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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: 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 FROM AUTHOR]

Atrazine, a commonly employed herbicide for corn production, can leave residues in soil, resulting in photosynthetic toxicity and impeding growth in subsequent alfalfa (Medicago sativa L.) crops within alfalfa-corn rotation systems. The molecular regulatory mechanisms by which atrazine affects alfalfa growth and development, particularly its impact on the microbial communities of the alfalfa rhizosphere, are not well understood. This study carried out field experiments to explore the influence of atrazine stress on the biomass, chlorophyll content, antioxidant system, and rhizosphere microbial communities of the atrazine-sensitive alfalfa variety WL- 363 and the atrazine-resistant variety JN5010. The results revealed that atrazine significantly reduced WL-363 growth, decreasing plant height by 8.58cm and root length by 5.42cm (p <0.05). Conversely, JN5010 showed minimal reductions, with decreases of 1.96cm in height and 1.26cm in root length. Chlorophyll content in WL-363 decreased by 35% under atrazine stress, while in JN5010, it was reduced by only 10%. Reactive oxygen species (ROS) accumulation increased by 60% in WL- 363, compared to a 20% increase in JN5010 (p <0.05 for both). Antioxidant enzyme activities, such as superoxide dismutase (SOD) and catalase (CAT), were significantly elevated in JN5010 (p <0.05), suggesting a more robust defense mechanism. Although the predominant bacterial and fungal abundances in rhizosphere soils remained generally unchanged under atrazine stress, specific microbial groups exhibited variable responses. Notably, Promicromonospora abundance declined in WL-363 but increased in JN5010. FAPROTAX functional predictions indicated shifts in the abundance of microorganisms associated with pesticide degradation, resistance, and microbial structure reconstruction under atrazine stress, displaying different patterns between the two varieties. This study provides insights into how atrazine residues affect alfalfa rhizosphere microorganisms and identifies differential microbial responses to atrazine stress, offering valuable reference data for screening and identifying atrazine-degrading bacteria. [ABSTRACT FROM AUTHOR]

Flower development is a crucial and complex process in the reproductive stage of plants, which involves the interaction of multiple endogenous signals and environmental factors. However, regulatory mechanism of flower development was unknown in alfalfa (Medicago sativa). In this study, the three stages of flower development of 'M. sativa cv. Gannong No. 5' (G5) and its early flowering and multi flowering mutant (MG5) were comparatively analyzed by transcriptomics. The results showed that compared with late bud stage (S1), 14287 and 8351 differentially expressed genes (DEGs) were identified at early flower stage (S2) in G5 and MG5, and 19941 and 19469 DEGs were identified at late flower stage (S3). Compared with S2, 9574 and 10870 DEGs were identified at S3 in G5 and MG5, respectively. Venn analysis revealed that 547 DEGs were identified among the three comparison groups. KEGG pathway enrichment analysis showed that these genes were involved in the development of alfalfa flowers through redox pathways and plant hormone signaling pathways. Key candidate genes including SnRK2, BSK, GID1, DELLA and CRE1, for regulating the development from buds to mature flowers in alfalfa were screened. In addition, differential expression of transcription factors such as MYB, AP2, bHLH, C2C2, MADS-box, NAC, bZIP, B3 and AUX/IAA also played an important role in this process. The results laid a theoretical foundation for studying the molecular mechanisms of the development process from buds to mature flowers in alfalfa. [ABSTRACT FROM AUTHOR]

Background: The mitogen-activated protein kinase (MAPK) cascade is crucial cell signal transduction mechanism that plays an important role in plant growth and development, metabolism, and stress responses. The MAPK cascade includes three protein kinases, MAPK, MAPKK, and MAPKKK. The three protein kinases mediate signaling to downstream response molecules by sequential phosphorylation. The MAPK gene family has been identified and analyzed in many plants, however it has not been investigated in alfalfa. Results: In this study, Medicago sativa MAPK genes (referred to as MsMAPKs) were identified in the tetraploid alfalfa genome. Eighty MsMAPKs were divided into four groups, with eight in group A, 21 in group B, 21 in group C and 30 in group D. Analysis of the basic structures of the MsMAPKs revealed presence of a conserved TXY motif. Groups A, B and C contained a TEY motif, while group D contained a TDY motif. RNA-seq analysis revealed tissue-specificity of two MsMAPKs and tissue-wide expression of 35 MsMAPKs. Further analysis identified MsMAPK members responsive to drought, salt, and cold stress conditions. Two MsMAPKs (MsMAPK70 and MsMAPK75) responds to salt and cold stresses; two MsMAPKs (MsMAPK60 and MsMAPK73) responds to cold and drought stresses; four MsMAPKs (MsMAPK1, MsMAPK33, MsMAPK64 and MsMAPK71) responds to salt and drought stresses; and two MsMAPKs (MsMAPK5 and MsMAPK7) responded to all three stresses. Conclusion: This study comprehensively identified and analysed the alfalfa MAPK gene family. Candidate genes related to abiotic stresses were screened by analysing the RNA-seq data. The results provide key information for further analysis of alfalfa MAPK gene functions and improvement of stress tolerance. [ABSTRACT FROM AUTHOR]

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 FROM AUTHOR]

Leaves are a key forage part for livestock, and the aging of leaves affects forage biomass and quality. Preventing or delaying premature leaf senescence leads to an increase in pasture biomass accumulation and an improvement in alfalfa quality. NAC transcription factors have been reported to affect plant growth and abiotic stress responses. In this study, 48 NAC genes potentially associated with leaf senescence were identified in alfalfa under dark or salt stress conditions. A phylogenetic analysis divided MsNACs into six subgroups based on similar gene structure and conserved motif. These MsNACs were unevenly distributed in 26 alfalfa chromosomes. The results of the collinearity analysis show that all of the MsNACs were involved in gene duplication. Some cis-acting elements related to hormones and stress were screened in the 2-kb promoter regions of MsNACs. Nine of the MsNAC genes were subjected to qRT-PCR to quantify their expression and Agrobacterium-mediated transient expression to verify their functions. The results indicate that Ms.gene031485, Ms.gene032313, Ms.gene08494, and Ms.gene77666 might be key NAC genes involved in alfalfa leaf senescence. Our findings extend the understanding of the regulatory function of MsNACs in leaf senescence. [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]

Sowing date and soil fertility are very important factors in the overwintering and production performance of alfalfa (Medicago sativa L.), yet there's a knowledge gap in knowledge on how late-seeded alfalfa responds to phosphorus (P) fertilization. A field study was conducted in Inner Mongolia from 2020 to 2022 using a split-plot design. The main plots consisted of five sowing dates (31 July, 8, 16, and 24 August, and 1 September), while the subplots involved five P application rates (0, 40, 70, 100, and 130 kg P2O5 ha−1). Throughout the growing seasons, the overwintering rate, root traits, forage yield, and yield components were measured. The results revealed a consistent decrease in overwintering ability and productivity with the delayed sowing. This reduction in overwintering rate was mainly due to diminished root traits, while the decrease in forage yield was largely associated with a reduction in plants per square meter. However, P fertilizer application to late-seeded alfalfa demonstrated potential in enhancing the diameter of both the crown and taproot, thus strengthening the root system and improving the overwintering rate, the rate of increase ranges from 11.6 to 49%. This adjustment could also improve the shoots per square meter and mass per shoot, increasing by 9.4–31.3% and 15.0–27.1% respectively in 2 years, which can offset the decline in forage yield caused by late sowing and might even increase the forage yield. Regression and path analysis indicated that alfalfa forage yield is primarily affected by mass per shoot rather than shoots per square meter. This study recommended that the sowing of alfalfa in similar regions of Inner Mongolia should not be later than mid-August. Moreover, applying P fertilizer (P2O5) at 70.6–85.9 kg ha−1 can enhance the forage yield and persistence of late-seeded alfalfa. Therefore, appropriate late sowing combined with the application of P fertilizer can be used as an efficient cultivation strategy for alfalfa cultivation after a short-season crop harvest in arid and cold regions. [ABSTRACT FROM AUTHOR]

Alfalfa is one of the most important and the most cultivated crop due to its high nutritive quality and yield, but adaptation of alfalfa genotypes differ in terms of mobile aluminium stress in the soil. The aim of this study was to evaluate the tolerance to mobile Al concentrations in the laboratory and in the naturally acidic soil and select the promising genotypes based on agro-biological traits. In 2019, a laboratory experiment was conducted at the Institute of Agriculture of LAMMC. The experiment in the acidic soil with different mobile Al concentrations was conducted at the Vėžaičiai Branch of LAMMC. In 2020, the crops of alfalfa genotypes (11 cultivars and 3 populations) were established on Balthygleyic Dystric Retisol. The agro-biological traits were assessed during the 2021–2022 season. The tolerance index of hypocotyls and roots was evaluated using the filter-based screening method at different AlCl3 (0.0–64 mM) concentrations. The study results of the filter-based screening method showed that the genotype Žydrūnė, Malvina, Jõgeva 118, Skriveru, and 3130 were the most tolerant ones and the hypocotyl tolerance index of these genotypes was higher compared to medium tolerant genotypes Birutė, PGR12489, Europe and AJ2024 at 8, 16, 32 and 64 mM AlCl3 concentrations. The hypocotyl and root tolerance index of medium tolerant genotypes was higher compared to a sensitive genotype PGR10249 at 8 and 16 mM AlCl3. The study of cluster analysis with mobile Al 0.0–65.0 mg kg-1 showed that the genotypes Žydrūnė, Europe, AJ2024 and 3130 were the best in terms of wintering and spring regrowth, the cultivar Malvina had the best value of wintering, height before flowering and stem number, the cultivar Birutė had the best value of spring regrowth, height before flowering and seed yield, and the cultivar Skriveru had the best value of spring regrowth, height before flowering, stem number and seed yield. [ABSTRACT FROM AUTHOR]

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. [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 feed components contributes to variation and uncertainty of diets delivered to dairy cows. Forages often have a high inclusion rate (50% to 70% of DM fed) and variable composition, and thus are an important contributor to nutrient variability of delivered diets. Our objective was to quantify the variation and identify the main sources of variability in corn silage and alfalfa-grass haylage composition at harvest (fresh forage) and feed-out (fermented forage) on New York dairy farms. Corn silage and alfalfa-grass haylage were sampled on 8 New York commercial dairy farms during harvest in the summer and fall of 2020 and during their subsequent feed-out in the winter and spring of 2021. At harvest, a composite sample of fresh chopped forage of every 8-ha section of individual fields was collected from piles delivered for silo filling. During a 16-wk feed-out period, 2 independent samples of each forage were collected 3 times per week. The fields of origin of each forage sample during feed-out were identified and recorded using silo maps created at filling. A mixed-model analysis quantified the variance of corn silage DM, NDF, and starch and haylage DM, NDF, and CP content. Fixed effects included soil type, weather conditions, and management practices during harvest and feed-out, and random effects were farm, silo unit, field, and day. At harvest, between-farm variability was the largest source of variation for both corn silage and haylage, but within-farm sources of variation exceeded farm-to-farm variation for haylage at feed-out. At feed-out, haylage DM and NDF content had higher within-farm variability than corn silage. In contrast, corn silage starch showed higher within-farm variation at feed-out than haylage CP content. For DM content at feed-out, day-to-day variation was the most relevant source of within-farm variation for both forages. However, for the nutrient components at feed-out (NDF and CP for haylage; NDF and starch for corn silage) silo-to-silo variation was the largest source of variability. Weather conditions systematically explained a proportion of the farm-to-farm variability for both forages at harvest and feed-out. We concluded that because of the high farm-to-farm variation, corn silage and haylage must be sampled on individual farms. We also concluded that due to the high silo-to-silo variability, and the still significant day-to-day and field-to-field variability within-farm, corn silage and haylage should be sampled within individual silos to better capture changes in forage components at feed-out. [ABSTRACT FROM AUTHOR]

Sustainable management of potassium nutrition in alfalfa crop production is one of the major key factors for achieving optimum seed and biomass yields. An inappropriate supply of mineral potassium nutrition in alfalfa production could result in a decrease in biomass and grain yield production, leading to luxury consumption with cost implications. Alfalfa (Medicago sativa L.) is a perennial leguminous forage crop known for its high protein content, nutritive value, biomass yield production, soil-improving abilities, and livestock feed. Potassium nutrition plays a crucial role in alfalfa production by influencing several physiological processes essential for biomass yield, growth, development, photosynthesis, nutrient uptake, and stress tolerance. Although several studies have been conducted regarding the role of potassium nutrition in agriculture productivity, only limited research has focused on crop-specific impacts. Therefore, this paper reviews (i) the significant role potassium nutrition plays in alfalfa production along with its implications for quality, yield, growth, and resistance to abiotic stress; (ii) the factors affecting the availability, absorption, and transport of potassium; (iii) the source of potassium and the consequences of inadequate availability; and (iv) highlights some strategies for mitigating potassium nutrient deficiency to optimize alfalfa productivity and sustainability in agricultural systems. [ABSTRACT FROM AUTHOR]

Drought stress is one of the major impediments to plant growth. Plant growth-promoting rhizobacteria (PGPR) can mitigate moisture stress in plants by increasing the ability of plant nutrient uptake and transport. In this study, we investigated the root phenotype, mineral nutrients (in leaves, roots, and soil), soil pH, water saturation deficit (WSD), free water content (FWC), and bound water content (BWC) of leaves of two alfalfa varieties, 'Galalxie Max' (drought-tolerant) and 'Saidi 7' (drought-sensitive), in the presence or absence of Bacillus amyloliquefaciens QST713 under drought stress conditions. The results showed that water stress negatively affected both cultivar root morphology (total root length, average diameter, total surface area, and volume) and the contents of K and Fe in leaves, roots, and soil. It also reduced the Mn and Zn contents in the soil while increasing the content of Na in the leaves and soil. Additionally, alfalfa plants under drought stress exhibited higher levels of soil pH, WSD, and BWC but lower contents of FWC and ratios of BWC/FWC in the leaves of both cultivars. However, QST713 application significantly enhanced the total root length, average root diameter, and the contents of K and Fe in alfalfa leaves, roots, and soil, as well as the BWC/FWC ratio in leaves under drought stress conditions. A significant reduction in the Na content was detected in QST713-treated alfalfa leaves and soil under drought stress. Furthermore, QST713 application noticeably decreased soil pH and WSD. The current findings showed that QST713 enhanced the water stress tolerance of alfalfa plants by ameliorating root morphology, reducing soil pH, and improving the BWC/FWC ratio, consequently promoting the accumulation of mineral nutrients (mainly K and Fe). Overall, Bacillus amyloliquefaciens QST713 can serve as a potential green fertilizer in sustainable agriculture to improve soil nutrients and enhance plant production under increasing drought conditions. [ABSTRACT FROM AUTHOR]

Flowering, the transition from the vegetative to the reproductive stage, is vital for reproductive success, affecting forage quality, the yield of aboveground biomass, and seed production in alfalfa. To explore the transcriptomic profile of alfalfa flowering transition, we compared gene expression between shoot apices (SAs) at the vegetative stage and flower buds (FBs) at the reproductive stage by mRNA sequencing. A total of 3,409 DEGs were identified, and based on gene ontology (GO), 42.53% of the most enriched 15 processes were associated with plant reproduction, including growth phase transition and floral organ development. For the former category, 79.1% of DEGs showed higher expression levels in SA than FB, suggesting they were sequentially turned on and off at the two test stages. For the DEGs encoding the components of circadian rhythm, sugar metabolism, phytohormone signaling, and floral organ identity genes, 60.71% showed higher abundance in FB than SA. Among them, MsAP1, an APETALA1 (AP1) homolog of Arabidopsis thaliana, showed high expression in flower buds and co-expressed with genes related to flower organ development. Moreover, ectopic expression of MsAP1 in Arabidopsis resulted in dwarfism and early flowering under long-day conditions. The MsAP1-overexpression plant displayed morphological abnormalities including fused whorls, enlarged pistils, determinate inflorescence, and small pods. In addition, MsAP1 is localized in the nucleus and exhibits significant transcriptional activity. These findings revealed a transcriptional regulation network of alfalfa transition from juvenile phase to flowering and provided genetic evidence of the dual role of MsAP1 in flowering and floral organ development. [ABSTRACT FROM AUTHOR]

Medicago sativa is a high-quality legume forage that is widely cultivated around the world. However, low phosphorus (LP) stress is one of the main limiting factors for its yield and quality. Currently, it is unclear how various alfalfa root types respond to LP stress. Thus, the creeping-rooted M. varia 'Gannong No. 4' (or GN), tap-rooted M. sativa 'Longdong' (or LD), and rhizomatous-rooted M. sativa 'Qingshui' (or QS) were selected to detect changes in growth, metabolites, phytohormones, and organic acids after 34 days of LP stress treatment (0.01 mmol·L−1 KH2PO4). We observed LP stress significantly inhibited plant growth, and 123, 73, and 42 considerably upregulated differential metabolites were categorized into 33, 26, and 20 metabolic pathways in GN, LD, and QS under LP stress, respectively. Amino acids, plant growth regulators, and organic acids are the main metabolites. After 34 d of LP treatment, the plant height, total surface area, leaf length, ground biomass, leaf width, total volume, leaf area, and ZT content of different root types of alfalfa significantly decreased, while the contents of malic acid, citric acid, oxalic acid, IAA, ABA, and GA3 significantly increased. The plant height, leaf length, leaf width, leaf area, total surface area, total volume, ground biomass, root biomass, the number of different metabolites, and the contents of malic, citric, and oxalic acid, and IAA of GN were significantly higher than those of QS and LD under LP stress. From this point of view, GN was more tolerant than LD and QS under the LP condition. [ABSTRACT FROM AUTHOR]

Field dodder (Cuscuta campestris yuncker) is an important parasitic weed that has negative impacts on the growth and yield of alfalfa (Medicago sativa L.). Effective control of field dodder in alfalfa crops is necessary to reduce yield losses. Field studies were conducted to evaluate the efficacy of various herbicides on field dodder in a new-seeded alfalfa crop at three locations (Karaj, Jiroft, and Urmia) in Iran during 2020–2021. The herbicides evaluated were imazethapyr (100 g a.i. ha−1), glyphosate (six doses ranging from 143 to 348 g a.i. ha−1 + ammonium sulfate at 8 kg ha−1), and propyzamide (1250, 1500, and 1750 g a.i. ha−1). The results demonstrated that propyzamide at 1750 g a.i. ha−1 provided complete control of field dodder in terms of percentage change in biomass at first (100%) and second (99%) cuttings of alfalfa in Urmia. The imazethapyr at 100 g a.i. ha−1 provided 96 and 87% control at the first and second cuttings, respectively, whereas propyzamide at 1750 g a.i. ha−1 showed the best efficacy in increasing alfalfa biomass at the first (up to 82%) and the second cuttings (up to 81%), depending on the location. Glyphosate provided reasonable control of the field dodder at the first (73%) and the second cutting (82%). However, at higher rates (307 and 348 g a.i. ha−1), it caused significant reductions in alfalfa biomass due to crop injury, especially at the first cutting. The use of propyzamide at 1750 g a.i. ha−1 is recommended for highly effective control of field dodder in alfalfa crops without compromising the crop yield. [ABSTRACT FROM AUTHOR]

To assess the biocontrol potential of Nigrospora oryzae against Solanum rostratum, the effects of different medium components and fermentation conditions on the biomass of N. oryzae were investigated to determine the optimal medium composition and fermentation conditions. Subsequently, the pathogenicity of S. rostratum increased after artificial inoculation of S. rostratum with N. oryzae. Additionally, the safety of N. oryzae fermentation on seven crops were evaluated. The results revealed that the optimal shake flask culture ratio for N. oryzae was lactose:glycerol:yeast extract:(NH4)2SO4 = 2:2:1:2. The optimal fermentation conditions were as follows: 15 mL of inoculum, 75 mL of fermentation liquid in a 150 mL shake flask, an initial pH of 5.0, a culture temperature of 20 °C, and 4 days of culture. The disease index of S. rostratum after spraying with N. oryzae fermentation agent was 60.00, indicating strong pathogenicity to S. rostratum. Safety evaluation revealed that N. oryzae exhibited no symptoms on Zea mays, S. melongena, Festuca arundinacea, Bromus inermis, or Medicago sativa but showed moderate susceptibility to Triticum aestivum and S. lycopersicum. This study represents the first exploration of the biocontrol potential of N. oryzae against S. rostratum. These findings suggest the potential of N. oryzae for development as a microbial herbicide targeting S. rostratum. [ABSTRACT FROM AUTHOR]

Silage preservation is critical for livestock's stable forage supply during cold seasons and fostering the recycling of agricultural byproducts. We assessed the effects of adding previously fermented juices (PFJ) and different proportions of Sorghum Sudanense (Piper) Stapf (Sudan grass) on the silage quality of Medicago sativa L. (alfalfa). A 50% alfalfa and 50% Sudan grass (M50) mixture exhibited superior performance in sensory evaluation and fermentation quality. The addition of PFJ (PFJI group) further enhanced quality, and increasing the proportion of alfalfa improved the nutritional quality. However, PFJI had no significant effect on the degradation rates of these nutritional indicators (p > 0.05). As alfalfa proportion increased, the disappearance rates of dry matter, crude protein, neutral detergent fiber, and acid detergent fiber in silage increased. Principal component, correlation, and membership function analyses revealed that treatment with M50 without PFJ (PFJ0+M50) had the best effect, followed by treatment with 25% alfalfa and 75% Sudan grass (PFJ0+M25) and 100% alfalfa (PFJ0+M0). Therefore, the mixing ratio of alfalfa and Sudan grass should be maintained between 25% and 50% to optimize the nutritional and fermentation qualities of silage. These findings offer crucial guidance for alfalfa silage preparation, promoting enhanced livestock industry productivity and sustainable agricultural development. [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]

In this study, the wear characteristics of the die were tested and analyzed through compaction tests, and the distribution of wear depth along the direction toward the extrusion outlet was obtained. A discrete element method (DEM) model of the die's wear process was established. The results show that the severe wear area is located near the stop position of the compression rod, forming a plow-shaped wear area along the extrusion direction, accompanied by fatigue peeling. The wear depth gradually decreases towards the extrusion outlet. The DEM model partially reveals the occurrence of the wear phenomenon, but the particle motion speed deviates from the actual situation. The maximum compression force value range during the DEM compression stage is within the actual maximum compression force value range, and the relative error range of the average maximum compression force is less than 2%. By verifying the formula to calibrate the model, the calibrated model is compared with the actual mold wear, and the predicted value is close to the actual test result. The DEM can be used to explore the wear mechanism and predict the die's wear failure process, laying the foundation for optimizing die wear resistance design. [ABSTRACT FROM AUTHOR]

Pollutant degradation and heavy-metal resistance may be important features of the rhizobia, making them promising agents for environment cleanup biotechnology. The degradation of phenanthrene, a three-ring polycyclic aromatic hydrocarbon (PAH), by the rhizobial strain Rsf11 isolated from the oil-polluted rhizosphere of alfalfa and the influence of nickel ions on this process were studied. On the basis of whole-genome and polyphasic taxonomy, the bacterium Rsf11 represent a novel species of the genus Neorhizobium, so the name Neorhizobium phenanthreniclasticum sp. nov. was proposed. Analysis of phenanthrene degradation by the Rsf1 strain revealed 1-hydroxy-2-naphthoic acid as the key intermediate and the activity of two enzymes apparently involved in PAH degradation. It was also shown that the nickel resistance of Rsf11 was connected with the extracellular adsorption of metal by EPS. The joint presence of phenanthrene and nickel in the medium reduced the degradation of PAH by the microorganism, apparently due to the inhibition of microbial growth but not due to the inhibition of the activity of the PAH degradation enzymes. Genes potentially involved in PAH catabolism and nickel resistance were discovered in the microorganism studied. N. phenanthreniclasticum strain Rsf11 can be considered as a promising candidate for use in the bioremediation of mixed PAH–heavy-metal contamination. [ABSTRACT FROM AUTHOR]

Water is an important constraint on alfalfa (Medicago sativa) production in arid and semiarid areas, and alternate irrigation in root areas has water-saving potential for alfalfa production. To investigate the impact of alternate partial root-zone irrigation (APRI) on the rhizosphere soil microorganisms of alfalfa, this study subjected alfalfa plants to different irrigation methods and irrigation levels. The growth status and rhizosphere soil microbial community diversity of alfalfa plants under alternate root-zone watering treatment were analyzed through laboratory experiments and high-throughput sequencing. The results showed that at soil moisture levels of 80% field moisture capacity (FMC) and 60% FMC, APRI had no significant impact on the biomass or nodule number of alfalfa. However, 40% FMC significantly reduced the individual plant dry weight, chlorophyll content, and nodule number of the alfalfa plants. APRI increased the relative abundance of Actinomycetes in the alfalfa rhizosphere soil. Moreover, at 60% FMC, the MBC and MBN of rhizosphere, relative abundance of Actinobacteria and unclassified K fungi and Chao 1 index of bacteria significantly increased under APRI treatment. While relative abundance of Ascomycetes and Proteobacteria in the alfalfa rhizosphere significantly reduced under 60% FMC + APRI treatment. In summary, under the same irrigation conditions, APRI did not significantly affect the growth of alfalfa in the short term. And 60%FMC + APRI treatment did significantly affect the groups, structure and diversity of the rhizosphere soil microbial communities. [ABSTRACT FROM AUTHOR]

Introduction: Lucerne (Medicago sativa), is a cornerstone of China's livestock industry, however, due to the backward agronomic strategies and technology, lucerne in China faces cultivation challenges that result in lower productivity and quality than global standards. Therefore, we undertook a meta-analysis to evaluate the impact of five distinct fertilization types on lucerne yield and nutritional quality in various locations in China. The fertilizer practices included manure application, combined mineral fertilizer and manure application (FM), biological fertilizer application, unbalanced application of two or more mineral fertilizer types, and balanced mineral fertilizer application. Furthermore, we investigate influential factors of yield and quality of lucerne under fertilization, including climatic variables (mean annual precipitation, mean annual temperature), initial soil properties (soil organic carbon; total nitrogen, pH), and agronomic factors (seeding rate, harvest frequency, and lucerne stand age). Methods: Our study analyzed 53 published papers to discern the most beneficial fertilizer for enhancing lucerne yield and nutritional quality. Results and discussion: The results showed that the fertilizer practices, on average, significantly improved yield by 31.72% and crude protein content by 11.29%, with FM emerging as the most effective, this is because mineral fertilizers provide available nutrients for lucerne, manure provides essential organic matter for microorganisms and improve soil properties. In addition, the fertilizer practices significantly reduced neutral and acid detergent fiber contents by 6.28% and 8.50%, respectively, while increasing ash content and relative feeding value. Furthermore, climatic variables, soil properties, and planting system factors such as sowing date and harvest frequency significantly affected yield and nutritional quality. The practical implications of our results emphasize the need for balanced and strategic fertilizer application to optimize lucerne production and highlight the potential to adjust cultivation practices according to environmental conditions. Balanced and strategic fertilizer application can simultaneously improve soil properties, enhance soil carbon sequestration, and reduce the emission of greenhouse gases from the soil, which is a vital measure for realizing sustainable agricultural development. [ABSTRACT FROM AUTHOR]

This article follows-up on our recently published work, which evaluated the impact of the addition of an alfalfa leaf-derived adsorbent in the aflatoxin B1 (AFB1)-contaminated diet in regard to the production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology of turkey poults. This paper presents complementary results on microbial community, ileal morphology, barrier function, and immunity. For this purpose, 350 1-day-old female turkey poults were randomly distributed into five groups: (1) Control, AFB1-free diet; (2) AF, AFB1-contaminated diet at 250 ng/g; (3) alfalfa, AFB1-free diet + 0.5% (w/w) adsorbent; (4) alfalfa + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) adsorbent; and (5) YCW + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) commercial yeast cell wall-based adsorbent (reference group). In general, in the AF group, the growth of opportunistic pathogens was promoted, which lead to gut dysbacteriosis, mainly influenced by Streptococcus lutetiensis. Conversely, a significant increase in beneficial bacteria (Faecalibacterium and Coprococcus catus) was promoted by the addition of the plant-based adsorbent. Moreover, the AF group had the lowest villus height and a compromised barrier function, as evidenced by a significant (p < 0.05) increase in fluorescein isothiocyanate dextran (FITC-d), but these negative effects were almost reversed by the addition of the alfalfa adsorbent. Furthermore, the AF + YCW and alfalfa + AF groups exhibited a significant increase in the cutaneous basophil hypersensitivity response compared to the rest of the experimental groups. Taken together, these results pointed out that the alfalfa counteracts the adverse effects of AFB1 in poults, facilitating the colonization of beneficial bacteria and improving the barrier function of the turkey poults. [ABSTRACT FROM AUTHOR]

To enhance the selenium (Se) intake of the general public, the present study implemented biofortification techniques in alfalfa sprouts. Alfalfa sprouts possess unique nutritional value and provide an optimal Se-enriched supplemental Se source. The impact of sodium selenite (Na2SeO3) on alfalfa shoot germination, shoot length, and biomass was assessed experimentally, and changes in the antioxidant capacity of sprouts treated with optimal Se concentrations were investigated. In addition, the transcriptome of alfalfa sprouts treated with the optimal Na2SeO3 concentration was sequenced. Gene co-expression networks, constructed through differential gene analysis and weighted gene co-expression network analysis, were used to identify the core genes responsible for Se enrichment in alfalfa sprouts. The findings of the present study offer novel insights into the effects of Se treatment on the nutrient composition of alfalfa sprouts, in addition to introducing novel methods and references that could facilitate production of Se-enriched alfalfa sprouts and associated products. [ABSTRACT FROM AUTHOR]

The crude protein (CP) content is an important determining factor for the quality of alfalfa, and its accurate and rapid evaluation is a challenge for the industry. A model was developed by combining Fourier transform infrared spectroscopy (FTIS) and chemometric analysis. Fourier spectra were collected in the range of 4000~400 cm−1. Adaptive iteratively reweighted penalized least squares (airPLS) and Savitzky–Golay (SG) were used for preprocessing the spectral data; competitive adaptive reweighted sampling (CARS) and the characteristic peaks of CP functional groups and moieties were used for feature selection; partial least squares regression (PLSR) and random forest regression (RFR) were used for quantitative prediction modelling. By comparing the combined prediction results of CP content, the predictive performance of airPLST-cars-PLSR-CV was the best, with an R P 2 of 0.99 and an RMSEP of 0.053, which is suitable for establishing a small-sample prediction model. The research results show that the combination of the PLSR model can achieve an accurate prediction of the crude protein content of alfalfa forage, which can provide a reliable and effective new detection method for the crude protein content of alfalfa forage. [ABSTRACT FROM AUTHOR]

Background: Alfalfa (Medicago sativa L.) experiences many negative effects under salinity stress, which may be mediated by recurrent selection. Salt-tolerant alfalfa may display unique adaptations in association with rhizobium under salt stress. Results: To elucidate inoculation effects on salt-tolerant alfalfa under salt stress, this study leveraged a salt-tolerant alfalfa population selected through two cycles of recurrent selection under high salt stress. After experiencing 120-day salt stress, mRNA was extracted from 8 random genotypes either grown in 0 or 8 dS/m salt stress with or without inoculation by Ensifer meliloti. Results showed 320 and 176 differentially expressed genes (DEGs) modulated in response to salinity stress or inoculation x salinity stress, respectively. Notable results in plants under 8 dS/m stress included upregulation of a key gene involved in the Target of Rapamycin (TOR) signaling pathway with a concomitant decrease in expression of the SNrK pathway. Inoculation of salt-stressed plants stimulated increased transcription of a sulfate-uptake gene as well as upregulation of the Lysine-27-trimethyltransferase (EZH2), Histone 3 (H3), and argonaute (AGO, a component of miRISC silencing complexes) genes related to epigenetic and post-transcriptional gene control. Conclusions: Salt-tolerant alfalfa may benefit from improved activity of TOR and decreased activity of SNrK1 in salt stress, while inoculation by rhizobiumstimulates production of sulfate uptake- and other unique genes. [ABSTRACT FROM AUTHOR]

Objective: A study was conducted to quantify the performance differences of the near-infrared spectroscopy (NIRS) calibration models developed with different degrees of hay sample preparations. Methods: A total of 227 imported alfalfa (Medicago sativa L.) and another 360 imported timothy (Phleum pratense L.) hay samples were used to develop calibration models for nutrient value parameters such as moisture, neutral detergent fiber, acid detergent fiber, crude protein, and in vitro dry matter digestibility. Spectral data of hay samples prepared by milling into 1-mm particle size or unground were separately regressed against the wet chemistry results of the abovementioned parameters. Results: The performance of the developed NIRS calibration models was evaluated based on R², standard error, and ratio percentage deviation (RPD). The models developed with ground hay were more robust and accurate than those with unground hay based on calibration model performance indexes such as R² (coefficient of determination), standard error, and RPD. Although the R² of calibration models was mainly greater than 0.90 across the feed value indexes, the R² of cross-validations was much lower. The R² of cross-validation varies depending on feed value indexes, which ranged from 0.61 to 0.81 in alfalfa, and from 0.62 to 0.95 in timothy. Estimation of feed values in imported hay can be achievable by the calibrated NIRS. However, the NIRS calibration models must be improved by including a broader range of imported hay samples in the modeling. Conclusion: Although the analysis accuracy of NIRS was substantially higher when calibration models were developed with ground samples, less sample preparation will be more advantageous for achieving rapid delivery of hay sample analysis results. Therefore, further research warrants investigating the level of sample preparations compromising analysis accuracy by NIRS. [ABSTRACT FROM AUTHOR]

Prolonged drought exacerbated by climate change in the Mesilla Valley, one of the major agricultural areas of New Mexico, USA, is causing a shortage of surface water from the Rio Grande for irrigation. Farmers in the Valley are using groundwater for irrigation and complementing it with limited surface water from the river (Rio Grande). Managing irrigation water better is vital to sustaining agriculture in the Valley. Remote sensing (RS)-based crop evapotranspiration (ETa) models offer significant advantages over traditional methods. The ET maps generated by these RS models provide valuable information that can be used to manage irrigation water and crops in water-scarce areas. This study used METRIC and SSEBop RS models to map the ET of alfalfa on a private farm that is managed as commonly practiced in the Valley. The integrated ET values of the two models are compared to those of the ETa measured using the eddy covariance method. The comparison showed that 91.55% of the variability in SSEBop ETa estimates can be explained by the variability in the METRIC ETa estimates, and the variability in eddy covariance ETa can explain 93.07% of the variability in METRIC ETa and 86.01% in the SSEBop Eta estimates. Both METRIC and SSEBop reflected the ETa of alfalfa during full growth and harvesting periods. However, the absolute percent mean relative difference (MRD) of ET was higher for two out of three cuttings by SSEBop (>32%) compared to those for METRIC and eddy covariance. The spatiotemporal variabilities in crop ET estimates using METRIC and SSEBop showed a need to improve on-farm irrigation conveyance and on-the-field irrigation efficiency. Overall, RS models can provide spatiotemporal maps of ET that can be used for decision-making to manage irrigation water better and improve crop yield on a field, farm, and regional scale. [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]

In the field of agro-grassland, the synergism of phosphate (P) fertilization and Trichoderma inoculation in alfalfa production and the underlying mechanism of rare and abundant microbes that regulate rhizosphere soil processes in various environments are key but rarely studied topics. Here, we conducted field research through the inoculation/noninoculation of Trichoderma and five levels of phosphorus fertilizer to explore the biological relationships of rhizosphere soil properties, rare and abundant taxa, and alfalfa yields. Our results demonstrate that using Trichoderma inoculation and 15 g/m2 of phosphorus fertilizer exhibited optimal alfalfa yield compared with other treatments. alfalfa yields significantly (R2 = 0.33; p < 0.001) increased along with soil fertility. Both rare (RANOSIM = 0.900; p = 0.001) and abundant (RANOSIM = 0.769; p = 0.001) bacterial communities were significantly different under Trichoderma inoculation and P fertilization in a nonmetric multidimensional scaling (NMDS) analysis. Furthermore, different ecological processes dominated the rare and abundant bacterial community assembly. PLS-PM analysis showed that Trichoderma inoculation positively regulated the abundant bacteria community and P fertilization regimes manipulated the rare bacteria community, synergistically contributing to alfalfa yields. Overall, this article believes that inoculation with Trichoderma and appropriate application of phosphorus fertilizer can significantly increase alfalfa yield and affect soil enzyme activity, and the rhizosphere soil abundant and rare bacterial community characteristics have different responses to the synergistic effect of Trichoderma and phosphorus fertilizer. Our research emphasizes the fundamental role of abundant and rare microbes in maintaining crop production using Trichoderma inoculation and P fertilization. Therefore, distinguishing rare and abundant species is beneficial to comprehensively understanding microbial-driven processes and providing theoretical support for maintaining ecosystem productivity. [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]

The long-term storage of coal gangue (CG) mountains causes serious environmental problems such as water and air pollution. Thus, sustainable reclamation practices are urgently needed to minimize the environmental impacts brought by CG mountains. Pikovskaya medium was employed to screen microorganisms, which were subsequently utilized to promote the solubilization of CG. XRF, SEM, XRD, and HPLC techniques were employed to characterize the CG before and after bacterial treatment. In this study, we have successfully isolated and purified a bacterial strain, identified as Stenotrophomonas bentonitica BII-R7, which possesses the ability to facilitate the solubilization of nutrient elements from CG. Factors including initial inoculation ratio, incubation time, CG particle size, CG concentration, pH, and temperature were examined to investigate their effects on the biosolubilization of CG. Furthermore, the mechanism underlying the CG solubilization was also probed. Our data demonstrated that low-molecular-weight organic acids, such as acetic acid and formic acid, may harbor a crucial role in promoting the solubilization of CG. Lastly, we found that Stenotrophomonas bentonitica BII-R7, in conjugation with CG, can increase the alfalfa seed germination percentage and promote the growth of alfalfa. Together, these data provide evidence that bacterial-treated CG can be utilized for soil improvement and land reclamation. [ABSTRACT FROM AUTHOR]

Low temperature is the most common abiotic factor that usually occurs during the seed germination of alfalfa (Medicago sativa L.). However, the potential regulatory mechanisms involved in alfalfa seed germination under low temperature stress are still ambiguous. Therefore, to determine the relevant key genes and pathways, the phenotypic and transcriptomic analyses of low-temperature sensitive (Instict) and low-temperature tolerant (Sardi10) alfalfa were conducted at 6 and 15 h of seed germination under normal (20 °C) and low (10 °C) temperature conditions. Germination phenotypic results showed that Sardi10 had the strongest germination ability under low temperatures, which was manifested by the higher germination-related indicators. Further transcriptome analysis indicated that differentially expressed genes were mainly enriched in galactose metabolism and carbon metabolism pathways, which were the most commonly enriched in two alfalfa genotypes. Additionally, fatty acid metabolism and glutathione metabolism pathways were preferably enriched in Sardi10 alfalfa. The Weighted Gene Co-Expression Network Analysis (WGCNA) suggested that genes were closely related to galactose metabolism, fatty acid metabolism, and glutathione metabolism in Sardi10 alfalfa at the module with the highest correlation (6 h of germination under low temperature). Finally, qRT-PCR analysis further validated the related genes involved in the above pathways, which might play crucial roles in regulating seed germination of alfalfa under low temperature conditions. These findings provide new insights into the molecular mechanisms of seed germination underlying the low temperature stress in alfalfa. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) is a vitally important perennial fodder legume worldwide. Given their particular traits, alfalfa crop wild relatives (CWRs) could be used to develop cultivars that can tolerate extreme environmental and climatic conditions. Until now, researchers have overlooked the composition and structure of bacterial communities in the root zone of alfalfa and its relevant CWRs and their influence on forage performance under actual field conditions. In this study, high-throughput sequencing of 16S rRNA analysis was performed to investigate the diversity and assemblies of bacterial communities in the bulk soil and in the root zone of individual field-grown Medicago plants arranged in a honeycomb selection design. The plants used in this study were M. sativa × M. arborea hybrids (Genotypes 6 and 8), the closely-related M. sativa nothosubsp. varia (Martyn) Arcang. (Genotype 13), and M. sativa ssp. sativa (Genotype 20). The bacterial communities in the root-zone samples and the assemblies in the bulk soil differed significantly. Genotype 13 was found to have distinct bacterial assemblies from the other genotypes while exhibiting the lowest forage productivity. These findings suggest that plant productivity may influence the composition of bacterial communities in the root zone. Biomarker analysis conducted using linear discriminant analysis (LDA) revealed that only members of the Rhizobiales order were enriched in the M. sativa nothosubsp. varia root zone whereas taxa belonging to Sphingomonas and various Bacteriodota were enriched in the other genotypes. Of the shared taxa identified in the root zone of the Medicago lines, the abundance of specific taxa, namely, Flavisolibacter, Stenotrophomonas, and Sphingomonas, were positively associated with forage yield. This pioneering study, in which the root zones of individual Medicago plants under actual field conditions were examined, offers evidence of differences in the bacterial composition of alfalfa genotypes with varying genetic backgrounds. Its findings indicate that particular bacterial taxa may favorably influence plant performance. This study covered the first six months of crop establishment and paves the way for further investigations to advance understanding of how shifts in bacterial assemblies in alfalfa roots affect plant performance over time. [ABSTRACT FROM AUTHOR]

Anthracnose caused by various species of Colletotrichum is one of the most prevalent diseases in alfalfa worldwide that not only reduces forage yields but also severely compromises forage quality. A comprehensive survey was conducted in 2020 in the main production regions of northern China. The survey results showed that alfalfa anthracnose is prevalent in northern China, with the disease incidence ranging from 9% to 45% and the disease index from 5 to 17 (maximum possible score: 100). In total, 24 isolates were collected and identified as three Colletotrichum species (C. trifolii, C. truncatum and C. americae-borealis) based on morphological characteristics and phylogenetic analysis (combined sequences ITS, HIS3, ACT and GAPDH). The three species displayed remarkable environmental adaptability, exhibiting a capacity for growth, sporulation and conidial germination in temperatures ranging from 4 to 35 °C and in different nutrient conditions. Pathogenicity assays showed that C. trifolii was more virulent than the other two species, although the growth vigor (in terms of colony diameter, sporulation and conidial germination) of C. truncatum was the greatest. [ABSTRACT FROM AUTHOR]

High-voltage discharge plasma drying (HVDPD) is a non-thermal and efficient drying technique. In this study, we compared the drying characteristics and physicochemical properties of alfalfa dried via HVDPD, hot-air drying (HAD), and natural air-drying (AD) and analyzed the experimental results using infrared spectroscopy, scanning electron microscopy, colorimetry, and other detection techniques. The results showed that HVDPD had a higher drying rate than AD and saved more energy than HAD. Alfalfa dried via HVDPD had the most intact internal structure, the highest rehydration ratio (RR), the best color quality, and the best nutrient retention, resulting in the highest relative quality and feeding value. Therefore, HVDPD is an excellent drying technique for alfalfa, providing a new and effective way to improve the storage, transportation, and drying quality of alfalfa. [ABSTRACT FROM AUTHOR]

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

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.

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.

Abstract Soil salinization poses a serious threat to crop growth. The selection of appropriate soil modifiers and water-soluble fertilizers for saline soils represents a crucial method for enhancing crop yields. The modifiers and medium-element water-soluble fertilizers were prepared using hydrochar derived from rice straw. Two distinct experiments were designed to study the effect of modifiers and water-soluble fertilizers on saline soils. The first experiment, designated as the “Soil Cultivation Experiment” , sought to investigate the impact of various modifiers on soil quality. The second experiment, designated as the “Method of Field Micro-Area Experiment”, aimed to assess the influence of water-soluble fertilizers on saline soils. The results showed that the application of modifiers and water-soluble fertilizers significantly enhanced comprehensive soil physical and chemical properties, crop growth, soil enzyme activity, and other key indicators in saline and alkaline soils. The optimal dosage of the modifier was 20 g/kg, which reduced the pH value from 8.62 to 8.21 and the decreased alkalinity by 8.26%. Furthermore, their application effectively boosted nutrient levels, including organic matter, and increased soil enzyme activity. The biomass of alfalfa showed enhancements of 63.01% and 20.87% and the biomass of leymus chinensis increased by 29.39% and 9.02% for the two batches, respectively. Notably, the application of water-soluble fertilizer yielded achieved superior results. This study also provided a theoretical basis for their future application in soda saline–alkali soil.

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]

Under the pressures of global climate change and human activities, the carrying capacity of water and soil resources in agricultural lands has decreased, and the traditional models of agricultural development are no longer sustainable. Land degradation, groundwater quality reduction and ecosystem instability are the consequences of agricultural development without considering sustainability indicators. This article aims to investigate the use of variable relationship analysis and Bayesian network methods to analyze and investigate the relationship between irrigation in agriculture and the sustainability of the groundwater ecosystem. Descriptive statistics of agriculture including cultivation pattern, time, precipitation, irrigation, and land slope were analyzed and combined with the simulated characteristics of groundwater including specific yield, hydraulic conductivity and hydrodynamic diffusion coefficients. Five crops of wheat, barley, paddy, alfalfa, and potato were studied to evaluate the effect of plants on the pattern of nitrate release due to irrigation and fertilization in agriculture. The results showed that managing the amount of fertilizer and the volume of irrigation can positively affect the nitrate distribution pattern in the groundwater even in a short period of time. HIGHLIGHTS The relationships between variables are used to build a logical model for analysis.; The quantity and quality of irrigation water is incorporated into the system for estimating the nitrate concentration in groundwater.;

The effect of different irrigation methods (sprinkler, surface, surface-drip, and subsurface-drip) using treated wastewater, on alfalfa yield quantity and quality, was studied under semi-arid conditions. Randomized complete block design considering irrigation methods on 5 × 5 m plots replicated four times. Applied irrigation water was based on the Penman–Monteith equation using the FAO Cropwat software and accounting for the efficiencies of used irrigation systems. Surface irrigation gave the highest alfalfa fresh yield without significant difference compared to subsurface-drip. Average fresh production was 123, 120, 109, and 91 tons/h for surface, subsurface-drip, surface-drip, and sprinkler irrigation, respectively. Alfalfa fresh weight from subsurface-drip irrigation was 32 and 10% higher compared to sprinkler and surface-drip irrigation. Alfalfa dry weight from subsurface-drip irrigation was 10, 21, and 47% higher compared to surface-drip, surface, and sprinkler irrigation, respectively. N percentage in alfalfa leaves was significantly lower by 12% under subsurface-drip irrigation as compared to the other irrigation methods. Escherichia coli (E. coli) and fecal coliform (FC) were not detected on alfalfa leaves using subsurface-drip irrigation. E. coli and FC counts were high on leaves using sprinkler and surface irrigation. Subsurface-drip irrigation may be adapted as an efficient irrigation method when using non-conventional water under semi-arid conditions. HIGHLIGHTS Irrigation study: Examined methods using treated wastewater in semi-arid areas.; Subsurface-drip: Tops in dry yield, matches in fresh yield.; Health benefits: Subsurface-drip reduces E. coli and fecal coliform on alfalfa.; Smart water use: Advocates calculated use with FAO Cropwat insights.; Alternative water: Recommends subsurface-drip for treated wastewater in semi-arid regions.;