Impact of Partial Replacement of Clover by Anise, Basil, Mint and Fennel Remnants in Rabbits Diet on Blood Constituents.

<b>Background and Objective:</b> The clover plant (alfalfa) is considered the primary fiber source in the rabbit diet in Mediterranean Sea countries like Egypt, so researchers are always trying to find alternatives and aromatic and medicinal plant remnants could be one of them. So, this study was designed to determine the effects of some aromatic and medicinal plant remnants on New Zealand white rabbits' blood hematological and biochemical aspects. <b>Materials and Methods:</b> A total of 108 weaned white New Zealand rabbits at five weeks of age were used to consider the effect of using remnants of mint, fennel, basil and anise with or without probiotics to replace 50% from alfalfa hay in rabbits' diets. Four remnants were obtained after etheric oil distillation and were incorporated in rabbit diets at level 17.5% without probiotics and with probiotics (replacement 50% of alfalfa hay). Rabbits were randomly assigned into nine experimental groups; the experimental period lasted eight weeks. Rabbit blood hematological and blood biochemical were analyzed. <b>Results:</b> The highest values of RBC 6.03 µL, HCT 37.13%, WBC 12.70 µL and lymph percentage were found in the basil+probiotics group. In contrast, the highest value of hemoglobin (HGB 10,50 g/dL), MCV 64.13 fl, MCH 23.27pg, MCHC 36.40 g/dL, PLT 463 µL, urea 50.33 mg/dL and creatinine 1.30 mg/dL were found in anise+probiotic group. In contrast, RDW-CV 33.17%, Mid 13.17 µL, granulocytes (Gran 7.13 µL) and PDW 16.73 in the mint group. Furthermore, RDW-SD (34.40 fl) and procalcitonin (PCT 0.35%) were found in the control group and the highest values ALT 142 IU/L and AST 77.33 IU/L were found in the fennel group. The highest albumin value (3.10 g/dL) was found in the anise group and the highest total protein (TP 5.23 g/dL) was found in the mint+probiotic group. <b>Conclusion:</b> The results proved that using these medicinal plant remnants and probiotics as substitutes for half the amount of alfalfa used in the diet of New Zealand white rabbits did not have a negative effect and improved their health condition.

Forage Height and Above-Ground Biomass Estimation by Comparing UAV-Based Multispectral and RGB Imagery.

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

Screening of mtr-miR156a from exosomes of dairy cow blood to milk and its regulatory effect on milk protein synthesis in BMECs.

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

Accumulation of microcystin toxin in irrigation water and alfalfa (Medicago sativa) forage plant, and assessing the potential risk to animal health.

Microcystin (MC) toxin produced by cyanobacteria has become a significant concern for societies worldwide. The risk of MC in drinking water has been assessed to human health. Nonetheless, its risk to animal health has not been thoroughly evaluated. This study investigated MCs in irrigation water and alfalfa plant from nearby farmlands. Both irrigation water and alfalfa shoots contained greater MC concentrations (1.8-17.4 µg L-1 and 0.053-0.128 µg g-1) during summer than winter (2.4 µg L-1 and 0.017 µg g-1). These MC concentrations showed a correlation with the predominance of cyanobacteria in the sites, triggering the potential risk of these microorganisms in irrigation waters. Accordingly, there would be a high risk (risk quotient, RQ > 1) during summer and a moderate risk (0.1

Phytoremediation of molybdenum (Mo)-contaminated soil using plant and humic substance.

The severity of soil molybdenum (Mo) pollution is increasing, and effective management of contaminated soil is essential for the sustainable development of soil. To investigate this, a pot experiment was carried out to assess the impact of different rates of humic acid (HA) and fulvic acid (FA) on the mobility of Mo in soil solution and its uptake by alfalfa, wheat and green bristlegrass. The concentration of Mo in Plants and soil was determined using an Atomic Absorption Spectrophotometer. The findings revealed that the application of HA led to an increase in Mo accumulation in the shoot and root of green bristlegrass and wheat, ranging from 10.56 % to 28.73 % and 62.15-115.79 % (shoot), and 17.52-46.53 % and 6.29-81.25 % (root), respectively. Nonetheless, the use of HA resulted in a slight inhibition of plant Mo uptake, leading to reduced Mo accumulation in alfalfa roots compared to the control treatment (from 3284.49 mg/kg to 2140.78-2813.54 mg/kg). On the other hand, the application of FA decreased Mo accumulation in the wheat shoot (from 909.92 mg/kg to 338.54-837.45 mg/kg). Furthermore, the bioavailability of green bristlegrass (with HA) and wheat (with FA) decreased, and the percentage of residual fraction of Mo increased (from 0.39 % to 0.78-0.96 %, from 3.95 % to 3.97∼ 4.34 %). This study aims to elucidate the ternary interaction among Mo, humic substances, and plants (alfalfa, wheat, and green bristlegrass), to enhance both the activation and hyperaccumulation of Mo simultaneously.

Satellite observations indicate that chia uses less water than other crops in warm climates.

Many parts of the world face severe and prolonged drought conditions, stressing the sustainability of water resources and agriculture. Transitioning to water-efficient crops is one strategy that can help adapt to water scarcity. An emerging drought-tolerant crop of interest is chia (Salvia hispanica). Yet, no study has compared its large-scale water use dynamics to those of widely established crops across the globe. Here, we use satellite data over multiple years to assess the water use efficiency of chia, alfalfa, corn, and soybean globally. Results show that chia consumed 13-38% less water than alfalfa, corn, and soy and assimilated 14-20% more carbon per amount of water used. Substituting 10% of Southwest United States alfalfa cultivation with chia would save 184 million liters of water per growing season, equivalent to the annual water consumption of 1,300 households. Future research shall explore the economic, societal, and environmental ramifications of substituting alfalfa with chia in dry areas worldwide. These insights can guide decision-makers in promoting sustainable agriculture and water resource management.

Control of inorganic and organic phosphorus molecules on microbial activity, and the stoichiometry of nutrient cycling in soils in an arid, agricultural ecosystem.

Background: The dynamics of carbon (C), nitrogen (N), and phosphorus (P) in soils determine their fertility and crop growth in agroecosystems. These dynamics depend on microbial metabolism, which in turn depends on nutrient availability. Farmers typically apply either mineral or organic fertilizers to increase the availability of nutrients in soils. Phosphorus, which usually limits plant growth, is one of the most applied nutrients. Our knowledge is limited regarding how different forms of P impact the ability of microbes in soils to produce the enzymes required to release nutrients, such as C, N and P from different substrates. Methods: In this study, we used the arable layer of a calcareous soil obtained from an alfalfa cropland in Cuatro Cienegas, México, to perform an incubation experiment, where five different phosphate molecules were added as treatments substrates: three organic molecules (RNA, adenine monophosphate (AMP) and phytate) and two inorganic molecules (calcium phosphate and ammonium phosphate). Controls did not receive added phosphorus. We measured nutrient dynamics and soil microbial activity after 19 days of incubation. Results: Different P molecules affected potential microbial C mineralization (CO2-C) and enzyme activities, specifically in the organic treatments. P remained immobilized in the microbial biomass (Pmic) regardless of the source of P, suggesting that soil microorganisms were limited by phosphorus. Higher mineralization rates in soil amended with organic P compounds depleted dissolved organic carbon and increased nitrification. The C:N:P stoichiometry of the microbial biomass implied a change in the microbial community which affected the carbon use efficiency (CUE), threshold elemental ratio (TER), and homeostasis. Conclusion: Different organic and inorganic sources of P affect soil microbial community structure and metabolism. This modifies the dynamics of soil C, N and P. These results highlight the importance of considering the composition of organic matter and phosphate compounds used in agriculture since their impact on the microbial activity of the soil can also affect plant productivity.

Biocompatible Nano-Cocrystal Engineering for Targeted Herbicide Delivery: Enhancing Efficacy through Stimuli-Responsive Release and Reduced Environmental Losses.

In addressing the critical challenges posed by the misuse and inefficiency of traditional pesticides, we introduce a Nano-Cocrystal material composed of the herbicide clopyralid and coformer phenazine. Developed through synergistic supramolecular self-assembly and mechanochemical nanotechnology, this Nano-Cocrystal significantly enhances pesticide performance. It exhibits a marked improvement in stability, with reductions in hygroscopicity and volatility by approximately 38%. Moreover, it intelligently modulates release according to environmental factors, such as temperature, pH, and soil inorganic salts, demonstrating decreased solubility by up to four times and improved wettability and adhesion on leaf surfaces. Importantly, the herbicidal activity surpasses that of pure clopyralid, increasing suppression rates of Medicago sativa L. and Oxalis corniculata L. by up to 27% at the highest dosage. This Nano-Cocrystal also shows enhanced crop safety and reduced genotoxicity compared to conventional formulations. Offering a blend of simplicity, cost-effectiveness, and robust stability, our findings contribute a sustainable solution to agricultural practices, favoring the safety of nontarget organisms.

Genome-wide identification of the EIN3/EIL transcription factor family and their responses under abiotic stresses in Medicago sativa.

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.

Inversion model of soil salinity in alfalfa covered farmland based on sensitive variable selection and machine learning algorithms.

Purpose: Timely and accurate monitoring of soil salinity content (SSC) is essential for precise irrigation management of large-scale farmland. Uncrewed aerial vehicle (UAV) low-altitude remote sensing with high spatial and temporal resolution provides a scientific and effective technical means for SSC monitoring. Many existing soil salinity inversion models have only been tested by a single variable selection method or machine learning algorithm, and the influence of variable selection method combined with machine learning algorithm on the accuracy of soil salinity inversion remain further studied. Methods: Firstly, based on UAV multispectral remote sensing data, by extracting the spectral reflectance of each sampling point to construct 30 spectral indexes, and using the pearson correlation coefficient (PCC), gray relational analysis (GRA), variable projection importance (VIP), and support vector machine-recursive feature elimination (SVM-RFE) to screen spectral index and realize the selection of sensitive variables. Subsequently, screened and unscreened variables as model input independent variables, constructed 20 soil salinity inversion models based on the support vector machine regression (SVM), back propagation neural network (BPNN), extreme learning machine (ELM), and random forest (RF) machine learning algorithms, the aim is to explore the feasibility of different variable selection methods combined with machine learning algorithms in SSC inversion of crop-covered farmland. To evaluate the performance of the soil salinity inversion model, the determination coefficient (R2), root mean square error (RMSE) and performance deviation ratio (RPD) were used to evaluate the model performance, and determined the best variable selection method and soil salinity inversion model by taking alfalfa covered farmland in arid oasis irrigation areas of China as the research object. Results: The variable selection combined with machine learning algorithm can significantly improve the accuracy of remote sensing inversion of soil salinity. The performance of the models has been improved markedly using the four variable selection methods, and the applicability varied among the four methods, the GRA variable selection method is suitable for SVM, BPNN, and ELM modeling, while the PCC method is suitable for RF modeling. The GRA-SVM is the best soil salinity inversion model in alfalfa cover farmland, with Rv 2 of 0.8888, RMSEv of 0.1780, and RPD of 1.8115 based on the model verification dataset, and the spatial distribution map of soil salinity can truly reflect the degree of soil salinization in the study area. Conclusion: Based on our findings, the variable selection combined with machine learning algorithm is an effective method to improve the accuracy of soil salinity remote sensing inversion, which provides a new approach for timely and accurate acquisition of crops covered farmland soil salinity information.

Investigating the chemical composition, digestion and fermentation of Alternanthera sessilis red plant and its effect on the performance of fattening lambs.

The present experiment was conducted to evaluate the nutritional value of red Alternanthera sessilis for fattening lambs when they were replaced with alfalfa forage. Forty growing lambs with an average weight of 21.12 kg and an age of 5 months were randomly assigned to four experimental treatments. Growth performance, nutrient digestibility, rumen fermentation parameters, protozoa population, blood parameters, and composition of carcass components were evaluated. The results of this experiment showed that the use of Alternanthera sessilis in the diet significantly reduced feed intake, increased the average daily weight gain, and improved the feed conversion ratio compared to the control treatment (P < 0.05). The digestibility of dry matter and protein was significantly increased (P < 0.05), while the digestibility of neutral detergent fiber, acid detergent fiber, and organic matter showed a numerical increase. Diets containing different levels of the Alternanthera sessilis plant did not affect pH, but the increase in the amount of this plant in the diet led to an increase ammonia nitrogen concentration and rumen protozoa population (P < 0.05). The addition of Alternanthera sessilis to the diet significantly reduced the concentration of blood glucose and cholesterol and increased the concentration of blood urea nitrogen (P < 0.05). Except for the weight of the thigh and neck, the effect of experimental treatments on other carcass components was not significant. Overall, the results of this experiment showed that using the Alternanthera sessilis plant in the rations of fattened lambs as a substitute for alfalfa forage not only had no negative effect on the studied parameters but also improved them in some cases. Therefore, Alternanthera sessilis can be used in rations of fattened lambs as a substitute for part of alfalfa forage.

Multi-omics integration analysis reveals the molecular mechanisms of drought adaptation in homologous tetraploid alfalfa(Medicago sativa 'Xinjiang-Daye').

Drought stress is a predominant abiotic factor leading to decreased alfalfa yield. Genomic ploidy differences contribute to varying adaptation mechanisms of different alfalfa cultivars to drought conditions. This study employed a multi-omics approach to characterize the molecular basis of drought tolerance in a tetraploid variant of alfalfa (Medicago sativa, Xinjiang-Daye). Under drought treatment, a total of 4446 genes, 859 proteins, and 524 metabolites showed significant differences in abundance. Integrative analysis of the multi-omics data revealed that regulatory modules involved in flavonoid biosynthesis, plant hormone signalling transduction, linoleic acid metabolism, and amino acid biosynthesis play crucial roles in alfalfa adaptation to drought stress. The severity of drought led to the substantial accumulation of flavonoids, plant hormones, free fatty acids, amino acids, and their derivatives in the leaves. Genes such as PAL, 4CL, CHI, CHS, PP2C, ARF_3, and AHP_4 play pivotal regulatory roles in flavonoid biosynthesis and hormone signalling pathways. Differential expression of the LOX gene emerged as a key factor in the elevated levels of free fatty acids. Upregulation of P5CS_1 and GOT1/2 contributed significantly to the accumulation of Pro and Phe contents. ERF19 emerged as a principal positive regulator governing the synthesis of the aforementioned compounds. Furthermore, observations suggest that Xinjiang-Daye alfalfa may exhibit widespread post-transcriptional regulatory mechanisms in adapting to drought stress. The study findings unveil the critical mechanisms by which Xinjiang-Daye alfalfa adapts to drought stress, offering novel insights for the improvement of alfalfa germplasm resources.

Effects of protein grass hay as alternative feed resource on lamb's fattening performance and meat quality.

Protein grass hay (PGH) was used as a new feed source for lambs to study its effect on fattening performance and meat quality. Fifty-six male lambs were allotted to four experimental groups and fed for eight weeks either alfalfa hay (AH)-based diet (control) or diets in which AH was replaced with 33 %, 66 %, or 99 % PGH. The inclusion of PGH did not affect final body weight, dry matter intake, average daily gain, feed conversion ratio, or carcass weight. Moreover, substituting AH with PGH at any level did not influence the ruminal fermentation or serum biochemical parameters, meat color, water holding capacity, shear force, or amino acid profile. However, relative liver weight was increased with 66 % substitutions. Furthermore, replacing 99 % AH with PGH decreased the meat's pH at 24 h. Higher levels of C18:3n-3, C20:5n-3, and total n-3 PUFA and a lower ratio of n-6: n-3 PUFA were also observed in meat from lambs fed PGH at 99 %. These findings suggest that PGH could be incorporated into the lamb's diet up to 99 % without compromising fattening performance and body health while improving their meat n-3 PUFA deposition.

Genome-wide investigation of the TIFY transcription factors in alfalfa (Medicago sativa L.): identification, analysis, and expression.

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.

Genome-wide investigation of the PIF gene family in alfalfa (Medicago sativa L.) expression profiles during development and stress.

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.

RNA-seq analyses on gametogenic tissues of alfalfa (Medicago sativa) revealed plant reproduction- and ploidy-related genes.

BACKGROUND: In alfalfa (Medicago sativa), the coexistence of interfertile subspecies (i.e. sativa, falcata and coerulea) characterized by different ploidy levels (diploidy and tetraploidy) and the occurrence of meiotic mutants capable of producing unreduced (2n) gametes, have been efficiently combined for the establishment of new polyploids. The wealth of agronomic data concerning forage quality and yield provides a thorough insight into the practical benefits of polyploidization. However, many of the underlying molecular mechanisms regarding gene expression and regulation remained completely unexplored. In this study, we aimed to address this gap by examining the transcriptome profiles of leaves and reproductive tissues, corresponding to anthers and pistils, sampled at different time points from diploid and tetraploid Medicago sativa individuals belonging to progenies produced by bilateral sexual polyploidization (dBSP and tBSP, respectively) and tetraploid individuals stemmed from unilateral sexual polyploidization (tUSP). RESULTS: Considering the crucial role played by anthers and pistils in the reduced and unreduced gametes formation, we firstly analyzed the transcriptional profiles of the reproductive tissues at different stages, regardless of the ploidy level and the origin of the samples. By using and combining three different analytical methodologies, namely weighted-gene co-expression network analysis (WGCNA), tau (τ) analysis, and differentially expressed genes (DEGs) analysis, we identified a robust set of genes and transcription factors potentially involved in both male sporogenesis and gametogenesis processes, particularly in crossing-over, callose synthesis, and exine formation. Subsequently, we assessed at the same floral stage, the differences attributable to the ploidy level (tBSP vs. dBSP) or the origin (tBSP vs. tUSP) of the samples, leading to the identification of ploidy and parent-specific genes. In this way, we identified, for example, genes that are specifically upregulated and downregulated in flower buds in the comparison between tBSP and dBSP, which could explain the reduced fertility of the former compared to the latter materials. CONCLUSIONS: While this study primarily functions as an extensive investigation at the transcriptomic level, the data provided could represent not only a valuable original asset for the scientific community but also a fully exploitable genomic resource for functional analyses in alfalfa.

Effects of temperature and lactic acid Bacteria additives on the quality and microbial community of wilted alfalfa silage.

This study investigated the influence of different temperatures (35℃ High temperature and average indoor ambient temperature of 25℃) and lactic acid bacterial additives (Lactiplantibacillus plantarym, Lentilactobacillus buchneri, or a combination of Lactiplantibacillus plantarym and Lentilactobacillus buchneri) on the chemical composition, fermentation quality, and microbial community of alfalfa silage feed. After a 60-day ensiling period, a significant interaction between temperature and additives was observed, affecting the dry matter (DM), crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) of the silage feed (p < 0.05). Temperature had a highly significant impact on the pH value of the silage feed (p < 0.0001). However, the effect of temperature on lactic acid, acetic acid, propionic acid, and butyric acid was not significant (p > 0.05), while the inoculation of additives had a significant effect on lactic acid, acetic acid, and butyric acid (p > 0.05). As for the dynamic changes of microbial community after silage, the addition of three kinds of bacteria increased the abundance of lactobacillus. Among all treatment groups, the treatment group using complex bacteria had the best fermentation effect, indicating that the effect of complex lactic acid bacteria was better than that of single bacteria in high temperature fermentation. In summary, this study explained the effects of different temperatures and lactic acid bacterial additives on alfalfa fermentation quality and microbial community, and improved our understanding of the mechanism of alfalfa related silage at high temperatures.

The potential of plant growth-promoting bacteria isolated from arid heavy metal contaminated environments in alleviating salt and water stresses in alfalfa.

Co-evolution of plant beneficial microbes in contaminated environments enhances plant growth and mitigates abiotic stress. However, few studies on heavy metal (HM) tolerant plant growth-promoting bacteria (PGPB) promoting crop growth in Morocco's farming areas affected by drought and salinity are available. Plant associated bacteria tolerant to HM and able to produce indole acetic acid and siderophores, display ACC-deaminase activity and solubilize phosphate, were isolated from long-term metal exposed environments. Tolerance to HM and biofilms formation in the absence or presence of HM were assessed. A consortium including two Ensifer meliloti strains (RhOL6 and RhOL8), one Pseudomonas sp. strain (DSP17), and one Proteus sp. strain (DSP1), was used to inoculate alfalfa (Medicago sativa) seedlings under various conditions, namely, salt stress (85 mM) and water stress (30% water holding capacity). Shoot and root dry weights of alfalfa were measured 60 days after sowing. In the presence of HM, DSP17 showed the greatest auxin production, whereas RhOL8 had the highest ACC-deaminase activity and DSP17 formed the densest biofilm. Root dry weight increased 138% and 195% in salt and water stressed plants, respectively, regarding non-inoculated controls. Our results confirm the improvement of alfalfa growth and mitigation of salt and drought stress upon inoculation.

Comparison of the effects of alfalfa meal and sorghum distillery residue supplementation on the methane emissions in black-feathered Taiwan native chicken.

The issue of global warming, primarily fueled by anthropogenic greenhouse gas emissions, necessitates effective strategies to address methane (CH4) emissions from both ruminants and nonruminants. Drawing inspiration from successful approaches employed in ruminants, this study evaluates the impact of supplementing the diets of Taiwan's native black-feathered chickens with alfalfa meal and sorghum distillery residues (SDRs) on CH4 emissions. Using a respiration chamber the results reveal a significant reduction in CH4 emissions when incorporating either 30% alfalfa meal or 30% SDRs into the chicken diet, demonstrating a 59% and 49% decrease, respectively, compared to the control group (P < 0.05). Considering that alfalfa meal contains saponins and SDRs contain tannins, the study delves into the mechanism through which these components mitigate CH4 production in chickens. Incorporating saponins or tannins shows that groups supplemented with these components exhibit significantly lower CH4 emissions compared to the control group (P < 0.05), with a consistent linear decrease as the concentration of the feed additive increases. Further in vitro analysis of chicken cecal contents indicates a proportional reduction in CH4 production with increasing levels of added saponins or tannins (P < 0.05). These findings suggest that the CH4-reducing effects of alfalfa meal and SDRs can be attributed to their saponins and tannin content. However, caution is warranted as excessive alfalfa meal supplementation may adversely impact poultry growth. Consequently, sorghum distillery residue emerges as a more suitable feed ingredient for mitigating CH4 emissions in Taiwan's native black-feathered chickens compared to alfalfa. Additionally, substituting SDRs for conventional commercial chicken feed not only reduces CH4 emissions but also enhances the utilization of byproducts.

In this study, researchers focused on addressing the crucial issue of global warming by tackling methane (CH4) emissions from chickens, an important source of greenhouse gases. We investigated the impact of adding alfalfa meal and sorghum distillery residues (SDRs) to the diets of Taiwan's native black-feathered chickens. The results showed a reduction in CH4 emissions when incorporating either 30% alfalfa meal or 30% SDRs into the chicken diet. This led to a 59% and 49% decrease, respectively, compared to the control group. The study explored the mechanism behind this reduction, pointing to the presence of saponins in alfalfa meal and tannins in SDRs as key components influencing CH4 production. However, the study also highlighted a potential concern regarding excessive alfalfa meal supplementation, which could negatively impact poultry growth. On the other hand, SDRs were found to have no detrimental effects, making them a more suitable feed ingredient for mitigating CH4 emissions in Taiwan's native black-feathered chickens compared to alfalfa. Moreover, substituting SDRs for conventional commercial chicken feed not only reduces CH4 emissions but also contributes to the enhanced utilization of byproducts, promoting a more sustainable approach to poultry farming.

Comparative proteomic discovery of salt stress response in alfalfa roots and overexpression of MsANN2 confers salt tolerance.

Soil salinity constrains growth, development and yield of alfalfa (Medicago sativa L.). To illustrate the molecular mechanisms responsible for salt tolerance, a comparative proteome analysis was explored to characterize protein profiles of alfalfa seedling roots exposed to 100 and 200 mM NaCl for three weeks. There were 52 differentially expressed proteins identified, among which the mRNA expressions of 12 were verified by Real-Time-PCR analysis. The results showed increase in abundance of ascorbate peroxidase, POD, CBS protein and PR-10 in salt-stressed alfalfa, suggesting an effectively antioxidant and defense systems. Alfalfa enhanced protein quality control system to refold or degrade abnormal proteins induced by salt stress through upregulation of unfolded protein response (UPR) marker PDIs and molecular chaperones (eg. HSP70, TCP-1, and GroES) as well as the ubiquitin-proteasome system (UPS) including ubiquitin ligase enzyme (E3) and proteasome subunits. Upregulation of proteins responsible for calcium signal transduction including calmodulin and annexin helped alfalfa adapt to salt stress. Specifically, annexin (MsANN2), a key Ca2+-binding protein, was selected for further characterization. The heterologous of the MsANN2 in Arabidopsis conferred salt tolerance. These results provide detailed information for salt-responsive root proteins and highlight the importance of MsANN2 in adapting to salt stress in alfalfa.