Search Results (179)

Alfalfa (Medicago sativa) persists for several years but must be rotated to another crop before replanting. Kura clover (T. ambiguum M. Bieb) is a perennial legume that can persist indefinitely without replanting; however, establishment is slow, which limits economic returns during the process. Two studies, each with four randomized complete blocks, were planted in two consecutive years at New Mexico State University’s Rex E. Kirksey Agricultural Science Center at Tucumcari, NM, USA, as the first known assessment evaluating alfalfa as an economic nurse crop during kura clover establishment using various kura clover–alfalfa drilled and broadcast planting arrangements. Irrigation termination due to drought limited yield measurements to three years after seeding. In that time, kura clover–alfalfa mixtures generally yielded equally to monoculture alfalfa, except for alternate row planting. After 5 years, the alfalfa stand percentage remained >80%, except for the alternate row treatment (69% stand). Kura clover monocultures attained about 40% stand, and the mixtures had a <25% stand. Alfalfa may persist for more than 5 years before relinquishing dominance to kura clover in mixtures, but the alfalfa would continue to provide economic returns as kura clover continues stand development with minimal production, but develops its root system to maximize production when released from the alfalfa nurse crop. Full article

Apomixis-mediated fixation of heterosis could transform hybrid breeding in alfalfa (Medicago sativa), a globally important forage crop. The parthenogenesis-inducing morphogenetic regulator BABY BOOM (BBM) represents a promising candidate for enabling this advancement. Here, we identified BBM homologs from three alfalfa genomes, characterized their promoter regions, and cloned a 2082 bp MsBBM gene encoding a 694-amino acid nuclear-localized protein. Three alfalfa BBM gene promoters primarily contained light- and hormone-responsive elements. Phylogenetic and conserved domain analyses of the MsBBM protein revealed a high sequence similarity with M. truncatula BBM. Expression profiling demonstrated tissue-specific accumulation of MsBBM transcripts, with the highest expression in the roots and developing pods. Hormonal treatments differentially regulated MsBBM. Expression was upregulated by GA₃ (except at 4 h) and SA, downregulated by NAA, MeJA (both except at 8 h), and ABA (except at 4 h), while ETH treatment induced a transient expression peak at 2 h. As an AP2/ERF family transcription factor showing preferential expression in young embryos, MsBBM likely participates in reproductive development and may facilitate apomixis. These findings establish a molecular framework for exploiting MsBBM to enhance alfalfa breeding efficiency through heterosis fixation. Full article

Vertical farming systems offer an efficient solution for sustainable food production in urban areas. However, managing nitrate (NO₃⁻) levels remains a significant challenge for improving crop yield, quality, and safety. This study evaluated the effects of nitrate availability on growth performance, nutrient uptake, and water use efficiency in a vertical hydroponic system that intercropped lettuce (Lactuca sativa) with alfalfa (Medicago sativa). The experiment was conducted in a controlled vertical hydroponic system using Nutrient Film Technique (NFT) channels, with nitrogen levels set at 0, 33, 66, 100, and 133% of the standard concentration. The results indicated that the intercropping treatment with 66% nitrate (IC-N_66%) improved water use efficiency by 38% and slightly increased leaf area compared to the other intercropping treatments. However, the control group, which consisted of a monoculture with full nitrate supply, achieved the highest overall biomass. Ion concentrations, including nitrate, calcium, magnesium, and micronutrients, were moderately affected by the intercropping strategy and nitrate levels. These findings suggest that moderate nitrate input, combined with nitrogen-fixing legumes, can enhance resource efficiency in hydroponic systems without significantly compromising yield. These findings offer a promising framework for incorporating legumes into hydroponic systems, minimizing the need for synthetic inputs while maintaining yield. These results support the use of agroecological intensification strategies in highly efficient soilless systems. Full article

In arid agricultural production, exploring suitable water-saving irrigation strategies and analyzing their water-saving mechanisms are of great significance. Alternating partial root-zone drying irrigation (APRI), a water-saving strategy, enhances the water use efficiency (WUE) of alfalfa (Medicago sativa L.) This paper aims to clarify the physiological mechanisms by which the APRI method enhances the physiological WUE of alfalfa, as well as the differences between this water-saving irrigation strategy, conventional irrigation (CI), and their water deficit adjustments, in order to seek higher water use efficiency for alfalfa production in arid regions. In this experiment, alfalfa was used as the research subject, and three irrigation methods, CI, fixed partial root-zone drying (FPRI), and APRI, were set up, each paired with three decreasing moisture supply gradients of 90% water holding capacity (WHC) (W1), 70% WHC (W2), and 50% WHC (W3). Samples were taken and observed once after every three complete irrigation cycles. Through a comparative analysis of the growth status, leaf water status, antioxidant enzyme activity, and osmotic adjustment capabilities of alfalfa under different water supplies for the three irrigation strategies, the following conclusions were drawn: First, the APRI method, through artificially created periodic wet–dry cycles in the rhizosphere soil, provides pseudo-drought stress that enhances the osmotic adjustment capabilities and antioxidant enzyme activity of alfalfa leaves during the early to middle phases of irrigation treatment compared to CI and FPRI methods, resulting in healthier leaf water conditions. Secondly, the stronger drought tolerance and superior growth conditions of alfalfa under the APRI method due to reduced water availability are key factors in enhancing the water use efficiency of alfalfa under this strategy. Full article

Background: Aluminum (Al) toxicity severely limits Medicago sativa (alfalfa) production on acidic soils, resulting in major yield losses worldwide. The highly conserved miRNA156 (miR156) functions by downregulating at least 11 SQUAMOSA promoter-binding protein-like (SPL) transcription factors in alfalfa, including SPL13, but its role in Al stress remains unclear. This study aimed to investigate the miR156/SPL regulatory network’s function in alfalfa under Al stress. Methods: Gene expression analyses, histochemical staining, nutrient profiling, phenotypic assays, transcriptome profiling, and ChIP-seq were conducted on alfalfa plants with altered miR156 and SPL13 expression to assess their roles in the Al stress response. Results: Al stress induced SPL13 expression while repressing miR156 in the roots. Elevated miR156 intensified Al accumulation, lipid peroxidation, and plasma membrane damage, accompanied by reduced leaf nitrogen, magnesium, sulfur, and phosphorus content. Phenotypically, increased SPL13 enhanced the root length and Al tolerance, whereas SPL13 silencing reduced tolerance. Transcriptome profiling of SPL13-silenced plants identified differentially expressed genes involved in the Al response, including aluminum-activated malate transporters and various transcription factors (GRAS, Myb-related, bHLH041, NAC, WRKY53, bZIP, and MADS-box). ChIP-seq revealed that SPL13 directly regulates genes encoding a protein kinase, cytochrome P450, and fasciclin-like arabinogalactan proteins. Conclusions: The MsmiR156/MsSPL13 network plays a crucial regulatory role in alfalfa’s response to Al toxicity. These findings provide novel genetic targets and foundational knowledge to advance molecular breeding for enhanced Al tolerance in alfalfa. Full article

The critical nitrogen dilution curve (CNDC) model enables precise nitrogen management by quantifying the threshold of nitrogen deficiency in crops, thereby enhancing both crop productivity and nitrogen use efficiency. However, its applicability to perennial crops remains unclear. In this study, alfalfa (Medicago sativa L.), a perennial leguminous forage, was used as the model crop. Based on two years of field experiments, CNDC models of aboveground biomass were constructed under two nitrogen fertilizer regimes: urea (0, 80, 160, and 240 kg·ha⁻¹, applied in a 6:2:2 basal-to-topdressing ratio) and controlled-release urea (CRU; 0, 80, 160, and 240 kg·ha⁻¹, applied as a single basal dose). Using these models, the nitrogen nutrition index (NNI) and cumulative nitrogen deficit (N_and) models were developed to diagnose alfalfa nitrogen status, and the optimal nitrogen application rates were determined via regression analysis. The results showed that critical nitrogen concentration and aboveground biomass followed a power function relationship under both fertilizer types. For CRU treatments, parameters a and b were 3.41 and 0.20 (first cut), 3.15 and 0.12 (second cut), and 2.24 and 0.40 (third cut), respectively. For urea treatments, a and b were 3.13 and 0.35 (first cut), 2.21 and 0.16 (second cut), and 1.75 and 0.73 (third cut). The normalized root mean square error (n-RMSE) of the models ranged from 3.1% to 13%, indicating high model reliability. Based on the NNI, N_and, and yield response models, the optimal nitrogen application rates were 175.44~181.71 kg·ha⁻¹ for urea and 145.63~153.46 kg·ha⁻¹ for CRU, corresponding to theoretical maximum yields of 14.76~17.40 t·ha⁻¹ and 16.76~20.66 t·ha⁻¹, respectively. Compared to urea, CRU reduced nitrogen input by 18.41~20.47% while achieving equivalent or higher theoretical yields. This study provides a scientific basis for nitrogen status diagnosis and precision nitrogen application in alfalfa cultivation. Full article

Alfalfa (Medicago sativa L.) is an important perennial leguminous forage; however, its high sensitivity to aluminum (Al) stress severely restricts its cultivation in regions with acidic soil. Therefore, this study conducted an integrated assessment of Al stress tolerance by performing systematic evaluations of 11 growth and physiological parameters across 30 alfalfa cultivars under Al stress, and calculated the Al tolerance coefficients based on these parameters. The results revealed that Al stress markedly inhibited root growth and biomass accumulation in alfalfa, thereby triggering increased malondialdehyde (MDA) content in roots across most cultivars, the scope of increase is 0.19–183.07%. Moreover, superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) increased by 7.50–121.44%, 2.50–135.89%, and 3.84–70.01%, respectively. Based on the comprehensive evaluation value (D) obtained via principal component analysis and membership function, the 30 alfalfa cultivars were categorized into four distinct groups: 4 highly Al-tolerant cultivars, 11 moderately high-Al-tolerant cultivars, 9 moderately low-Al-tolerant cultivars, and 6 low-Al-tolerant cultivars. Stepwise linear regression analysis identified root elongation rate, root-to-shoot ratio, root volume, SOD, MDA, CAT, root dry weight, POD, and root length as pivotal indicators for predicting and evaluating Al stress tolerance in alfalfa cultivars. The qRT-PCR analysis showed dynamic changes in ABC transporter gene expression in alfalfa roots over time under aluminum stress. Therefore, this study comprehensively evaluated Al tolerance by systematically investigating the morphophysiological effects of Al stress across 30 alfalfa cultivars using principal component analysis (PCA), membership function, and hierarchical clustering analysis. It provides a practical solution for expanding alfalfa planting in acid soil and improving feed production in acidic environments. Full article

Human agricultural activities can impact the soil microbial ecosystem, but the future implications of such changes remain largely unknown. This study aimed to explore how soil microbes survive and reproduce under the pressure of human agricultural cultivation and whether they resist or adapt. A 10-year continuous experiment was conducted, planting a maize and soybean rotation (control group), alfalfa (legume), and wheat (poaceae) to study the impact of different crop planting on soil microbial communities. During the experiment, the physical and chemical properties of the soil samples were measured, and the rhizosphere microbial communities were analyzed. Different crop plantings had varying effects on soil microbial species diversity, but these differences were relatively limited. The relative abundance of Cyanobacteriales (order) was higher in wheat than in alfalfa. Moreover, Cyanobacteriales were positively correlated with soil peroxidase, thereby promoting wheat growth. In addition, nutrition for fungi is mainly derived from decaying straw and plant roots. This study divided soil microbes under agricultural cultivation conditions into three categories: adaptive microbes, neutral microbes, and resistant microbes. At the ecological level of plant rhizosphere microbes, the plant rhizosphere soil microbial community showed a coevolutionary relationship with human cultivation activities. Future research needs to pay more attention to the adaptability of soil microbial communities to agricultural cultivation and the potential impact of this adaptability on the global ecosystem. Full article

Fusarium acuminatum is a major pathogenic fungus causing root rot in alfalfa (Medicago sativa). DNA polymerase theta is known to play a crucial role in repairing DNA double-strand breaks. However, its biological function in F. acuminatum remains unknown. In this study, the POLQ gene was deleted by homologous recombination using Agrobacterium tumefaciens-mediated transformation. Compared to the wild type (with the POLQ gene), the mutants (without the POLQ gene) showed significant phenotypic changes: they produced brown-yellow pigments instead of pink, slowed mycelial growth, and exhibited changes in macroconidia size and shape. The virulence of the mutants was greatly reduced, inducing only mild symptoms in alfalfa. In addition, FITC-WGA staining showed impaired spore germination and hyphal growth. These results suggest that POLQ is a key gene regulating growth and development of F. acuminatum, indicating that DNA repair may play an essential role in the pathogenicity of the pathogen in alfalfa. The POLQ gene could thus be a promising target for limiting F. acuminatum infections in alfalfa. Full article

The effects of stomatal factors of plant leaves under partial root-zone drying (PRD) have been widely studied. However, the non-stomatal factors and the relationship between photosynthesis with soil moisture have not been explored. In this study, four treatments over-irrigation, full irrigation, moderate water deficit, and severe water deficit were investigated, aiming to evaluate the effects on the diurnal variation of alfalfa leaf photosynthesis under PRD and its relationship with stomatal and non-stomatal limitations, as well as soil moisture. The results showed that any levels of water deficit led to a decrease in the photosynthetic rate (P_n) of alfalfa leaves. Leaves under moderate and severe water deficit displayed a pronounced midday “photosynthetic lunch break,” while those under over- and full irrigation did not display this phenomenon. Before 11:30 a.m., the reduction in P_n was primarily due to stomatal limitations, as evidenced by reduced stomatal conductance (G_s) and decreased intercellular CO₂ concentration (C_i). After 11:30 a.m., non-stomatal limitations became the dominant factor, with both G_s and transpiration rate (T_r) continuing to decrease, while C_i increased, indicating a shift in the limiting factors. Under PRD with moderate water deficit, alfalfa experienced both stomatal and non-stomatal limitations within a single day, leading to a hay yield reduction of 18.6%. Additionally, over-irrigation helped to maintain higher P_n and T_r, increasing alfalfa yield and thus improving water productivity by 33.1%. The correlation coefficients between soil moisture content at 10 cm depths with alfalfa leaf P_n, T_r, and G_s on the photosynthetic measurement day were 0.9864, 0.8571, and 0.8462, respectively. At 20 cm, the correlation coefficients were 0.8820, 0.6943, and 0.6951, respectively. The study concluded that both stomatal and non-stomatal mechanisms contributed to reduced alfalfa P_n in water deficit of PRD. Furthermore, shallow soil moisture also played a crucial role in influencing photosynthetic performance. Full article

Alfalfa is a deep-rooted perennial forage crop with diverse drought-tolerant traits. This study evaluated 250 alfalfa half-sib populations over three growing seasons (2021–2023) under irrigated and rainfed conditions in the Mediterranean drought-prone region of Central Chile (Cauquenes), aiming to identify high-yielding, drought-tolerant populations using remote sensing. Specifically, we assessed RGB-derived indices and canopy temperature difference (CTD; Tc − Ta) as proxies for forage yield (FY). The results showed considerable variation in FY across populations. Under rainfed conditions, winter FY ranged from 1.4 to 6.1 Mg ha⁻¹ and total FY from 3.7 to 14.7 Mg ha⁻¹. Under irrigation, winter FY reached up to 8.2 Mg ha⁻¹ and total FY up to 25.1 Mg ha⁻¹. The AlfaL4-5 (SARDI7), AlfaL57-7 (WL903), and AlfaL62-9 (Baldrich350) populations consistently produced the highest yields across regimes. RGB indices such as hue, saturation, b*, v*, GA, and GGA positively correlated with FY, while intensity, lightness, a*, and u* correlated negatively. CTD showed a significant negative correlation with FY across all seasons and water regimes. These findings highlight the potential of RGB imaging and CTD as effective, high-throughput field phenotyping tools for selecting drought-resilient alfalfa genotypes in Mediterranean environments. Full article

Drought stress is one of the key environmental factors restricting the germination of alfalfa seeds (Medicago sativa L.). Nanopriming is an innovative seed-priming technology able to meet economic, agronomic, and environmental needs in agriculture. However, the use of conventional nanomaterials is hampered by high costs, environmental risks, and biotoxicity. In this study, we synthesized iron oxide nanoparticles (Fe₃O₄NPs) using seasonal Ginkgo biloba leaf extracts (collected from August to November) obtained via an enzymatic ultrasonic-assisted method. The synthesized Fe₃O₄NPs were characterized using SEM, EDS, DLS, FTIR, UV-Vis, and XRD. To investigate the effects of Fe₃O₄NP priming on alfalfa seed germination under drought stress, germination and pot experiments were conducted with five Fe₃O₄NP priming concentrations (unprimed, 0, 20, 40, and 60 mg/L) and three PEG-6000 concentrations (0%, 10%, and 15%) to simulate normal, moderate, and severe drought conditions. The results showed that leaf extracts collected in November exhibited the highest flavonoid content (12.8 mg/g), successfully yielding bioactive-capped spherical Fe₃O₄NPs with a particle size of 369.5 ± 100.6 nm. Germination experiments revealed that under severe drought stress (15% PEG-6000), the 40 mg/L Fe₃O₄NP treatment most effectively enhanced seed vigor, increasing the germination rate, vigor index, and α-amylase activity by 22.1%, 189.4%, and 35.5% (p < 0.05), respectively, compared to controls. Under moderate drought stress (10% PEG-6000), the 20 mg/L Fe₃O₄NP treatment optimally improved germination traits, increasing the germination rate by 25.5% and seedling elongation by 115.6%. The pot experiments demonstrated morphological adaptations in alfalfa seedlings: under moderate drought stress, the 40 mg/L Fe₃O₄NPs significantly increased lateral root numbers, while under severe drought stress, the 60 mg/L Fe₃O₄NPs increased the root surface area by 20.5% and preserved the roots’ structural integrity compared to controls. These findings highlight that Fe₃O₄NPs synthesized via Ginkgo leaf extracts and enzymatic ultrasonic methods exhibit promising agricultural potential. The optimal Fe₃O₄NP priming concentrations enhanced seed vigor, germination traits, and drought resistance by modulating root morphology, with concentration-specific efficacy under varying drought intensities. Full article

Increasing soil salinity is threatening agricultural productivity which implies the development of new sustainable strategies to deal with this challenge. The main objective here is to assess the potential for improving the tolerance of alfalfa to salinity by combining inoculations with rhizobia and AMF. However, the distinguishing feature of this study is the comparison of two alfalfa varieties’ microbial response to salinity. The greenhouse trial was conducted on an Australian variety Siriver and an indigenous Demnate population, which were inoculated with Rhizoglomus irregulare and/or native AMF, and/or a RhOL1 rhizobial strain. The RhOL1 strain was selected from nine rhizobia tested for their plant growth promoting rhizobacteria (PGPR) activities. In addition to its ability to tolerate high salinity levels (769 mM) and solubilize insoluble phosphate as well as potassium, it can also synthesize auxins such as IAA. The application of these biofertilizers was carried out in the absence and the presence of the saline stress (0 and 120 mM NaCl). The double inoculations of native AMF and RhOL1 significantly improve the shoot and root dry biomass, plant elongation, number of formed leaves, and mineral nutrition, as well as the number of nodules and the rate of mycorrhizal root colonization. The synergistic effects between the native AMF and RhOL1 strain have been demonstrated in this study. However, the behavior of alfalfa genotypes towards microbial inoculation was significantly different. The ability to react to the double indigenous RhOL1 + AMF inoculation is more important in the Siriver than in the Demnate population. Thus, the possibility of formulating biofertilizers is based on the AMF–rhizobia–hote tripartite combination for alfalfa production in saline areas. Full article

Allelopathy plays a major role in agricultural production, influencing plant protection, crop yield, and crop rotation systems. This study investigated the effects of root exudates on 3105c alfalfa (Medicago sativa) seeds and seedlings to identify crops with strong and weak allelopathic potential. The results revealed that corn (Zea mays L.) (T1) exhibited the strongest allelopathic effects, whereas soybean (Glycine max (Linn.) Merr.) (T10) exhibited the weakest effects. T1 promoted seed germination by increasing radicle length and the simple vitality index. Both T1 and T10 promoted 3105c seedling growth and enhanced antioxidant capacity, albeit through different mechanisms. T1 primarily increased antioxidant capacity by elevating ascorbate and dehydroascorbate levels while reducing malondialdehyde content. In contrast, T10 enhanced antioxidant capacity by increasing soluble sugar and protein levels via hydroxyl free radical inhibition. These findings demonstrate that the allelopathic properties of corn effectively promote alfalfa growth by enhancing seed germination and improving physiological stress resistance. Full article

Mobile drip irrigation (MDI) systems integrate the technological advantages of center-pivot irrigation (CPI) systems and drip irrigation systems, boasting a high water-saving potential. To further enhance water use efficiency in alfalfa production in northern China, this preliminary study verified the accuracy of the HYDRUS-2D soil water movement numerical model through field experiments. Using the numerical model, four drip-line installation distances (60, 75, 90, and 105 cm), three deficit irrigation thresholds (45–50% FC, 55–60% FC, and 65–70% FC), and four irrigation depths (70% W, 85% W, 100% W, and 115% W) were set to simulate root water uptake, soil surface evaporation, total irrigation amount, and deep percolation during the entire growth cycle of alfalfa, respectively. Objective functions were constructed according to the simulation results, and the NSGA-II algorithm was used for multi-objective optimization of the deficit irrigation schedule. The preliminary results indicated that HYDRUS-2D can accurately simulate soil water movement under MDI systems, as the RMSE values of soil water content at all measured depths were less than 0.021 cm³/cm³, with the NRMSE values being below 23.3%, and the MAE values below 0.014 cm³/cm³. Increasing the deficit irrigation threshold from F1 to F3 enhanced root water uptake by 12.24–15.34% but simultaneously increased the total irrigation amount, soil surface evaporation (by up to 29.58%), and the risk of deep percolation; similar trends were observed with increasing irrigation depth. The drip-line installation distance had no significant impact on irrigation performance. The NSGA-II multi-objective optimization algorithm was used to obtain Pareto-optimal solutions that balance conflicting objectives. For this case study, a drip-line installation distance of 105 cm, a deficit irrigation threshold of 50–55% FC, and an irrigation depth of 112% W were recommended to achieve balance among the various optimization objectives. This study provides a preliminary framework for optimizing MDI systems and irrigation strategies. However, since a deeper root distribution (>80 cm) was not investigated in this study, future research incorporating deeper root zones is required for developing more comprehensive irrigation scheduling suitable for typical alfalfa cultivation scenarios. Full article