conducted on the SHMT genes in M. sativa. Results: Here, we systematically analyzed the phylogenetic relationship, expansion pattern, gene structure, cis-acting elements, and expression profile of the MsSHMT family genes. The result showed that a total of 15 SHMT members were identified from the M. sativa genome database. Phylogenetic analysis demonstrated that the MsSHMTs can be divided into 4 subgroups and conserved with other plant homologues. Gene structure analysis found that the exons of MsSHMTs ranges from 3 to 15. Analysis of cis-acting elements found that each of the MsSHMT genes contained different kinds of hormones and stress-related cis-acting elements in their promoter regions. Expression and function analysis revealed that MsSHMTs expressed in all plant tissues. qRT-PCR analysis showed that MsSHMTs induced by ABA, Salt, and drought stresses. Conclusions: These results provided definite evidence that MsSHMTs might involve in growth, development and adversity responses in M. sativa, which laid a foundation for future functional studies of MsSHMTs. [ABSTRACT FROM AUTHOR]

Biostimulants such as ascorbic acid, known as vitamin C, have been reported to have numerous positive roles in plant tolerance to abiotic stresses. However, little is known about the biostimulant effects of ascorbic acid on alfalfa (Medicago sativa). Accordingly, a pot experiment was conducted to investigate the effects of 1 mM ascorbic acid, applied as foliar spray, on the salt tolerance of a Moroccan alfalfa population Demnate 201. One month-old M. sativa seedlings were exposed to 200 mM NaCl for four weeks with or without 1 mM of exogenous ascorbic acid treatment. The results showed that salinity stress significantly (p < 0.001) reduced plant biomass, disturbed photosynthesis-related parameters and induced oxidative stress. However, ascorbic acid foliar spray counteracted the observed negative effects of salinity. It significantly (p < 0.001) improved plant growth and photosynthetic parameters. Besides, stress indicators, including Na+ in shoot and root, hydrogen peroxide and electrolyte leakage, were significantly reduced by 42%, 29%, 12% and 34%, respectively, in treated and salt-stressed alfalfa plants. Interestingly, the decrease in oxidative stress markers was positively correlated to the ability of ascorbic acid to induce the accumulation of flavonoids and to increase the antioxidant activity of guaiacol peroxidase. Furthermore, compatible solutes, such as proline and soluble sugars, were found higher especially in salt-stressed alfalfa plants treated with 1 mM ascorbic acid. Our findings showed that ascorbic acid supply could be an eco-friendly and sustainable technique to mitigate the toxic effect of salt and could improve alfalfa forage production when grown in salt-affected soils. [ABSTRACT FROM AUTHOR]

Modern agriculture is facing a number of challenges, among which the efficient use of fertilisers to provide plants with the necessary nutrients is of particular importance. In particular, the growing focus on sustainable agriculture and the need to preserve soil resources underline the relevance of research into optimising fertiliser use. The purpose of the study was to identify effective methods of fertiliser use to provide plants with nutrients. To achieve this goal, the effectiveness of green manure, organic (manure) and mineral fertilisers under Sorghum and perennial grasses (Medicago sativa, Elytrigia, Agropyron) on saline soil was investigated. The study founded that organic fertilisers, such as manure and green manure, contribute to a more even supply of nutrients to plants compared to mineral fertilisers. This is particularly important in the context of nitrogen and phosphorus balance, where Medicago sativa has a positive nitrogen balance due to its ability to symbiotically fix nitrogen. Other crops studied, such as Elytrigia, Agropyron and Sorghum, showed a negative nitrogen balance on the control variant, but the application of N100P50K110 mineral fertiliser resulted in a positive nitrogen balance. The replacement of mineral fertilisers with organic fertilisers such as manure and green manure increased nitrogen inputs but also increased nitrogen removal from the soil due to the increase in phytomass yields. The results of the study confirmed the importance of transitioning to a more sustainable and environmentally friendly way of production in agriculture, which will ensure the development of the industry and increase its competitiveness in the international market. The practical significance of the study is to provide farmers with the opportunity to increase the yield of their crops and at the same time preserve soil fertility through the optimal use of nutrients contained in fertilisers. [ABSTRACT FROM AUTHOR]

Morocco faces a growing waste issue due to population growth, economic expansion, and industrialization, leading to environmental concerns, especially regarding leachate. From 1986 to 2022, Casablanca's main landfill produced a total of 800,000 m² of leachate, which was stored in evaporation ponds, posing significant environmental risks. Our research not only concentrates on traditional chemical analyses for leachate assessments but also emphasizes ecosystem interactions. Phytotoxicity tests assess the impact of contaminants, particularly heavy metals, complementing traditional chemical analyses. Our study investigated the accumulation of these contaminants in the soil and their subsequent transfer to plant tissues. This research aimed to examine the accumulation of heavy metals, including Pb, Cd, and Hg, in soils irrigated with varying leachate concentrations (C0: control; C1: 5%; C2: 7%; C3: 10%; and C4: 15%). The experiments involved the cultivation of M. sativa plants under open-field conditions. To assess the transfer of metals from soil to plant tissues, the transfer factor (TF) index was calculated. Our findings revealed that Pb, Cd, and Hg exhibited transfer factor ranges of 0.55-0.93, 0.07 to 0.21, and 0.1 to 0.37, respectively. The accumulation of heavy metals at the different leachate concentrations followed the order Pb > Hg > Cd. [ABSTRACT FROM AUTHOR]

This study investigated using Hedychium gardnerianum, an invasive plant, combined with dehydrated alfalfa (Medicago sativa) with varying percentages of alfalfa: control (0% alfalfa), T1 (10%), T2 (20%), T3 (30%), and T4 (40%), to enhance nutritional parameters in silage for ruminants in the Azores. Samples were vacuum-sealed and stored for 45 days at room temperature to promote anaerobic fermentation. Chemical analysis showed that alfalfa addition increased dry matter (DM) from 12.33% to 48.04% and crude protein from 11.34% DM to 24.63% DM. Insoluble fiber levels decreased, enhancing digestibility. In vitro fermentation indicated higher gas production in 40% alfalfa silage, suggesting faster fermentation. In conclusion, incorporating dehydrated alfalfa into Hedychium gardnerianum silage particularly enhances nutritional quality and digestibility. This approach offers a practical solution for ruminant feed in the Azores, particularly during feed scarcity periods. [ABSTRACT FROM AUTHOR]

Abstract 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 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.

Modern agriculture is facing a number of challenges, among which the efficient use of fertilisers to provide plants with the necessary nutrients is of particular importance. In particular, the growing focus on sustainable agriculture and the need to preserve soil resources underline the relevance of research into optimising fertiliser use. The purpose of the study was to identify effective methods of fertiliser use to provide plants with nutrients. To achieve this goal, the effectiveness of green manure, organic (manure) and mineral fertilisers under Sorghum and perennial grasses (Medicago sativa, Elytrigia, Agropyron) on saline soil was investigated. The study founded that organic fertilisers, such as manure and green manure, contribute to a more even supply of nutrients to plants compared to mineral fertilisers. This is particularly important in the context of nitrogen and phosphorus balance, where Medicago sativa has a positive nitrogen balance due to its ability to symbiotically fix nitrogen. Other crops studied, such as Elytrigia, Agropyron and Sorghum, showed a negative nitrogen balance on the control variant, but the application of N100P50K110 mineral fertiliser resulted in a positive nitrogen balance. The replacement of mineral fertilisers with organic fertilisers such as manure and green manure increased nitrogen inputs but also increased nitrogen removal from the soil due to the increase in phytomass yields. The results of the study confirmed the importance of transitioning to a more sustainable and environmentally friendly way of production in agriculture, which will ensure the development of the industry and increase its competitiveness in the international market. The practical significance of the study is to provide farmers with the opportunity to increase the yield of their crops and at the same time preserve soil fertility through the optimal use of nutrients contained in fertilisers

Abstract Background Alfalfa (Medicago sativa L.) is the most widely planted legume forage and one of the most economically valuable crops in the world. Serine hydroxymethyltransferase (SHMT), a pyridoxal phosphate-dependent enzyme, plays crucial roles in plant growth, development, and stress responses. To date, there has been no comprehensive bioinformatics investigation conducted on the SHMT genes in M. sativa. Results Here, we systematically analyzed the phylogenetic relationship, expansion pattern, gene structure, cis-acting elements, and expression profile of the MsSHMT family genes. The result showed that a total of 15 SHMT members were identified from the M. sativa genome database. Phylogenetic analysis demonstrated that the MsSHMTs can be divided into 4 subgroups and conserved with other plant homologues. Gene structure analysis found that the exons of MsSHMTs ranges from 3 to 15. Analysis of cis-acting elements found that each of the MsSHMT genes contained different kinds of hormones and stress-related cis-acting elements in their promoter regions. Expression and function analysis revealed that MsSHMTs expressed in all plant tissues. qRT-PCR analysis showed that MsSHMTs induced by ABA, Salt, and drought stresses. Conclusions These results provided definite evidence that MsSHMTs might involve in growth, development and adversity responses in M. sativa, which laid a foundation for future functional studies of MsSHMTs.

Morocco faces a growing waste issue due to population growth, economic expansion, and industrialization, leading to environmental concerns, especially regarding leachate. Casablanca’s main landfill, which is operational from 1986 to 2022, produces more than 800,000 m3/year of leachate, posing significant environmental risks. Our research not only concentrates on traditional chemical analyses for leachate assessments but also emphasizes ecosystem interactions. Phytotoxicity tests assess the impact of contaminants, particularly heavy metals, complementing traditional chemical analyses. Our study investigated the accumulation of these contaminants in the soil and their subsequent transfer to plant tissues. This research aimed to examine the accumulation of heavy metals, including Pb, Cd, and Hg, in soils irrigated with varying leachate concentrations (C0: control; C1: 5%; C2: 7%; C3: 10%; and C4: 15%). The experiments involved the cultivation of M. sativa plants under open-field conditions. To assess the transfer of metals from soil to plant tissues, the transfer factor (TF) index was calculated. Our findings revealed that Pb, Cd, and Hg exhibited transfer factor ranges of 0.55-0.93, 0.07 to 0.21, and 0.1 to 0.37, respectively. The accumulation of heavy metals at the different leachate concentrations followed the order Pb > Hg > Cd.

Abstract Alfalfa is widely recognized as an important forage crop. To understand the morphological characteristics and genetic basis of seed morphology in alfalfa, we screened 318 Medicago spp., including 244 Medicago sativa subsp. sativa (alfalfa) and 23 other Medicago spp., for seed area size, length, width, length-to-width ratio, perimeter, circularity, the distance between the intersection of length & width (IS) and center of gravity (CG), and seed darkness & red–green–blue (RGB) intensities. The results revealed phenotypic diversity and correlations among the tested accessions. Based on the phenotypic data of M. sativa subsp. sativa, a genome-wide association study (GWAS) was conducted using single nucleotide polymorphisms (SNPs) called against the Medicago truncatula genome. Genes in proximity to associated markers were detected, including CPR1, MON1, a PPR protein, and Wun1(threshold of 1E−04). Machine learning models were utilized to validate GWAS, and identify additional marker-trait associations for potentially complex traits. Marker S7_33375673, upstream of Wun1, was the most important predictor variable for red color intensity and highly important for brightness. Fifty-two markers were identified in coding regions. Along with strong correlations observed between seed morphology traits, these genes will facilitate the process of understanding the genetic basis of seed morphology in Medicago spp.

This study aims to compare the physiological and biochemical behaviour of four varieties of Medicago sativa differing in origin: Tamacine (Algerian), Triade (Italian), Maxigrowth (American), and Zanandrea (Australian). These varieties were subjected to saline stress (100 mM L-1) for 45 days at the early flowering stage. The results show that salinity had a depressive effect on all four varieties, reducing both fresh and dry matter. A decrease of more than 15% in dry weight was observed in the Tamacine genotype, but this reduction was more pronounced in the Zanandrea genotype. A significant increase in proline and soluble sugars content was recorded in the leaves (p = 0.000) under saline stress. Tamacine appears to be the most resistant variety, demonstrating biochemical adaptation by accumulating more osmoprotectants. [ABSTRACT FROM AUTHOR]

As the number of livestock increased in the years following English colonisation of Australia in 1788, the need for nutritious fodder, including lucerne (Medicago sativa), grew. One of the first diseases found on lucerne was a leaf spot which was collected in 1879 by George Bancroft, a physician and naturalist, in a suburb of Brisbane. The Queensland Government Botanist Frederick Manson Bailey sent a specimen to the prominent English mycologists Miles Joseph Berkeley and Christopher Edmund Broome who in 1883 formally described and named the fungus Sphaerella destructiva. That fungus is now known as Pseudopeziza medicaginis , the causal agent of common leaf spot of lucerne. It was one of over 300 fungi that were included in a 1880 paper co-written by the Reverend Julian Tenison-Woods and Frederick Bailey. At that time almost all of these fungi which had been collected in Australia were identified by overseas mycologists, particularly Berkeley and Broome. It can be argued that their 1880 paper was the first significant one published in Australia which focussed on fungi. Just a decade or so later Australian scientists, in particular Daniel McAlpine, were describing new fungal taxa on their own. Among the fungi recorded in a paper published in the 1880 Proceedings of the Linnaean Society of New South Wales was Sphaerella destructiva , now Pseudopeziza medicaginis , the cause of common leaf spot of lucerne. The paper, co-authored by the naturalist Reverend Julian Tenison-Woods and the Queensland Government Botanist Frederick Manson Bailey was the first known comprehensive list of Australian fungi published by Australian residents. It is a milestone in the evolution of mycology and plant pathology studies in Australia. Photograph by H. H. Baily. [ABSTRACT FROM AUTHOR]

Rhizoctonia solani is a soil-borne fungal species with worldwide distribution and poses serious threats to a wide range of economically important crops such as grain and forage crops. This pathogen has survival capabilities within plants and soil, giving rise to sclerotia and persisting for several months or years. Alfalfa (Medicago sativa) is the most widely grown and important forage crop in China and worldwide. The unique characteristics of alfalfa, such as excellent forage quality, ruminant desirability, and substantial biomass output, distinguish it from other fodder crops, and it is also known as the "Queen of Forages". However, the production of alfalfa is seriously affected by R. solani, with yield losses of 20% to 60% globally. This review firstly summarizes diseases such as seedling damping-off, root rot, crown rot, root cankers, stem cankers, blight, and stem rot caused by R. solani in alfalfa and the survival mechanism of this pathogen. The techniques for R. solani detection and quantification from plants and soils, as well as management through host resistance, cultural practices, fungicides, and biological control, were then overviewed. This review provides scientific knowledge to enable researchers to efficiently manage R. solani in alfalfa production. [ABSTRACT FROM AUTHOR]

Lucerne (Medicago sativa L.) forage production may be shifting towards the autumn–spring season, or in cooler environments, due to warmer and drier summers in Mediterranean Europe. This study aimed to evaluate the performance of lucerne cultivars with varying winter activity ratings (WAR) and hardiness in divergent environments of Greece: a cool highland versus a warm lowland. Highlywinter-active (HWA) cultivars were compared to semi-winter-active (SWA) ones for dry matter (DM) yield, seasonal harvest ratios, quality, and agronomic parameters. The SWA cultivars exhibited twice as many overwintered plants and higher summer yields, and were more productive (by 10.4%) in the cooler site. Conversely, HWA cultivars showed enhanced autumn yields and greater productivity (by 10.5%) in the warmer site. Notably, stability analyses revealed trade-offs between productivity and stability, with the most stable cultivar showing lower productivity (by 5.3–5.5%). Additionally, SWA cultivars exhibited higher crude protein content. Plant height and node number correlated with yields in the cool site but not in the warm, underscoring their effectiveness as indirect indicators in marginal environments. Outstanding temperature-specific cultivar responses fully justify the negative relation between winter activity and hardiness regarding productivity. This emphasizes the importance of matching cultivar winter activity and hardiness with specific microenvironments. Region-specific early screening could target the decoupling of the association between low winter activity and increased hardiness, enabling the optimization of cultivars for both traits, while interregional advanced line testing could capitalize on stability and resilience to address the challenges of climate change. [ABSTRACT FROM AUTHOR]

Understanding how a warmer and more variable climate will affect the productivity and stability of perennial forages in the establishment year is essential for determining forage management strategies resilient to the changing climate. We conducted an experiment from 2020–2021 and repeated it in 2021–2022 to determine how warmer and more variable temperatures affect yield of an alfalfa (Medicago sativa L.)–orchardgrass (Dactylis glomerata L.) mixture, as well as on the abundance of a winter annual weed, chickweed (Stellaria media), during the establishment year. We utilized open‐top chambers (OTCs) to manipulate air temperature in situ to impose three treatments: (1) constant warming (OTC present year‐round), (2) fluctuating warming (OTC present year‐round but temporarily removed prior to abrupt cold snaps), and (3) control (no OTC, ambient air temperature). OTCs increased air temperature within the alfalfa–orchardgrass canopy by an average of 0.45 and 0.72°C in Year 1 and Year 2, respectively; however, the effects of OTCs on air temperature varied temporally within and between years. Constant warming decreased alfalfa frost injury compared with the control, while the control and fluctuating warming resulted in similar levels of frost injury. Warmer temperature within the OTCs neither affected chickweed biomass nor the biomass of all other weed species. Both constant and fluctuating warming increased forage dry matter yield at the first harvest timepoint in the spring of both years, the period when preceding ambient air temperature tended to be cooler than other harvest timepoints. We found that the constant warming increased total forage yield (summed over the entire year) in Year 1, when precipitation was well above the 30‐year normal for our region, but decreased total forage yield in Year 2, when summer precipitation was substantially below the 30‐year normal. As global temperature continues to rise, yield loss of cool‐season perennial forages, such as alfalfa and orchardgrass, may be exacerbated in years without adequate precipitation. Therefore, exploring heat‐ and drought‐tolerant alternative forages, as well as management strategies that improve resilience to weather variability, will be increasingly important. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) are pivotal regulators of the abiotic stress responses in plants, yet their specific involvement in salt/alkali stress during alfalfa germination remains incompletely understood. Here, we subjected Zhongmu No.1 alfalfa (Medicago sativa L.) seeds to salt stress (20 mM NaCl and 20 mM Na2SO4 solutions) or alkali stress (5 mM NaHCO3 and 5 mM Na2CO3 solutions) treatments for 3 days, followed by total RNA extraction and RNA-seq analysis to delineate stress-responsive alfalfa lncRNAs. We identified 17,473 novel alfalfa lncRNAs, among which 101 and 123 were differentially expressed lncRNAs (DElncRNAs) under salt and alkali stress, respectively, compared to the control. Furthermore, we predicted 16 and 237 differentially expressed target genes regulated by DElncRNAs through cis/trans-regulatory mechanisms under salt or alkali stress, respectively. A functional enrichment analysis of DElncRNA target genes indicated that lncRNAs were implicated in the fatty acid metabolism pathway under salt stress, while they played a significant role in the phenylpropanoid and flavonoid biosynthesis pathway under alkali stress. Notably, lncRNAs were found to participate in the plant hormone signal transduction pathway, a common regulatory mechanism in both salt and alkali stress responses. These findings contribute to a deeper understanding of the mechanisms underlying alfalfa's response to salt and alkali stresses. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) is an economically important commodity in the Intermountain Western United States. A major concern for alfalfa producers in this region is the alfalfa weevil (Hypera postica Gyllenhal). Insecticide resistance development coupled with regulatory changes in pesticide use has resulted in renewed interest by producers in non-chemical control methods such as cultural control. One such cultural control method is early harvest, which consists of producers timing their harvests early in the season to decrease alfalfa weevil damage. This method is thought to be effective by exposing weevil larvae to adverse conditions before significant damage occurs. Still, early harvest can be difficult to employ because recommendations are often vague. To better understand how early harvest impacts both alfalfa weevils and their natural enemies and how producers are using this method across the Intermountain Western United States, we conducted a study in alfalfa production fields in Colorado, Montana, and Wyoming over three growing seasons. We determined that the timing of the initial alfalfa harvest spanned more than 1 month across fields, and alfalfa plant stage at harvest ranged from late vegetative to early bloom. Harvest was more impactful on reducing alfalfa weevil densities the earlier it was implemented. Removing windrows in a timely manner is likely useful to further decrease alfalfa weevil densities. Harvest timing was not associated with parasitism rates of alfalfa weevil, but higher parasitism rates were associated with lower post-harvest alfalfa weevil densities. This work has increased our understanding of early harvest in an on-farm setting and to improve recommendations for producers across the Intermountain Western United States. [ABSTRACT FROM AUTHOR]

Previous researchers utilized a single-rotor, screenless hammermill to improve forage utilization by dairy cows. They also established a surrogate measurement of animal response based on a relationship between processing level index (PLI) and in-situ digestibility. After a PLI of 60% was achieved, further processing resulted in diminished returns. However, to achieve a PLI of 60%, it required two to three processing passes through the single-rotor processor. In this research, a novel two-rotor processor was developed to achieve a PLI of 60% in a single pass through the machine. The final design featured two rotors with free-swinging blunt hammers. Material was transferred tangentially from the first to the second rotor. The first rotor was operated at tip speeds ranging from 66 to 110 m·s-1, and the second rotor tip speed could be adjusted to 1.10, 1.35, or 1.65 times greater than the first. Performance was evaluated by PLI, geometric mean particle size (GMPS), and gross specific energy (GSE). The processing of whole-plant corn at 51% w.b. moisture content and 50 mm length of cut resulted in PLI values of between 60% and 69% with a reduction in GMPS to 6 mm and a GSE requirement of 4.04 to 6.71 kWh Mg-1. Processing wilted alfalfa chopped at 35 mm length of cut and 67% w.b. moisture content increased PLI from 23% (chopped only) to 72% and reduced GMPS from 24 to 8 mm. The dual-rotor processor could process whole-plant corn and wilted alfalfa forages to the desired PLI of greater than 60% in a single pass. [ABSTRACT FROM AUTHOR]

Abstract Biochar is a promising solution to alleviate the negative impacts of salinity stress on agricultural production. Biochar derived from food waste effect was investigated on three plant species, Medicago sativa, Amaranthus caudatus, and Zea mays, under saline environments. The results showed that biochar improved significantly the height by 30%, fresh weight of shoot by 35% and root by 45% of all three species compared to control (saline soil without biochar adding), as well as enhanced their photosynthetic pigments and enzyme activities in soil. This positive effect varied significantly between the 3 plants highlighting the importance of the plant-biochar interactions. Thus, the application of biochar is a promising solution to enhance the growth, root morphology, and physiological characteristics of plants under salt-induced stress.

Abstract Background Auxin/induced-3-acetic acid (Aux/IAA) is an important plant hormone that affects plant growth and resistance to abiotic stresses. Drought stress is a vital factor in reducing plant biomass yield and production quality. Alfalfa (Medicago sativa L.) is the most widely planted leguminous forage and one of the most economically valuable crops in the world. Aux/IAA is one of the early responsive gene families of auxin, playing a crucial role in response to drought stress. However, the characteristics of the Aux/IAA gene family in alfalfa and its potential function in response to drought stress are still unknown. Result A total of 41 Aux/IAA gene members were identified in alfalfa genome. The physicochemical, peptide structure, secondary and tertiary structure analysis of proteins encoded by these genes revealed functional diversity of the MsIAA gene. A phylogenetic analysis classified the MsIAA genes into I-X classes in two subgroups. And according to the gene domain structure, these genes were classified into typical MsIAA and atypical MsIAA. Gene structure analysis showed that the MsIAA genes contained 1–4 related motifs, and except for the third chromosome without MsIAAs, they were all located on 7 chromosomes. The gene duplication analysis revealed that segmental duplication and tandem duplication greatly affected the amplification of the MsIAA genes. Analysis of the Ka/Ks ratio of duplicated MsAux/IAA genes suggested purification selection pressure was high and functional differences were limited. In addition, identification and classification of promoter cis-elements elucidated that MsIAA genes contained numerous elements associated to phytohormone response and abiotic stress response. The prediction protein–protein interaction network showed that there was a complex interaction between the MsAux/IAA genes. Gene expression profiles were tissue-specific, and MsAux/IAA had a broad response to both common abiotic stress (ABA, salt, drought and cold) and heavy metal stress (Al and Pb). Furthermore, the expression patterns analysis of 41 Aux/IAA genes by the quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed that Aux/IAA genes can act as positive or negative factors to regulate the drought resistance in alfalfa. Conclusion This study provides useful information for the alfalfa auxin signaling gene families and candidate evidence for further investigation on the role of Aux/IAA under drought stress. Future studies could further elucidate the functional mechanism of the MsIAA genes response to drought stress.

Biochar is a promising solution to alleviate the negative impacts of salinity stress on agricultural production. Biochar derived from food waste effect was investigated on three plant species, Medicago sativa, Amaranthus caudatus, and Zea mays, under saline environments. The results showed that biochar improved significantly the height by 30%, fresh weight of shoot by 35% and root by 45% of all three species compared to control (saline soil without biochar adding), as well as enhanced their photosynthetic pigments and enzyme activities in soil. This positive effect varied significantly between the 3 plants highlighting the importance of the plant-biochar interactions. Thus, the application of biochar is a promising solution to enhance the growth, root morphology, and physiological characteristics of plants under salt-induced stress. [ABSTRACT FROM AUTHOR]

Background: Auxin/induced-3-acetic acid (Aux/IAA) is an important plant hormone that affects plant growth and resistance to abiotic stresses. Drought stress is a vital factor in reducing plant biomass yield and production quality. Alfalfa (Medicago sativa L.) is the most widely planted leguminous forage and one of the most economically valuable crops in the world. Aux/IAA is one of the early responsive gene families of auxin, playing a crucial role in response to drought stress. However, the characteristics of the Aux/IAA gene family in alfalfa and its potential function in response to drought stress are still unknown. Result: A total of 41 Aux/IAA gene members were identified in alfalfa genome. The physicochemical, peptide structure, secondary and tertiary structure analysis of proteins encoded by these genes revealed functional diversity of the MsIAA gene. A phylogenetic analysis classified the MsIAA genes into I-X classes in two subgroups. And according to the gene domain structure, these genes were classified into typical MsIAA and atypical MsIAA. Gene structure analysis showed that the MsIAA genes contained 1–4 related motifs, and except for the third chromosome without MsIAAs, they were all located on 7 chromosomes. The gene duplication analysis revealed that segmental duplication and tandem duplication greatly affected the amplification of the MsIAA genes. Analysis of the Ka/Ks ratio of duplicated MsAux/IAA genes suggested purification selection pressure was high and functional differences were limited. In addition, identification and classification of promoter cis-elements elucidated that MsIAA genes contained numerous elements associated to phytohormone response and abiotic stress response. The prediction protein–protein interaction network showed that there was a complex interaction between the MsAux/IAA genes. Gene expression profiles were tissue-specific, and MsAux/IAA had a broad response to both common abiotic stress (ABA, salt, drought and cold) and heavy metal stress (Al and Pb). Furthermore, the expression patterns analysis of 41 Aux/IAA genes by the quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed that Aux/IAA genes can act as positive or negative factors to regulate the drought resistance in alfalfa. Conclusion: This study provides useful information for the alfalfa auxin signaling gene families and candidate evidence for further investigation on the role of Aux/IAA under drought stress. Future studies could further elucidate the functional mechanism of the MsIAA genes response to drought stress. [ABSTRACT FROM AUTHOR]

Floral bud growth influences seed yield and quality; however, the molecular mechanism underlying the development of floral buds in alfalfa (Medicago sativa) is still unclear. Here, we comprehensively analyzed the transcriptome and targeted metabolome across the early, mid, and late bud developmental stages (D1, D2, and D3) in alfalfa. The metabolomic results revealed that gibberellin (GA), auxin (IAA), cytokinin (CK), and jasmonic acid (JA) might play an essential role in the developmental stages of floral bud in alfalfa. Moreover, we identified some key genes associated with GA, IAA, CK, and JA biosynthesis, including CPS, KS, GA20ox, GA3ox, GA2ox, YUCCA6, amid, ALDH, IPT, CYP735A, LOX, AOC, OPR, MFP2, and JMT. Additionally, many candidate genes were detected in the GA, IAA, CK, and JA signaling pathways, including GID1, DELLA, TF, AUX1, AUX/IAA, ARF, GH3, SAUR, AHP, B-ARR, A-ARR, JAR1, JAZ, and MYC2. Furthermore, some TFs related to flower growth were screened in three groups, such as AP2/ERF-ERF, MYB, MADS-M-type, bHLH, NAC, WRKY, HSF, and LFY. The findings of this study revealed the potential mechanism of floral bud differentiation and development in alfalfa and established a theoretical foundation for improving the seed yield of alfalfa. [ABSTRACT FROM AUTHOR]

Studies documenting the consequences of harvest traffic in alfalfa production have addressed soil and plant growth parameters. One response was larger leaf/stem ratios in plants that were trafficked, which suggests higher quality. To fully understand how harvest traffic affects alfalfa quality a need for further analysis is warranted. Our objectives were to quantify differences in plant quality between trafficked and non-trafficked plants through 4 years of alfalfa production and to determine when these differences occur. The experimental units were furrow-irrigated raised beds with four harvests per year in Youngston clay loam soil in Fruita, Colorado. A John Deere 2280 swather and a John Deere 2955 tractor, driven over the alfalfa 7 days after swathing, were used to create four traffic treatments; plants that were never trafficked, plants trafficked only by the swather, plants trafficked only by the tractor, and plants trafficked by both the swather and the tractor. Quality was determined by measuring relative feed value, acid detergent fiber, neutral detergent fiber, and crude protein using near infrared reflectance spectroscopy. Alfalfa trafficked by the tractor had increased quality throughout the 4 years of production. [ABSTRACT FROM AUTHOR]

The environment in India is contaminated with polycyclic aromatic hydrocarbons (PAHs) due to the occurrence of large anthropogenic activities, i.e., fuel combustion, mineral roasting, and biomass burning. Hence, 13 toxic PAHs were detected: phenanthrene, anthracene, fluoranthene, pyrene, and benz(a) anthracene, ben-zo; (b) fluoranthene, benzo(k) fluoranthene, benzo(a) pyrene, benzo(ghi)perylene, dibenz (ah) anthracene, indeno1,2,3-(cd) pyrene, coronene and coronene in the environment (i.e., ambient particulate matter, road dust, sludge, and sewage) of the most industrialized area. Pollutants such as heavy metals and polycyclic aromatic hydrocarbons co-contaminate the soil and pose a significant hazard to the ecosystem because these pollutants are harmful to both humans and the environment. Phytoremediation is an economical plant-based natural approach for soil clean-up that has no negative impact on ecosystems. The aim of this study was to investigate the effects of pyrene (500 mg kg−1), Zn (150 mg kg−1), Pb (150 mg kg−1), and Cd (150 mg kg−1) alone and in combination on the phytoextraction efficiency of Medicago sativa growing in contaminated soil. Plant biomass, biochemical activities, translocation factors, accumulation of heavy metals, and pyrene removal were determined. After 60 days of planting, compared with those of the control plants, the growth parameters, biomass, and chlorophyll content of the M. sativa plants were significantly lower, and the reactive oxygen species activity, such as proline and polyphenol content and metallothionein protein content, was markedly greater in the pyrene and heavy metal-polluted soils. Furthermore, the combined toxicity of pyrene and all three metals on M. sativa growth and biochemical parameters was significantly greater than that of pyrene, Zn, Pb, or Cd alone, indicating the synergistic effect of pyrene and heavy metals on cytotoxicity. Pyrene stress increased Cd accumulation in M. sativa. After pyrene exposure alone or in combination with Zn-pyrene, a greater pyrene removal rate (85.5–81.44%) was observed than that in Pb-pyrene, Cd-pyrene, and Zn-Pb-Cd-pyrene polluted soils (62.78–71.27%), indicating that zinc can enhance the removal of pyrene from contaminated soil. The resulting hypotheses demonstrated that Medicago sativa can be used as a promising phytoremediation agent for co-contaminated soil. [ABSTRACT FROM AUTHOR]

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-associated proteins are a class of transmembrane proteins involved in intracellular trafficking pathways. However, the functions of many SNARE domain-containing proteins remain unclear. We have previously identified a SNARE-associated gene in alfalfa (Medicago sativa) KILLING ME SLOWLY1 (MsKMS1), which is involved in various abiotic stresses. In this study, we investigated the function of MsKMS1 in the seed germination of transgenic tobacco (Nicotiana tabacum). Phylogenetic analysis showed that MsKMS1 was homologous to the SNARE-associated or MAPR component-related proteins of other plants. Germination assays revealed that MsKMS1 negatively regulated seed germination under normal, D-mannitol and abscisic acid-induced stress conditions, yet MsKMS1 -overexpression could confer enhanced heat tolerance in transgenic tobacco. The suppressive effect on germination in MsKMS1 -overexpression lines was associated with higher abscisic acid and salicylic acid contents in seeds. This was accompanied by the upregulation of abscisic acid biosynthetic genes (ZEP and NCED) and the downregulation of gibberellin biosynthetic genes (GA20ox2 and GA20ox3). Taken together, these results suggested that MsKMS1 negatively regulated seed germination by increasing abscisic acid and salicylic acid contents through the expression of genes related to abscisic acid and gibberellin biosynthesis. In addition, MsKMS1 could improve heat tolerance during the germination of transgenic tobacco seeds. Plants, as sessile, have developed a series of mechanisms to adapt the environmental stresses, which was regulated by a cascade of genes and transcriptional factors. Here, we analysed the gene function of MsKMS1 in seed germination of transgenic tobacco (Nicotiana tabacum) under abiotic stress conditions and found that MsKMS1 negatively affected seed germination by increasing ABA and salicylic acid contents, regulating the expression of genes related to ABA and GA biosynthesis. [ABSTRACT FROM AUTHOR]

El cuy en el Ecuador es sinónimo de nuestra gastronomía ancestral y de tradición cultural, pero por la alimentación rica en L-canavanina (alfalfa) que reciben; podría limitar su consumo en pacientes con trombocitopenia inmune primaria (PTI). La ingesta de alfalfa en humanos puede propiciar insuficiencia renal y síndrome lupus-like. El centro de Lupus John Hopkins recomiendan evitarla en la dieta de los pacientes con Lupus Eritematoso Sistémico (LES), al agravar la inflamación por estimulación de la actividad inmune (flares). Presentamos dos casos de pacientes con PTI vinculados con la ingesta de cuy. ¿Es probable?

Abstract Understanding how a warmer and more variable climate will affect the productivity and stability of perennial forages in the establishment year is essential for determining forage management strategies resilient to the changing climate. We conducted an experiment from 2020–2021 and repeated it in 2021–2022 to determine how warmer and more variable temperatures affect yield of an alfalfa (Medicago sativa L.)–orchardgrass (Dactylis glomerata L.) mixture, as well as on the abundance of a winter annual weed, chickweed (Stellaria media), during the establishment year. We utilized open‐top chambers (OTCs) to manipulate air temperature in situ to impose three treatments: (1) constant warming (OTC present year‐round), (2) fluctuating warming (OTC present year‐round but temporarily removed prior to abrupt cold snaps), and (3) control (no OTC, ambient air temperature). OTCs increased air temperature within the alfalfa–orchardgrass canopy by an average of 0.45 and 0.72°C in Year 1 and Year 2, respectively; however, the effects of OTCs on air temperature varied temporally within and between years. Constant warming decreased alfalfa frost injury compared with the control, while the control and fluctuating warming resulted in similar levels of frost injury. Warmer temperature within the OTCs neither affected chickweed biomass nor the biomass of all other weed species. Both constant and fluctuating warming increased forage dry matter yield at the first harvest timepoint in the spring of both years, the period when preceding ambient air temperature tended to be cooler than other harvest timepoints. We found that the constant warming increased total forage yield (summed over the entire year) in Year 1, when precipitation was well above the 30‐year normal for our region, but decreased total forage yield in Year 2, when summer precipitation was substantially below the 30‐year normal. As global temperature continues to rise, yield loss of cool‐season perennial forages, such as alfalfa and orchardgrass, may be exacerbated in years without adequate precipitation. Therefore, exploring heat‐ and drought‐tolerant alternative forages, as well as management strategies that improve resilience to weather variability, will be increasingly important.

Insect pollinators affect pollen transfer dynamics, with consequences for pollen movement and the genetic structure of plant populations. Pollen transfer dynamics has not been previously examined in flowers with a tripping mechanism. Here we examine whether pollen accumulated on a bee’s body increases with the number of Medicago sativa L. flowers tripped by Bombus impatiens Cresson during a foraging bout, while controlling for bee body size and number of visited flowers. In a second experiment, we determine whether the number of revisits to a tripped flower increases pollen deposition onto the stigmas. We set up three M. sativa plants with a controlled number of racemes in a greenhouse room, and followed individual bees as they foraged, recording each plant, raceme, and flower visited. For pollen accumulated, we collected bees at the end of their foraging bout and counted pollen grains on their body. For pollen deposition, we collected flowers with between 0 and 6 revisits and counted the pollen grains on the stigmas. The number of pollen grains on a bee’s body increased with the number of flowers tripped in a foraging bout, but was not affected by the number of flowers visited or the size of individual bees. The number of pollen grains deposited on a stigma did not increase with the number of revisits to a tripped flower. This latter result contrasts with plants without a tripping mechanism where the number of visits increases pollen deposition and seed set. Tripping affects pollen transfer dynamics and we discuss how its effect may vary with the mode of tripping.

Objective: Medicago sativa (M. sativa) has been traditionally used for treating anemia; therefore, M. sativa hydro-ethanolic extract therapeutic effects against cyclophosphamide (CP) -induced hematologic and liver toxicity were examined. Materials and Methods: Thirty male Wistar rats were randomly divided to control (saline); CP (100 mg/kg, day 1-3, subcutaneously); CP+ M. sativa 200 mg/kg (MS 200); CP+ M. sativa 400 mg/kg (MS 400); CP+ dexamethasone (0.1 mg/kg), (all groups n=6). Treated animals received M. sativa or dexamethasone by gavage from days 7-14. On days 0, 7, and 14, hematologic parameters, and on the 14th day, serum and liver tissue oxidative stress markers including nitric oxide, malondialdehyde (MDA) and total thiol levels, superoxide dismutase (SOD) and catalase (CAT) activities, serum lipids, and liver enzymes were measured.Results: Animal weight, platelet, white blood cells, and red blood cells counts, hemoglobin and hematocrit as well as thiol, SOD, and CAT activities in serum and liver tissue were significantly reduced, but serum nitric oxide, MDA, total cholesterol, triglycerides, low-density lipoproteins levels, and liver enzymes were increased in the CP group compared to the control group (p

Abstract Background Odorant-binding proteins (OBPs) are essential in insect’s daily behaviors mediated by olfactory perception. Megachile saussurei Radoszkowski (Hymenoptera, Megachilidae) is a principal insect pollinating alfalfa (Medicago sativa) in Northwestern China. The olfactory function have been less conducted, which provides a lot of possibilities for our research. Results Our results showed that 20 OBPs were identified in total. Multiple sequence alignment analysis indicated MsauOBPs were highly conserved with a 6-cysteine motif pattern and all belonged to the classic subfamily, coding 113-196 amino acids and sharing 41.32%-99.12% amino acid identity with known OBPs of other bees. Phylogenetic analysis indicated there were certain homologies existed among MsauOBPs and most sequences were clustered with that of Osmia cornuta (Hymenoptera, Megachilidae). Expression analysis showed the identified OBPs were mostly enriched in antennae instead of other four body parts, especially the MsauOBP2, MsauOBP3, MsauOBP4, MsauOBP8, MsauOBP11 and MsauOBP17, in which the MsauOBP2, MsauOBP4 and MsauOBP8 presented obvious tissue-biased expression pattern. Molecular docking results indicated MsauOBP4 might be the most significant protein in recognizing alfalfa flower volatile 3-Octanone, while MsauOBP13 might be the most crucial protein identifying (Z)-3-hexenyl acetate. It was also found the lysine was a momentous hydrophilic amino acid in docking simulations. Conclusion In this study, we identified and analyzed 20 OBPs of M. saussurei. The certain homology existed among these OBPs, while some degree of divergence could also be noticed, indicating the complex functions that different MsauOBPs performed. Besides, the M. saussurei and Osmia cornuta were very likely to share similar physiological functions as most of their OBPs were clustered together. MsauOBP4 might be the key protein in recognizing 3-Octanone, while MsauOBP13 might be the key protein in binding (Z)-3-hexenyl acetate. These two proteins might contribute to the alfalfa-locating during the pollination process. The relevant results may help determine the highly specific and effective attractants for M. saussurei in alfalfa pollination and reveal the molecular mechanism of odor-evoked pollinating behavior between these two species.

Leaves are the most significant parts of forage crops such as alfalfa. Senescence is the terminal stage of leaf development and is controlled by an integrated myriad of endogenous signals and environmental stimuli. WRKY transcription factors (TFs) play essential roles in regulating leaf senescence; however, only a few studies on the analysis and identification of the WRKY TF family in Medicago Sativa have been reported. In this study, we identified 198 WRKY family members from the alfalfa (M. sativa L.) cultivar ’XinjiangDaye’ using phylogenetic analysis and categorized them into three subfamilies, Groups I, II, and III, based on their structural characteristics. Group II members were further divided into five subclasses. In addition, several hormone- and stress-related cis-acting elements were identified in the promoter regions of MsWRKYs. Furthermore, 14 aging-related MsWRKYs genes from a previous transcriptome in our laboratory were selected for RT-qPCR validation of their expression patterns, and subsequently cloned for overexpression examination. Finally, MsWRKY5, MsWRKY66, MsWRKY92, and MsWRKY141 were confirmed to cause leaf yellowing in Nicotiana benthaminana using a transient expression system. Our findings lay a groundwork for further studies on the mechanism of M. sativa leaf aging and for the creation of new germplasm resources.

This study investigated using Hedychium gardnerianum, an invasive plant, combined with dehydrated alfalfa (Medicago sativa) with varying percentages of alfalfa: control (0% alfalfa), T1 (10%), T2 (20%), T3 (30%), and T4 (40%), to enhance nutritional parameters in silage for ruminants in the Azores. Samples were vacuum-sealed and stored for 45 days at room temperature to promote anaerobic fermentation. Chemical analysis showed that alfalfa addition increased dry matter (DM) from 12.33% to 48.04% and crude protein from 11.34% DM to 24.63% DM. Insoluble fiber levels decreased, enhancing digestibility. In vitro fermentation indicated higher gas production in 40% alfalfa silage, suggesting faster fermentation. In conclusion, incorporating dehydrated alfalfa into Hedychium gardnerianum silage particularly enhances nutritional quality and digestibility. This approach offers a practical solution for ruminant feed in the Azores, particularly during feed scarcity periods.

Soil salinization is a common environmental problem that seriously threatens crop yield and food security, especially through its impact on seed germination. Nanoparticle priming, an emerging seed treatment method, is receiving increasing attention in improving crop yield and stress resistance. This study used alfalfa seeds as materials to explore the potential benefits of cerium oxide nanoparticle (CeO2NP) priming to promote seed germination and improve salt tolerance. CeO2NPs at concentrations up to 500 mg/L were able to significantly alleviate salt stress in alfalfa seeds (200 mM), with 50 mg/L of CeO2NP having the best effect, significantly (P< 0.05) increasing germination potential (from 4.0% to 51.3%), germination rate (from 10.0% to 62.7%), root length (from 8.3 cm to 23.1 cm), and seedling length (from 9.8 cm to 13.7 cm). Priming treatment significantly (P< 0.05) increased seed water absorption by removing seed hardness and also reducing abscisic acid and jasmonic acid contents to relieve seed dormancy. CeO2NP priming increased α-amylase activity and osmoregulatory substance level, decreased reactive oxygen species and malonaldehyde contents and relative conductivity, and increased catalase enzyme activity. Seed priming regulated carotenoid, zeatin, and plant hormone signal transduction pathways, among other metabolic pathways, while CeO2NP priming additionally promoted the enrichment of α-linolenic acid and diterpenoid hormone metabolic pathways under salt stress. In addition, CeO2NPs enhanced α-amylase activity (by 6.55%) in vitro. The optimal tested concentration (50 mg/L) of CeO2NPs was able to improve the seed vigor, enhance the activity of α-amylase, regulate the osmotic level and endogenous hormone levels, and improve the salt tolerance of alfalfa seeds. This study demonstrates the efficacy of a simple seed treatment strategy that can improve crop stress resistance, which is of great importance for reducing agricultural costs and promoting sustainable agricultural development. [ABSTRACT FROM AUTHOR]

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

Indigenous chicken production consists of an array of activities important to smallholder poultry farmers in Africa. One of the many factors influencing their production and threating the local food security is in the area of nutrition, particularly, that related to protein supplementation. The available feed resources to farmers are not enough to sustain the productivity of the chickens. Hence, the chickens’ diets often require nutritional supplementation. There is therefore an urgent need for the validation of locally grown feed ingredients to improve the sustainability of poultry production in sub-Saharan Africa. A dietary ingredient that may be used in the diets of chickens is lucerne (Medicago sativa), which is also known as alfalfa. In South Africa, lucerne is the most cultivated forage legume and approximately 1.3 million metric tonnes of lucerne are produced per year. Lucerne has high nutritional value, as it is a source of protein, amino acids, vitamins, and fatty acids. The potential of lucerne as a feed resource for indigenous chickens should, therefore, be investigated so that strategies to improve the nutrition of such chickens can be developed. The purpose of this review was to highlight lucerne as a potential dietary ingredient for indigenous chickens and discuss its effects on the productivity of broilers, egg-layers, and dual-purpose chickens. [ABSTRACT FROM AUTHOR]

We investigated the effects of Alfalfa (Medicago sativa L.) and Bermudagrass (Cynodon dactylon L.) extracts concentrations on callus induction and rye (Secale cereale L.) embryo regeneration and growth of rye seedlings in both greenhouse and in-vitro conditions. Immature rye embryos were used as explants for generating callus, and water extracts of alfalfa and bermudagrass were added to the environment with a concentration of 5, 10, 15, 20, 25 and 30 %. This extract was also used in the regeneration period. The use of 25 and 30 % alfalfa extracts stopped callus induction and its components. The application of bermudagrass extract with a concentration of 30 %, reduced rye callus induction to 50 %. Alfalfa extract concentrations of 20 and 25 %, respectively caused a reduction of 35 and 75 % in callus fresh weight in comparison with the control conditions. Applying 25 and 30 % concentrations of bermudagrass extract led to a reduction of 68 and 80 % respectively. Alfalfa extracts with 15, 20 and 25 % concentrations caused reductions of 40, 63 and 69 % in rooting. The 30 % concentration of M. sativa extract stopped rooting in rye callus. Concentrations of 15, 20, 25 and 30 % caused reductions of 31, 50, 55 and 81 % in the shooting of rye callus, respectively. The results indicate that alfalfa extract had greater effect on rooting, and the bermudagrass extracts had greater effect on shooting. The analysis of the results shows that Alfalfa and bermudagrass extracts have a strong allelopathic effect, which can be used in the production of herbicides of natural origin. [ABSTRACT FROM AUTHOR]

Objective: Medicago sativa (M. sativa) has been traditionally used for treating anemia; therefore, M. sativa hydro-ethanolic extract therapeutic effects against cyclophosphamide (CP) - induced hematologic and liver toxicity were examined. Materials and Methods: Thirty male Wistar rats were randomly divided to control (saline); CP (100 mg/kg, day 1-3, subcutaneously); CP+ M. sativa 200 mg/kg (MS 200); CP+ M. sativa 400 mg/kg (MS 400); CP+ dexamethasone (0.1 mg/kg), (all groups n=6). Treated animals received M. sativa or dexamethasone by gavage from days 7-14. On days 0, 7, and 14, hematologic parameters, and on the 14th day, serum and liver tissue oxidative stress markers including nitric oxide, malondialdehyde (MDA) and total thiol levels, superoxide dismutase (SOD) and catalase (CAT) activities, serum lipids, and liver enzymes were measured. Results: Animal weight, platelet, white blood cells, and red blood cells counts, hemoglobin and hematocrit as well as thiol, SOD, and CAT activities in serum and liver tissue were significantly reduced, but serum nitric oxide, MDA, total cholesterol, triglycerides, low-density lipoproteins levels, and liver enzymes were increased in the CP group compared to the control group (p<0.05 to p<0.001). Administering M. sativa extract (400 mg/kg) significantly enhanced platelet count, and SOD and CAT activities and inhibited all of the CP toxic effects, while dexamethasone improved platelet count and oxidative stress markers compared to the CP group (p<0.05 to p<0.001). Conclusion: The extract of M. sativa (400 mg/kg) showed therapeutic effects against the CP-induced myelosuppression and thrombocytopenia and improved oxidative stress markers which were comparable to the effect of dexamethasone. [ABSTRACT FROM AUTHOR]

Alfalfa Paraphoma root rot (APRR) (Paraphoma radicina) is a recently described alfalfa disease widely distributed in China, first reported in 2020. So far, the resistance levels of 30 alfalfacultivars to APRR have been characterized; however, the resistance mechanisms among these cultivars remain unknown. In the present study, the alfalfa resistance mechanisms against APRR were investigated by studying the ditference of P. radicina infecting susceptible (Gibraltar) and resistant (Magnum II) alfalfa cultivars under the light microscope and scanning electronic microscope. The conidial germination and germ tube growth in the root exudates of different resistant cultivars were also compared. The results revealed that conidial germination, germ tube development, and P. radicina penetration into root tissues of resistant plants were delayed. In susceptible and resistant cultivars, P. radicina infected roots by penetrating epidermal cells and the intercellular space between epidermal cells. During the infection process, germ tubes penetrated the root surface directly or formed appressoria. However, the penetration percentage on the susceptible cultivar was significantly higher than on the resistant cultivar, irrespective of the infection route. Moreover, disintegrated conidia and germ tubes were observed on resistant cultivar roots at 48 h postinoculation. The conidial germination and germ tube growth in root exudates of susceptible cultivars were significantly higher than in resistant cultivars. The current findings implied that the alfalfa resistance mechanism might be related to root exudates. These findings could provide insights into the alfalfa resistance mechanism following P. radicina infection. [ABSTRACT FROM AUTHOR]

The objective of present study was to assess seed yield and yield related traits of alfalfa clonal progenies. The phenotypic variation of traits within progenies was also determined. Eleven alfalfa clonal progenies of native origin were object of investigation. The experiment was carried out at the Experimental field of the IASS Obraztsov Chiflik, Rousse for three-year period. The traits plant seed yield (PSY), plant height (PH), generative stem number (GSN), inflorescence number (INP), pod number (PNI), seed number (SNP) and 1000-seed weight (TSW) were evaluated. There were statistically significant differences among clonal progenies for all studied traits. Five progenies (PM30, JM13, GM27, SL83 and PM18) were identified as very valuable genetic source with potential for developing high seed yielding varieties. There was established PSY, PNI and SNP exhibited moderate to high phenotypic variability within progenies, while PH and TSW expressed low variability. Data confirm possibility of used the traits number of pods per inflorescence and number of seeds per pod as criteria of germplasm selection for seed yield improvement. [ABSTRACT FROM AUTHOR]

Plant mitochondria are crucial for species evolution, phylogenetics, classification, and identification as maternal genetic material. However, the presence of numerous repetitive sequences, complex structures, and a low number of genes in the mitochondrial genome has hindered its complete assembly and related research endeavors. In this study, we assembled two mitochondrial genomes of alfalfa varieties of Zhongmu No.1 (299,123 bp) and Zhongmu No.4 (306,983 bp), based on a combination of PacBio, Illumina, and Hi-C sequences. The comparison of genome assemblies revealed that the same number of mitochondrial genes, including thirty-three protein-coding genes, sixteen tRNA genes, and three rRNA genes existed in the two varieties. Additionally, large fragments of repetitive sequences were found underlying frequent mitochondrial recombination events. We observed extensive transfer of mitochondrial fragments into the nuclear genome of Zhongmu No.4. Analysis of the cox1 and rrn18s genes in 35 Medicago accessions revealed the presence of population-level deletions and substitutions in the rrn18s gene. We propose that mitochondrial structural reorganizations may contribute to alfalfa evolution. [ABSTRACT FROM AUTHOR]