Clover Plants Research Articles

Optimizing polyphenol extraction and UPLC-MS/MS analysis from red clover (Trifolium pratense L.) species using response surface methodology

Polyphenols were extracted from red clover (Trifolium pratense L.) plants following conventional extraction. The combined factors of ethanol concentration (60–80%), extraction time (15–45 min) and extraction temperature (40–80°C) were evaluated. Central composite design software was used to conduct Response Surface Methodology (RSM) modelling and optimize the polyphenol yield from the red clover. Total phenolic content, antioxidant activity and ultra-performance liquid chromatography mass-spectrometry (UPLC-MS/MS) analysis were used to quantify polyphenols. The RSM model identified ethanolic composition having greatest influence (p < 0.05) on the polyphenol yield with extraction yields increasing with percentage ethanol. The optimal extraction conditions were identified as 80% ethanol for 45 minutes at 40°C. When compared with an exhaustive polyphenol extraction method (70% ethanol for 21 h at room temperature), the optimal extraction resulted in a 25.6% increase in TPC yield. UPLC-MS/MS was performed to quantify the individual polyphenols present within the sample, in which biochanin A and formononetin were present in abundance. The outcomes of this modelling and optimization study demonstrate that polyphenols from red clover can be extracted efficiently with minimal environmental and energy cost and thus had the capability to serve as a source of valuable nutraceutical and pharmaceutical products.

Contribution of mycorrhizal symbiosis and root strategy to red clover aboveground biomass under nitrogen addition and phosphorus distribution.

Soil nutrients exhibit heterogeneity in their spatial distribution, presenting challenges to plant acquisition. Notably, phosphorus (P) heterogeneity is a characteristic feature of soil, necessitating the development of adaptive strategies by plants to cope with this phenomenon. To address this, fully crossed three-factor experiments were conducted using red clover within rhizoboxes. Positions of P in three conditions, included P even distribution (even P), P close distribution (close P), and P far distribution (far P). Concurrently, N addition was two amounts(0 and 20mg kg- 1), both with and without AMF inoculation. The findings indicated a decrease in aboveground biomass attributable to uneven P distribution, whereas N and AMF demonstrated the potential to affect aboveground biomass. In a structural equation model, AMF primarily increased aboveground biomass by enhancing nodule number and specific leaf area (SLA). In contrast, N addition improved aboveground biomass through increased nodule number or direct effects. Subsequently, a random forest model indicated that under the far P treatment, fine root length emerged as the primary factor affecting aboveground biomass, followed by thickest root length. Conversely, in the even P treatment, the thickest root length was of paramount importance. In summary, when confronted with uneven P distribution, clover plants adopted various root foraging strategies. AMF played a pivotal role in elevating nodule number, and SLA.

Resistance Induction in Two Alfalfa Varieties to Bean Leafroll Virus and Citrus Excortis Viroid Using Some Botanicals and Amino Acids Preparations

Many researches have been conducted so far in the topic of controlling the Bean leafroll virus and Citrus exocortis viroid. This study was conducted in a private farm at Balad district from September 1, 2023, to January 6, 2024, and aimed to evaluate the efficacy of azolla and clover plant extract, fosetyl aluminum pesticide, and Debal against Bean leafroll virus in the presence of Citrus exocortis viroid in diseased alfalfa. The diagnosis was confirmed for the whole genome of the virus and viroid under study using next generation sequencing technology. The resulting sequences were compared in the International Gene Bank database NCBI. The virus was the 1st record registered in Iraq, which was registered with a accession number PP889325, and the viroid genome was registered for the first time in the NCBI database with an accession number PP869624. Findings of the field experiment showed that the treatments were effective in reducing infection rates and disease severity beside increasing plant defense enzymes (catalase, polyphenol oxidase, and peroxidase) in virus infected alfalfa. Debal treatment, individual or in combination, recorded the highest rate leaf content of catalase (8.00), polyphenol oxidase (3.70), and peroxidase (4.13), with values significantly differed from infected untreated control treatment that recorded 3.33, 0.91, and 0.81, respectively.

Adaptation of Rhizobium leguminosarum sv. trifolii strains to low temperature stress in both free-living stage and during symbiosis with clover

Legume-rhizobial symbiosis plays an important role in agriculture and ecological restoration. This process occurs within special new structures, called nodules, formed mainly on legume roots. Soil bacteria, commonly known as rhizobia, fix atmospheric dinitrogen, converting it into a form that can be assimilated by plants. Various environmental factors, including a low temperature, have an impact on the symbiotic efficiency. Nevertheless, the effect of temperature on the phenotypic and symbiotic traits of rhizobia has not been determined in detail to date. Therefore, in this study, the influence of temperature on different cell surface and symbiotic properties of rhizobia was estimated. In total, 31 Rhizobium leguminosarum sv. trifolii strains isolated from root nodules of red clover plants growing in the subpolar and temperate climate regions, which essentially differ in year and day temperature profiles, were chosen for this analysis. Our results showed that temperature has a significant effect on several surface properties of rhizobial cells, such as hydrophobicity, aggregation, and motility. Low temperature also stimulated EPS synthesis and biofilm formation in R. leguminosarum sv. trifolii. This extracellular polysaccharide is known to play an important protective role against different environmental stresses. The strains produced large amounts of EPS under tested temperature conditions that facilitated adherence of rhizobial cells to different surfaces. The high adaptability of these strains to cold stress was also confirmed during symbiosis. Irrespective of their climatic origin, the strains proved to be highly effective in attachment to legume roots and were efficient microsymbionts of clover plants. However, some diversity in the response to low temperature stress was found among the strains. Among them, M16 and R137 proved to be highly competitive and efficient in nodule occupancy and biomass production; thus, they can be potential yield-enhancing inoculants of legumes.

Classifying Stand Compositions in Clover Grass Based on High-Resolution Multispectral UAV Images

In organic farming, clover is an important basis for green manure in crop rotation systems due to its nitrogen-fixing effect. However, clover is often sown in mixtures with grass to achieve a yield-increasing effect. In order to determine the quantity and distribution of clover and its influence on the subsequent crops, clover plants must be identified at the individual plant level and spatially differentiated from grass plants. In practice, this is usually done by visual estimation or extensive field sampling. High-resolution unmanned aerial vehicles (UAVs) offer a more efficient alternative. In the present study, clover and grass plants were classified based on spectral information from high-resolution UAV multispectral images and texture features using a random forest classifier. Three different timestamps were observed in order to depict the phenological development of clover and grass distributions. To reduce data redundancy and processing time, relevant texture features were selected based on a wrapper analysis and combined with the original bands. Including these texture features, a significant improvement in classification accuracy of up to 8% was achieved compared to a classification based on the original bands only. Depending on the phenological stage observed, this resulted in overall accuracies between 86% and 91%. Subsequently, high-resolution UAV imagery data allow for precise management recommendations for precision agriculture with site-specific fertilization measures.

AllergenAI: a deep learning model predicting allergenicity based on protein sequence.

Innovations in protein engineering can help redesign allergenic proteins to reduce adverse reactions in sensitive individuals. To accomplish this aim, a better knowledge of the molecular properties of allergenic proteins and the molecular features that make a protein allergenic is needed. We present a novel AI-based tool, AllergenAI, to quantify the allergenic potential of a given protein. Our approach is solely based on protein sequences, differentiating it from previous tools that use some knowledge of the allergens' physicochemical and other properties in addition to sequence homology. We used the collected data on protein sequences of allergenic proteins as archived in the three well-established databases, SDAP 2.0, COMPARE, and AlgPred 2, to train a convolutional neural network and assessed its prediction performance by cross-validation. We then used Allergen AI to find novel potential proteins of the cupin family in date palm, spinach, maize, and red clover plants with a high allergenicity score that might have an adverse allergenic effect on sensitive individuals. By analyzing the feature importance scores (FIS) of vicilins, we identified a proline-alanine-rich (P-A) motif in the top 50% of FIS regions that overlapped with known IgE epitope regions of vicilin allergens. Furthermore, using∼ 1600 allergen structures in our SDAP database, we showed the potential to incorporate 3D information in a CNN model. Future, incorporating 3D information in training data should enhance the accuracy. AllergenAI is a novel foundation for identifying the critical features that distinguish allergenic proteins.

Evaluation of the influence of a new triazole derivative on the period vegetation and 1st phase of growth of creeping clover seeds white (Trifolium repens L.)

The use of heterocyclic compounds from a number of triazole derivatives is well known in the agricultural sector. For the most part, the fungicidal properties of this group of chemical compounds allowed the occupied niche of one of the most successful in the use of antifungal drugs. However, previous research revealed that these compounds also affect the growth-regulatory indicators of plants, which in turn was demonstrated in our work. In this work, the results of the influence of three different concentrations of a promising triazole derivative compound on the effect on the growth of the vegetative and generative parts of the meadow clover plant germinated from seeds during the vegetation period and the first phase of growth were investigated. The research results showed that creeping (white) clover seeds germinate very quickly – germination was observed already the next day in all variants, and after three days this indicator ranged from 55 to 64%. We obtained the lowest results in variants with the lowest concentrations of the growth regulator – 93% compared to the control. We obtained the highest germination in the version with a solution of the experimental growth regulator at a concentration of 0.02% – the number of germinated seeds is 8.5% more than in the control. A higher concentration of this substance (0.04%) did not show an improvement in germination – the number of germinated clover seeds in this variant was almost no different from the control. Our results indicate that the triazole derivative in the studied concentrations affects the vegetation period and the 1st growth phase of creeping white clover seeds. At the same time, it has a slight stimulating effect in low concentrations, and also protects against the negative impact of soil infections.

In Vitro Screening of Endophytic Micromonospora Strains Associated with White Clover for Antimicrobial Activity against Phytopathogenic Fungi and Promotion of Plant Growth

Bacteria belonging to the genus Micromonospora are recognized as microorganisms with the potential to be used in biotechnology processes, given their beneficial influence on plant growth and the biocontrol of phytopathogens. In this study, nineteen Micromonospora isolates originating from the root nodules of white clover plants were taxonomically assigned based on the phylogenetic analysis of the 16S rRNA gene and four housekeeping genes. The antifungal properties of the bacteria against phytopathogenic Botrytis cinerea, Fusarium oxysporum, Fusarium equiseti, Sclerotinia sclerotiorum, and Verticillium albo-atrum were tested with the agar plug test and the dual culture test. The ability to produce various metallophores was determined with the agar plug diffusion test on modified chrome azurol S (CAS) agar medium. International Streptomyces Project-2 medium (ISP2) broth amended with 0.2% L-tryptophan was used to indicate the bacterial ability to produce auxins. The strains belonging to M. tulbaghiae, M. inaquosa, and M. violae showed in vitro potential as antimicrobial agents against the tested fungi. M. inaquosa strain 152, M. violae strain 126, M. violae strain 66, and M. violae strain 45 were recognized as the most efficient metallophore producers. M. alfalfae strain 55 and M. lupini strain 5052 were identified as the most promising auxin compound producers and, therefore, show potential as plant-growth-promoting bacteria.

Genome assembly of Melilotus officinalis provides a new reference genome for functional genomics

BackgroundSweet yellow clover (Melilotus officinalis) is a diploid plant (2n = 16) that is native to Europe. It is an excellent legume forage. It can both fix nitrogen and serve as a medicine. A genome assembly of Melilotus officinalis that was collected from Best corporation in Beijing is available based on Nanopore sequencing. The genome of Melilotus officinalis was sequenced, assembled, and annotated.ResultsThe latest PacBio third generation HiFi assembly and sequencing strategies were used to produce a Melilotus officinalis genome assembly size of 1,066 Mbp, contig N50 = 5 Mbp, scaffold N50 = 130 Mbp, and complete benchmarking universal single-copy orthologs (BUSCOs) = 96.4%. This annotation produced 47,873 high-confidence gene models, which will substantially aid in our research on molecular breeding. A collinear analysis showed that Melilotus officinalis and Medicago truncatula shared conserved synteny. The expansion and contraction of gene families showed that Melilotus officinalis expanded by 565 gene families and shrank by 56 gene families. The contacted gene families were associated with response to stimulus, nucleotide binding, and small molecule binding. Thus, it is related to a family of genes associated with peptidase activity, which could lead to better stress tolerance in plants.ConclusionsIn this study, the latest PacBio technology was used to assemble and sequence the genome of the Melilotus officinalis and annotate its protein-coding genes. These results will expand the genomic resources available for Melilotus officinalis and should assist in subsequent research on sweet yellow clover plants.

Influence of flowering red clover on flower visitation in a sweet corn agroecosystem

Agricultural intensification and the conversion of natural landscapes into annual cropping systems have contributed to declines in pollinator abundance and biodiversity. Increasing the abundance of flowering plants within crop fields is an often-overlooked practice that may be used to help sustain and enhance pollinator populations. In this study, the influence of red clover (Trifolium pratense) used as an interplanted living mulch on pollinator richness and visitation rates was evaluated and compared with monoculture sweet corn habitats. Treatments included sweet corn interplanted with red clover or monoculture with or without cover crop residue. Weekly visual observations of foraging floral visitors revealed that multiple species of bumblebees and butterflies, as well as honeybees frequently visited red clover flowers. Observations of visitors foraging on sweet corn tassels during pollen shed revealed distinct insect communities were attracted by sweet corn and red clover plants. Findings provided evidence that the inclusion of red clover in crop fields can increase the diversity and abundance of bees and butterflies on arable lands by serving as an important food source.

Effects of flue gas desulfurization gypsum and clover planting on qualities of soil and winter jujube in coastal saline-alkali orchard of north China

Effects of flue gas desulfurization gypsum and clover planting on qualities of soil and winter jujube in coastal saline-alkali orchard of north China

ASSESSMENT OF HEAVY METALS ACCUMULATION IN PERENNIAL LEGUMES (WHITE CLOVER AND RED CLOVER) IN THE COPȘA MICĂ AREA, ROMANIA

The impact of heavy metals on perennial legumes, just like on other plant species, can be detrimental and has several ecological and agricultural implications. The ability of perennial legumes to fix atmospheric nitrogen through symbiotic interactions with nitrogen-fixing bacteria in their root nodules makes them a significant group of plants. The present study aims to evaluate the total concentration of heavy metals (cadmium, lead, copper, and zinc) in red and white clover plants harvested from grasslands located in heavily polluted areas (Copșa Mică, Axente Sever, Valea Viilor, Micăsasa and Târnava). White clover (Trifolium repens L.) recorded Cd content values between 0.06 - 3.53 mg/kg dry weight, Cu had values between 4.09 - 33.4 mg/kg dry weight, Pb content was between 0.18 - 4.77 mg/kg dry weight and Zn had values between 38 - 186 mg/kg dry weight. Regarding red clover (Trifolium pratense L.) the heavy metal content values were as follows: for Cd (0.08 - 3.38 mg/ kg d.w), for Cu (6.03 - 13.5 mg/kg d.w), for Pb (0.13 – 2.34 mg/kg d.w), for Zn (33.1 - 169 mg/kg d.w). Our results show that clover species harvested from grasslands in these geographical areas have high levels of heavy metals due to metallurgical activities. Grazing in these areas affected by heavy metal pollution can pose serious health problems for animals and later for humans through consumption of their products.

Influence of cultivation techniques on productivity and quality of meadow clover

Meadow clover (red clover) is the most important fodder plant in most parts of Russia. Positive characteristics of this plant are the presence of a significant amount of reserve nutrients (crude protein, protein, nitrogenfree extractive substances) in the composition of plant tissues, which provides high fodder value. As a legume, meadow clover accumulates in the soil organically bound nitrogen, a large amount of organic mass of crop root residues, enriching the soil with nutrients and contributing to the increase of its physical and mechanical properties. The disadvantage of this crop is its short productive life. In this regard, it is necessary to intensify the cultivation methods for a short period of use in order to obtain the greatest return from the crop in the fodder and agronomic aspect. Intensification of cultivation methods means everything that contributes to the increase of plant productivity: application of mineral and organic fertilizers to improve mineral nutrition, inoculation of seeds before sowing to mobilize and activate soil microflora and productive symbiosis between plants and microbiota, use of growth regulators to optimize photosynthesis processes. The study of the influence of these intensification methods was the purpose of our research. It was found that the use of estimated doses of mineral fertilizers contributed most optimally to the processes of yield formation and final productivity of clover grass. Among all the strains of nodule bacteria studied, inoculant strain 348a was the most effective because it is speciesspecific for meadow clover. The use of growth regulators Ribav Extra and Root Super is a positive technique that provides optimal growth and development of plants during vegetation. The methods studied also had a positive effect on the seed productivity of the clover plants, increasing yield and seed quality.

Urban spatial heterogeneity shapes the evolution of an antiherbivore defense trait and its genes in white clover

Urbanization is a global threat to biodiversity due to its large impact on environmental changes. Recently, urban environmental change has been shown to impact the evolution of many species. However, much remains unknown about how urban environments influence evolutionary processes and outcomes due to the non‐linearity and discontinuity of environmental variables along urban–rural gradients. Here, we focused on the evolution of hydrogen cyanide (HCN) production and its components (presence/absence of cyanogenic glycosides and the hydrolytic enzyme linamarase) in the herbaceous plant white clover Trifolium repens, which thrive in both urban and rural areas. To comprehensively elucidate how plants evolve and adapt to heterogenous urban environments, we collected 3299 white clover plants from 122 populations throughout Sapporo, Japan. We examined the spatial variation in environmental factors, such as herbivory, sky openness, impervious surface cover, snow depth, and temperature, and how variation in these factors was related to the production of HCN, cyanogenic glycosides, and linamarase. Environmental factors showed complex spatial variation due to the heterogeneity of the urban landscape. Among these factors, herbivory, sky openness, and impervious surface cover were highly related to the frequency of plants producing HCN in populations. We also found that impervious surface cover was related to the frequency of plants producing cyanogenic glycosides, while herbivory pressure was not. As a result, the cyanogenic glycoside frequency showed a clearer trend along urban–rural gradient rather than HCN frequency, and thus, the predicted spatial distributions of HCN and cyanogenic glycosides were inconsistent. These results suggest that urban landscape heterogeneity and trait multifunctionality determines mosaic‐like spatial distribution of evolutionary traits.

Effect of mixing ratios and spraying with microelements on the growth and yield of forage for a mixture of oats and clover

A field experiment was carried out in the fields of the Agricultural Research Station-College of Agriculture, University of Basra, during the winter season of 2021-2022. To study the effect of seeding rates of both oats and Egyptian clover with mixing percentages of 100% oats, 75% oats + 25% clover, 50% oats + 50% clover, 75% clover + 25% oats, 100% clover as the first factor and microelements at a concentration of 1500, 3000 and 4500 ppm as a second factor. The time for spraying with micro-elements was in the branching stage, 7-10 days after the first spray and 7-10 days after the second spray. The experiment was carried out in a factorial experiment with three replications, according to a randomized complete block design, RCBD. Three cuttings were taken during the crop’s growth period. The height of the oats and clover plants, the number of oat tillers, the number of clover branches, the weight ratio of leaves to tillers and stems, and the amount of green and dry forage were measured. The results of the study showed that the different ratios of mixing oats and clover crops contributed to the production of the studied traits; 50% of oats + 50% of clover in the second cutting recorded the highest yield of green fodder (66.84 tons ha-1) and dry fodder yield (22.37 tons ha-1). As for the effect of concentrations of microelements, the level of 4500 ppm achieved superiority in all growth characteristics of all cutting, and the highest yield was recorded in green fodder (65.57 tons ha-1) and dry forage (22.44 tons ha-1) at the second cutting. 50% oats + 50% clover at the level of 4500 ppm gave the highest yield for green fodder (69.66 tons ha-1) and dry (24.81 tons ha-1). Keywords: Mixture; Spraying; micro-nutrients; green fodder; Avena sativa; Clover.

Effects of Drought Stress on Red Clover-Grass Mixed Stands Compared to Grass Monoculture Stands in Nitrogen-Deficient Systems

Drought stress is one of the major abiotic stress factors affecting forage production; thus, it is essential to obtain a better understanding of how forage responds to drought. The main objective of this study was to evaluate how legume-grass mixed forage stands respond to drought stress when compared to grass monoculture. A greenhouse pot experiment was conducted using a red clover (Trifolium pratense L.)—timothy grass (Phleum pratense L.) mixed stand and a timothy monoculture stand, where plants were subjected to severe drought (20% field capacity—FC), moderate drought (40% FC), and well-watered (80% FC) conditions for four weeks and subsequently allowed to recover for another four weeks by adjusting moisture back to 80% FC. Both moderate and severe droughts significantly reduced the shoot biomass of the mixed stand, while no difference was exhibited in the timothy monoculture. The shoot biomass and nitrogen fixation capacity of red clover were reduced under drought stress. However, red clover plants subjected to moderate drought were able to recover shoot growth and nitrogen fixation capacity during the recovery phase, allowing more biologically fixed nitrogen and shoot nitrogen production similar to the plants growing under well-watered conditions. Overall, the results demonstrate that the inclusion of legumes in forage mixtures enhances resilience to moderate drought stress.

Isolation and Molecular Diagnosis of Enterobacter cloacae and Kineococcus radiotolerans from Red Clover nodules and evaluation of the prepared inoculum from them as a stimulator for plant growth

To obtain biological inoculum that promotes plant growth and contributes to a clean environment and sustainable agriculture, twentyone samples were collected from the root nodes of the red clover plant (Trifolium paratens) grown in gypsiferous soils at the Research Station of the Department of Soil Sciences and Water Resources - Faculty of Agriculture - University of Tikrit, samples were cultured in the medium of YEMA. Two isolates belonging to Enterobacter cloacae and Kineococcus radiotolerans were diagnosed phenotypically and molecularly. The diagnosis was confirmed by analyzing the nitrogen bases sequence of the DNA of the 16S rRNA gene. It was recorded in the National Center Biotechnology Information(NCBI) with the numbers MN310027.1 and KT216573.1 respectively, these isolates were activated and tested their efficiency as a bio-stimulant by testing their ability to dissolve insoluble phosphate compounds and produce indole acetic acid and chelating compounds, the results showed the ability of all isolates to produce indole acetic acid (IAA), chelating compounds, and solubility of phosphates, E. cloacae isolate outperformed K. radiotolerans isolates in these parameters, as it gave E. coloaca solubilization of phosphate and IAA production of 40.3 mg p L-1 and 11.2 µg ml-1 respectively, and it showed a high output of the chelating compounds compared to the treatment inoculated with K. radiotolerans, which gave 27.20 mg p L-1 and 7.21 µg ml-1, and medium production of iron chelating compounds, the results also showed the superiority of the inoculated treatments over the uninoculated treatments in the percentage of germination, the speed of germination, the length of tomato seedlings, the dry weight of the Shoot and root parts and the number of leaves, and the results showed the superiority of the inoculated treatment with E. cloacae significantly on K. radiotolerans isolate. Keywords: Enterobacter cloacae; Kineococcus radiotolerans; Molecular Diagnosis; Bacterial inoculation; Eggplant seedlings.

The Effect of Tillage Systems, Type of Organic Fertilizer and the Level of Binary Addition of Nitrogen and Phosphorus on the Growth and Yield of Clover in Gypsum Soil

A field experiment was conducted at the research station of the College of Agriculture - University of Tikrit for the agricultural season (2021-2022), and the experiment factors were three, namely: the first factor is tillage systems, as two systems of tillage were applied (no-tillage and Reduced tillage) and the second factor was the type of organic fertilizer and it included (Sheep waste and waste Mushroom), while the third factor was the level of binary addition of nitrogen and phosphorus, and it included three levels of fertilization (0%, 100%, and 125%), according to the fertilizer recommendation. The experiment was carried out according to the split-split plot system with a randomized complete block design (R.C.B.D) and with three replications. The treatments were distributed as follows: tillage systems (NT, RT) in the main plot, and the application of the type of organic fertilizer (O1, O2) In the sub plot, the levels of binary addition of nitrogen and phosphorus (M0, M1, M2) were placed in the sub-sub plots, and the results of the experiment showed that the system of Reduced tillage was significantly superior to the system of no-tillage in all the studied traits, and sheep waste was significantly superior to mushroom waste in All the characteristics of the study, as well as the level of binary addition M2 was significantly superior to the other levels in all the characteristics of the study. The results showed that the treatment (T1O1M2) was significantly superior in all traits of vegetative growth and yield traits of clover plant.

Impact of Mulching on Soil Moisture and Sap Flow Characteristics of Jujube Trees

The main purpose of this study was to assess the influence of grass planting and jujube branch mulching on soil moisture levels and jujube tree transpiration rates, with the ultimate goal of improving jujube tree production in rain-fed orchards. The study encompassed four treatments: jujube branch mulching (JBM), jujube branch mulching with white clover planting (JBM + WCP), white clover planting (WCP), and clean cultivation (CC). During a two-year experiment, it was observed that the JBM treatment exhibited the highest capacity for moisture conservation. Specifically, it resulted in an average increase of 2.69% (in 2013) and 2.23% (in 2014) in soil moisture content compared with the CC treatment. The application of statistical analysis revealed significant differences (p &lt; 0.05) between JBM and JBM + WCP, as well as highly significant differences (p &lt; 0.01) between JBM and WCP in the year 2013. In 2014, JBM exhibited significant differences (p &lt; 0.01) from both JBM + WCP and WCP. Between April and August, JBM exhibited the highest soil moisture content, followed by CC, with WCP showing the lowest levels. From September to October, JBM retained its status as the treatment with the highest soil moisture content, JBM + WCP ranked second, and CC experienced a decline and recorded the lowest soil moisture content. Under sunny conditions, all treatments showed a broad peak curve in the daily variation of sap flow velocity. In cloudy weather, a multi-peak wave-like curve was observed with similar trends across treatments. Between April and August, the monthly average sap flow velocity of JBM ranked the highest, followed by CC, while WCP showed the lowest velocity. During the period of September to October, JBM maintained its lead in sap flow velocity, while JBM + WCP rose to the second position, and CC’s sap flow velocity dropped to the lowest level. JBM and WCP treatments showed significant differences (p &lt; 0.01), and in 2014, JBM also had significant differences (p &lt; 0.05) compared with JBM + WCP. The sap flow velocity was positively correlated with air temperature, vapor pressure deficit, wind velocity, photosynthetically active radiation, and soil temperature. Photosynthetically active radiation was identified as the main driving factor influencing jujube tree transpiration. In conclusion, the findings of this study demonstrate the effectiveness of using pruned jujube branches for coverage in rain-fed jujube orchards. This approach not only conserves mulching materials and diminishes the expenses associated with transporting pruned jujube tree branches away from the jujube orchard but also achieves multiple objectives, including increasing soil moisture, promoting jujube tree transpiration, and enhancing soil water utilization. These results have significant implications for the efficient utilization of rainwater resources in rain-fed jujube orchards and provide valuable insights for practical applications in orchard management.

A robust hydroponic‐based system for screening red clover (Trifolium pratense) for Fusarium avenaceum

AbstractRed clover (Trifolium pratense) is an important forage legume crop that suffers like most perennial crops from attacks by soil‐borne pathogens. Fusarium root rot is one of the most serious diseases and at the same time problematic to identify resistance because of its hidden life in the soil. Current screening methods are laborious and hampered by limited reproducibility. To remedy this situation, we aimed to establish a simple and reliable hydroponics‐based screening system to facilitate studies of red clover–Fusarium avenaceum interactions. First, the fungal spore concentrations were balanced toward the development of red clover plants grown hydroponically. We found that the optimum concentration was 30,000 spores in 2 L of hydroponic medium to ensure infection during the plant growth period in this system. The procedure was scaled‐up to screen plants from 25 populations to identify red clover individuals with the improved resistance to F. avenaceum. Susceptible plants had approximately two‐fold higher amounts of fungal DNA than resistant plants, demonstrating a correlation between the disease readings of the plants and pathogen DNA. We foresee this screening procedure meeting the needs of both applied breeding work and in‐depth molecular studies of responses between this pathogen and its host plant. This method could be applied for the screening of other plant species for resistance to Fusarium spp. or to other root microbes.