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The presented study deals with the use of legumes intercropped with maize for the production of biogas from silage. The main goal was to find out whether silages made from mixed cultures can be used in biogas production and how the use of such silages affects qualitative and quantitative parameters of the fermentation process compared with the pure maize silage. Variants prepared were pure cultures of maize, bean, lupin, and white sweet clover. In addition, mixed cultures were prepared of maize and individual legumes. Measured values showed that in terms of dry matter (DM) yield, mixed culture silages are almost of the same or even better quality than silage made from the maize monosubstrate. Compared with the maize monoculture silage, the presence of white lupine, white sweet clover, and broad bean in silages statistically significantly increased the content of DM, ash, and acid detergent fiber (by more than 5%). Bean and lupine in mixed silages with maize significantly increased the content of lipids (on average by more than 1.2%). Legumes in silages were significantly decreasing contents of neutral detergent fiber, crude protein, and starch. Production of biogas from silages of maize monosubstrates and mixed substrates of maize with white lupin, maize with white sweet clover, and maize with broad bean was directly proportional to the content of CAR and starch in these substrates. A perspective variant was the mixed substrate of maize and sweet clover from which biogas production was only 6% lower than that from conventional maize silage. The highest yield was recorded in the maize monosubstrate (0.923 m3/kgVS). Variants of mixed substrates had a yield ranging from 0.804 to 0.840 m3/kgVS. [ABSTRACT FROM AUTHOR]

Background. Dairy farming is one of the main segments in the agricultural sector of Murmansk Province. Expanding the range of cultivated legume fodder crops and their cultivars is one of the ways to comprehensively solve the problem of feed in the Kola Peninsula. Introduction of perennial legumes into the feed production practice will reduce the production cost of feeds and improve their quality.Materials and methods. Fodder galega, variable and yellow alfalfa (2005–2009), bird’s-foot trefoil (2006–2007), and sweet clover (2007–2008) were studied at the Polar Experiment Station of VIR. Weather conditions during the years of study differed in temperature and water regimes. Observations and records were made according to VIR’s guidelines. Statistical data processing included the calculation of the main variation parameters and the construction of a generalized linear model of the variance structure.Results. The data are presented on the study of the collection of four perennial fodder legumes in Murmansk Province, Russia, in 2005–2009. Alfalfa surpassed fodder galega, sweet clover and bird’s-foot trefoil in winter hardiness, height and bushiness, showed medium leafiness, and (for the entire study cycle) had low air-dry mass yield. Biennial bicarpic sweet clover stood out for its air-dry mass yield on average per year, had medium height and foliage, but low winter hardiness and bushiness. Bird’sfoot trefoil was significantly inferior to the rest of the studied crops in all agronomic characteristics. Galega had the highest leafiness; in winter hardiness, bushiness and plant height it was slightly inferior to alfalfa, and in yield to white clover. Seeds under the conditions of Murmansk Province can be obtained only from galega accessions.Conclusion. According to a set of agronomic characters, the most promising crop is fodder galega, especially its adapted naturalized accessions selected in 1991–1993.

In this paper, a possible use of white sweet clover (Melilotus albus Med.) for phytoremediation was assessed. The plants were grown on soils with naturally occurring concentrations of potentially toxic elements (PTEs). First, the content of PTEs was determined in plant biomass and in soil samples using: (a) Optical emission spectrometry with inductively coupled plasma to determine Sb, As, Cd, Cu, Ni, Pb, and Se, and (b) thermal decomposition, amalgamation, and atomic absorption spectrometry to determine Hg. The effectiveness of Melilotus albus Med. (M. albus) for phytoremediation was evaluated using the bioconcentration factor (BCF). The phytoextraction potential of M. albus was determined using bioaccumulation factor (BAC) and translocation factor (TF) values. The highest concentration of PTEs in roots was detected for zinc (10.56 mg/kg of dry weight, DW) and copper (5.128 mg/kg of DW). Similarly, the highest concentration in above-ground parts of the plant was detected for zinc and copper (12.638 and 4.0 mg/kg of DW, respectively). Although the values were relatively high, the effectiveness of the absorption of these PTEs by plant biomass from the soil was relatively very low. BAC and BCF were always lower than 1. On the other hand, the results suggested that M. albus effectively transports PTEs (only for Zn, Pb and Hg) from roots to shoots, because TF was always higher than 1. However, the accumulation of PTEs from soils with a natural abundance of PTEs was not excessive in comparison to conventional maize silage. Therefore, there is no potential risk of biomethane production in biogas plants when biomass from M. albus is used. [ABSTRACT FROM AUTHOR]

Identifying sustainable resources of natural fibers is essential due to their high demand in industrial applications such as automotive and biomedical materials. Two alternative fibers obtained from canola and sweet clover stalks were characterized for their properties using energy dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), contact angle, and tensile test. Hemp and flax fibers, both in use as industrial fibers, were also characterized as conventional fibers. Results showed that all the fibers had the same chemical elements (carbon, oxygen, magnesium, and potassium) and chemical bonds. The crystallinity index for the alternative fibers ranged from 62 to 71%, which was close but lower than the conventional fibers (82% for hemp and 80% for flax). The thermal stability of the alternative fibers was around 220 °C, close to the conventional fibers (230 °C). The alternative fibers had contact angles of less than 90°, showing high surface energy. Since the alternative fibers had a low Young's modulus and tensile strength (5.57–8.52 GPa and 57.45–71.26 MPa, respectively), they are suitable for some specific applications in the biomedical industry. In contrast, conventional fibers are suitable where a higher stiffness and strength is required. [ABSTRACT FROM AUTHOR]

Research on the methods of increasing the productivity of a new variety of the white sweet clover, called Almaz, was carried out in 2017-2020 at the experimental field of a separate subdivision of the Federal Center for Bast Fiber Crops in Penza. The application of micro- and biofertilizers (Megamix Profi, GUMI 20, AgroVerm) in pre-sowing seed treatment and foliar fertilizing was studied. The application of mineral fertilizers Р60К60 on formation of the white sweet clover agrocenosis in the year of sowing, nutritive value and productivity in the year of use were also studied. As a result of studies it was found that under the conditions of arid and hyperarid periods of the melilot vegetation in 2017-2020 the studied preparations had a positive effect on the formation of agrocenosis and indicators of plant development by the end of the growing season of the first year of plant life. Thus, the germination and safety of plants, as well as increased values of plant height, number of wintering buds, weight and root volume, respectively. The effect of seed treatment and foliar fertilizing with bio- and microfertilizers was evident in the year of use by an increase in dry matter yield by 4.7-11.1%; the use of mineral fertilizers provided an additional 10.5 % of dry matter yield. The seed treatment was more effective than foliar fertilization, with a 3.8 % higher dry matter yield. An improvement in the nutritive value of the green mass of the sweet clover was noted – the crude protein content increased by 0.3-0.8 %, the crude fiber content decreased by 0.3-0.7 %. The content of exchange energy, feed units in one kg of dry matter and digestible protein in one feed unit varied slightly in the options.

White sweet clover (Melilotus albus) cultivation, despite its many positive features such as the high yield of green mass, as well as protein content, is currently not widespread. This study aimed to determine the effect of the different sowing densities (500, 1000, and 1500 seeds per m2) and harvesting at different growth stages (before budding, budding, and full flowering) on the quality of fodder from the white sweet clover (Adela variety). The field experiment was carried out in the years 2018–2020 in a split-plot design. Plant biometric measurements, green and dry yield, the content of total protein, crude fat, crude fiber, ash, macro and microelements, and coumarin content were determined. The results show that an increase in the sowing density significantly increased the plant population after emergence and green and dry matter. However, it does not affect the total protein, crude fat, crude fiber, ash, macro and microelements, and coumarin content. Harvesting plants at the flowering stage increased plant height and green fodder yield. Plants harvested at the prebudding phase were characterized by the highest share of leaves (40.3%) and were the most abundant in protein (21.7%) and minerals (ash content 12.71%). Unfortunately, sweet clover at this growth stage contained the highest level of coumarin, which limits its use in animal feed. [ABSTRACT FROM AUTHOR]

Summary: Melilotus species are used as green manure and rotation crops worldwide and contain abundant pharmacologically active coumarins. However, there is a paucity of information on its genome and coumarin production and function. Here, we reported a chromosome‐scale assembly of Melilotus albus genome with 1.04 Gb in eight chromosomes, containing 71.42% repetitive elements. Long terminal repeat retrotransposon bursts coincided with declining of population sizes during the Quaternary glaciation. Resequencing of 94 accessions enabled insights into genetic diversity, population structure, and introgression. Melilotus officinalis had relatively larger genetic diversity than that of M. albus. The introgression existed between M. officinalis group and M. albus group, and gene flows was from M. albus to M. officinalis. Selection sweep analysis identified candidate genes associated with flower colour and coumarin biosynthesis. Combining genomics, BSA, transcriptomics, metabolomics, and biochemistry, we identified a β‐glucosidase (BGLU) gene cluster contributing to coumarin biosynthesis. MaBGLU1 function was verified by overexpression in M. albus, heterologous expression in Escherichia coli, and substrate feeding, revealing its role in scopoletin (coumarin derivative) production and showing that nonsynonymous variation drives BGLU enzyme activity divergence in Melilotus. Our work will accelerate the understanding of biologically active coumarins and their biosynthetic pathways, and contribute to genomics‐enabled Melilotus breeding. [ABSTRACT FROM AUTHOR]

In this paper, a possible use of white sweet clover (Melilotus albus Med.) for phytoremediation was assessed. The plants were grown on soils with naturally occurring concentrations of potentially toxic elements (PTEs). First, the content of PTEs was determined in plant biomass and in soil samples using: (a) Optical emission spectrometry with inductively coupled plasma to determine Sb, As, Cd, Cu, Ni, Pb, and Se, and (b) thermal decomposition, amalgamation, and atomic absorption spectrometry to determine Hg. The effectiveness of Melilotus albus Med. (M. albus) for phytoremediation was evaluated using the bioconcentration factor (BCF). The phytoextraction potential of M. albus was determined using bioaccumulation factor (BAC) and translocation factor (TF) values. The highest concentration of PTEs in roots was detected for zinc (10.56 mg/kg of dry weight, DW) and copper (5.128 mg/kg of DW). Similarly, the highest concentration in above-ground parts of the plant was detected for zinc and copper (12.638 and 4.0 mg/kg of DW, respectively). Although the values were relatively high, the effectiveness of the absorption of these PTEs by plant biomass from the soil was relatively very low. BAC and BCF were always lower than 1. On the other hand, the results suggested that M. albus effectively transports PTEs (only for Zn, Pb and Hg) from roots to shoots, because TF was always higher than 1. However, the accumulation of PTEs from soils with a natural abundance of PTEs was not excessive in comparison to conventional maize silage. Therefore, there is no potential risk of biomethane production in biogas plants when biomass from M. albus is used.

Cropping system characteristics such as tillage intensity, crop identity, crop-livestock integration and the application of off-farm synthetic inputs influence weed abundance, plant community composition and crop-weed competition. The resulting plant community, in turn, has species-specific effects on soil microbial communities which can impact the growth and competitive ability of subsequent plants, completing a plant–soil feedback (PSF) loop. Farming systems that minimize the negative impacts of PSFs on subsequent crop growth can increase the sustainability of the farming enterprise. This study sought to assess the individual and combined impact of the cropping system (certified organic-grazed, certified organic till and conventional no-till) and crop sequence [pairwise rotations with safflower (Carthamus tinctorius), yellow sweet clover (Melilotus officinalis) and winter wheat (Triticum aestivum)] on the PSF magnitude and direction. All cropping systems followed the same 5-year rotation and had completed one full rotation before soil was sampled. In a greenhouse setting, a sterile soil mix was inoculated with field soil collected from all systems and three crops. The PSF study consisted of two stages (conditioning and response phases) that mimicked the rotation stages occurring in the field. PSFs were calculated by comparing the biomass of the response phase plants grown in inoculated and uninoculated soils. The farm management system affected PSFs, inferring that tillage reduction can encourage more positive PSFs. Crop sequence did not affect PSF but interacted strongly with the farm system. As such, the effects of the farming system on PSFs are best illustrated when taken into account with the identity of the previous and current crops of a cropping sequence. [ABSTRACT FROM AUTHOR]

White sweet clover (Melilotus albus) cultivation, despite its many positive features such as the high yield of green mass, as well as protein content, is currently not widespread. This study aimed to determine the effect of the different sowing densities (500, 1000, and 1500 seeds per m2) and harvesting at different growth stages (before budding, budding, and full flowering) on the quality of fodder from the white sweet clover (Adela variety). The field experiment was carried out in the years 2018–2020 in a split-plot design. Plant biometric measurements, green and dry yield, the content of total protein, crude fat, crude fiber, ash, macro and microelements, and coumarin content were determined. The results show that an increase in the sowing density significantly increased the plant population after emergence and green and dry matter. However, it does not affect the total protein, crude fat, crude fiber, ash, macro and microelements, and coumarin content. Harvesting plants at the flowering stage increased plant height and green fodder yield. Plants harvested at the prebudding phase were characterized by the highest share of leaves (40.3%) and were the most abundant in protein (21.7%) and minerals (ash content 12.71%). Unfortunately, sweet clover at this growth stage contained the highest level of coumarin, which limits its use in animal feed.

The ever-growing number of biogas plants also leads to an increasing demand for suitable, alternative plant substrates. A currently dominant plant substrate is maize silage. However, intensive cultivation of maize (Zea mays L.) as an energy crop in monocultures poses risk to the environment (soil erosion, depletion of soil nutrient supplies, increased concentration of pests—Ostrinia nubilalis). In this study, results of experimental methane production from silages of alternative substrates, such as fodder mallow (Malva verticillata var. crispa L.; FM), white sweet clover (Melilotus albus Medik.; WSC) and their mixture, are presented. Based on the dry matter yield of a mixed culture of mallow and sweet clover, the value of the land equivalent ratio parameter (LER) was set as 1.05. The obtained LER (>1) value shows that the cultivation of the two tested crops in the mixed culture is more beneficial than their monocultures. Methane production from all three silage variants was fully comparable with results of methane production from the maize silage. Anaerobic fermentation of the mixture of FM and WSC did not result in higher methane yield than the average result of monosubstrates.

Dicoumarol is a mycotoxin, that acts as a blood anticoagulant, is formed during the microbial action of molds and fungi in spoiled hay or silage containing high-coumarin plant. A HPLC-DAD method for determination of coumarins, including dicoumarol, coumarin, and 4-hydroxycoumarin was developed. Methanol and acetic acid were used as mobile phase with gradient elution. The simultaneous separation was performed using C18 type of stationary phase. The recoveries were 88.6 – 92.6 %, 91.8 – 95.0 %, and 89.7 – 94.1 % (evaluated for three concentration levels) for dicoumarol, coumarin, and 4-hydroxycoumarin respectively. The parameters of system suitability (repeatability of retention times and peak areas) were determined for evaluation of the method. The method showed a good linearity in the concentration range 0.7 – 100 μg.mL−1 for dicoumarol, 0.05 – 100 μg.mL−1 for coumarin and 4-hydroxycoumarin with correlation coefficients higher than 0.9885. Extracts of sweet clover herb, hay, and spoiled hay were subjected to HPLC-DAD analysis. The most abundant compound in sweet clover herb and hay extracts was coumarin. In spoiled sweet clover hay extract the 4-hydroxycoumarin was detected in addition. The formation of 4-hydroxycoumarin was also observed in the synchronous fluorescence spectra recorded at the wavelength difference of 90 nm (difference between emission and excitation wavelength).

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Coumarin is a plant secondary metabolite responsible for the health-promoting properties of the Melilotus officinalis herb used in traditional medicine in the treatment of lymphedema and chronic venous diseases. The aim of this study was to analyze the possibility of honey enrichment with coumarin derived from sweet clover (Melilotus) in two different ways: herb honey and herb-infused honey production. Herb honey was obtained by feeding bees with sugar syrup enriched with a M. officinalis herb water infusion, whereas herbal macerates in multifloral honey were prepared with crushed flowers (fresh and dried) of M. officinalis and Melilotus albus at 20 °C in dark place for 6 months. Melilotus albus nectar honey (n = 14) was used as control. Using HPLC analysis, the highest content of coumarin (from 41.33 to 137.89 mg/kg) and its precursor o-coumaric acid (from 6.57 to 25.15 mg/kg) in M. officinalis flowers-infused honey were found. Herb honey contains 4.48 mg/kg coumarin and for the first time, this compound was determined in nectar honey from Polish M. albus honey (0.05 to 0.88 mg/kg). The addition of Melilotus flowers enhanced antioxidant activity (measured by FRAP, DPPH and PCL methods) of infused-honey in comparison to multifloral honey (control) and herb honey. The coumarin and o-coumaric acid were weakly correlated with antioxidant activity. It was found that only herb-infused honey can be proposed as the natural safe remedy for venous diseases prevention. [ABSTRACT FROM AUTHOR]

Crop yields are mostly dependent on the climatic conditions and structural indexes where perennial and annual herbs have special value as previous crops. White melilot is an important annual herb that should be introduced into the crop rotations of the Steppe zone. Seed productivity of white melilot depends on a number of factors including the growth conditions and features of the cultivated variety. The best results in the increase of seed productivity of the crop could be achieved by the optimal agrotechnological and chemical measures of weed management herewith the application of herbicides is characterized with the highest efficiency. The three-year study was conducted in the conditions of the Steppe zone of Ukraine on the dark-chestnut soil to determine the best option for chemical weed management in the crops of white melilot. It was determined that the highest seed productivity of the crop with the average seed yield of 840.0 kg/ha and best sowing qualities of the obtained seeds (the energy of sprouting of 81%, seedling rate of 85%, 1000 seeds weight of 2.06 g) were achieved on the variant with application of herbicide Pulsar 40 (active substance imazamox, 40 g/l) in the dose of 1.0 l/ha. Therefore, we recommend the utilization of imazamox-based herbicide for weed management in the crops of white melilot under its cultivation in the conditions of the Steppe zone. [ABSTRACT FROM AUTHOR]

Background and Aims Submergence is a severe stress for most plants. Melilotus siculus is a waterlogging- (i.e. root zone hypoxia) tolerant annual forage legume, but data were lacking for the effects of partial and full submergence of the shoots. The aim was to compare the tolerance to partial and full submergence of 15 M. siculus accessions and to assess variation in traits possibly contributing to tolerance. Recovery ability post-submergence was also evaluated. Methods A factorial experiment imposed treatments of water level [aerated root zone with shoots in air as controls, stagnant root zone with shoots in air, stagnant root zone with partial (75 %) or full shoot submergence] on 15 accessions, for 7 d on 4-week-old plants in a 20/15 °C day/night phytotron. Measurements included: shoot and root growth, hyponastic petiole responses, petiole gas-filled spaces, leaflet sugars, leaflet surface hydrophobicity, leaflet gas film thickness and phellem area near the base of the main root. Recovery following full submergence was also assessed. Key Results Accessions differed in shoot and root growth during partial and full shoot submergence. Traits differing among accessions and associated with tolerance were leaflet gas film thickness upon submergence, gas-filled spaces in petioles and phellem tissue area near the base of the main root. All accessions were able to re-orientate petioles towards the vertical under both partial and full submergence. Petiole extension rates were maintained during partial submergence, but decreased during full submergence. Leaflet sugars accumulated during partial submergence, but were depleted during full submergence. Growth resumption after full submergence differed among accessions and was positively correlated with the number of green leaves retained at desubmergence. Conclusions Melilotus siculus is able to tolerate partial and full submergence of at least 7 d. Leaflet surface hydrophobicity and associated gas film retention, petiole gas-filled porosity and root phellem abundance are important traits contributing to tolerance. Post-submergence recovery growth differs among accessions. The ability to retain green leaves is essential to succeed during recovery. [ABSTRACT FROM AUTHOR]

Abstract Invasive species control requires understanding the mechanisms behind their establishment and their interactions with other species. One potential ecosystem alteration influencing the establishment and spread of invasive species is anthropogenic nitrogen enrichment, from sources like introduced or invasive nitrogen (N)-fixing legumes, which can alter competition between native, non-native, and invasive plants. Kentucky bluegrass (Poa pratensis) and N-fixing yellow sweet clover (Melilotus officinalis) are exotic to the Great Plains and are currently invading and degrading native rangelands by altering ecosystem processes and displacing native plants. Therefore, we investigated how N enrichment from yellow sweet clover affects the aboveground biomass production of Kentucky bluegrass and western wheatgrass (Pascopyrum smithii), a native cool-season grass, the ranges of which overlap in the northern Great Plains. In a controlled greenhouse environment, we conditioned experimental pots by growing yellow sweet clover and terminating each plant after 8 wk. Conditioned soils contained ≈ 340% more plant-available N than untreated soils 2 wk after yellow sweet clover death. We then grew Kentucky bluegrass and western wheatgrass transplant seedlings in interspecific and intraspecific pairs in pots conditioned either with or without yellow sweet clover for 12 wk. Aboveground biomass production of both Kentucky bluegrass and western wheatgrass grown in interspecific and intraspecific pairs increased in conditioned soils. However, when grown together in conditioned pots, the increase in Kentucky bluegrass biomass relative to untreated pots (520%) was double that of the increase in western wheatgrass biomass (260%). Our results reveal that Kentucky bluegrass can use increased soil N to produce proportionally more aboveground biomass than western wheatgrass, a native grass competitor. Thus, our results suggest yellow sweet clover and other sources of N enrichment may facilitate the invasion of Kentucky bluegrass. [ABSTRACT FROM AUTHOR]

To improve the serious situation of soil degradation in China, tests were carried out to study the improvement effects of Russian Melilotus officinalis on the low base saturation acid soil, the alkaline-saline soil and the heavy metal contaminated soil. The results showed that M. officinalis could improve the low base saturation acid soil, increase the salt, alkali content, mineral nutrition and organic nutrient, which indicated that M. officinalis could be planted in the acid soil, and had a significant ecological eefect on soil improvement. M. officinalis achieved a remarkable eefect on improving the alkaline-saline soil (pH 8.5 - 9.5) that the soil desalted, dealkalized and had more organic nutrient. Moreover, M. ofifcinalis could improve the heavy metal contaminated soil and had the ability to enrich and transform heavy metal elements, especially Cd and Ni, thereby presenting significant repair eefects on soil improvement. Therefore, Russian M. ofifcinalis had multiple--fects on degraded soil improvement, so it is worthy of application and popularization. [ABSTRACT FROM AUTHOR]

The work presents results of studies conducted from 2008 to 2011 to determine the effect of undersown catch crops, which were either autumn-incorporated or left on the soil surface as mulch for spring incorporation, on the content microelements in potato tubers cultivated in various production systems. The following two factors were examined: I. - manuring with an undersown catch crop: control, farmyard manure, white melilot, white melilot + westerwolds ryegrass, westerwolds ryegrass, white melilot applied as mulch, white melilot + westerwolds ryegrass applied as mulch, westerwolds ryegrass applied as mulch; II. - production system: integrated and organic. Potato tuber samples were taken to determine microelements. The results demonstrated that manuring of potato with undersown catch crops, in particular autumnincorporated white melilot, increased iron, zinc and boron contents, and reduced copper and manganese contents in potato tubers. Potatoes cultivated in the integrated production system had a higher concentration of copper, manganese and zinc whereas organic tubers had more iron and boron. [ABSTRACT FROM AUTHOR]

The sensory-chemical profiles of Finnish honeys (labeled as buckwheat, cloudberry-bog, lingonberry, sweet clover, willowherb and multifloral honeys) were investigated using a multi-analytical approach. The sensory test (untrained panel, n  = 62) was based on scaling and check-all-that-apply (CATA) methods accompanied with questions on preference and usage of honey. The results were correlated with corresponding profiles of odor-active compounds, determined using gas chromatography coupled with mass spectrometry/olfactometry (GC–MS/O). Botanical origins and chemical compositions including sugars were evaluated using NMR spectroscopy. A total of 73 odor-active compounds were listed based on GC–O. Sweet and mild honeys with familiar sensory properties were preferred by the panelists (PCA, R 2 X (1) = 0.7) while buckwheat and cloudberry-bog honeys with strong odor, flavor and color were regarded as unfamiliar and unpleasant. The data will give the honey industry novel information on honey properties in relation to the botanical origin, and consumer preference. [ABSTRACT FROM AUTHOR]

Melilotus is an important genus of legume plants and an herbage with excellent nitrogen fixation; it can tolerate extreme environmental conditions and possesses important medicinal value. However, there is limited genetic information about the genus; thus, we analysed four chloroplast loci (rbcL, matK, psbA-trnH and trnL-F) and one nuclear region (ITS) to determine the genetic diversity of 199 accessions from 18 Melilotus species. The rbcL and matK sequences were highly conserved, whereas the trnL-F and ITS sequences contained variable and parsimony-informative sites. In our analyses of the single and combined regions, we calculated the pairwise distance, haplotype and nucleotide diversity and gaps and then constructed phylogenetic trees to assess the genetic diversity, and our results revealed significant variations among the different accessions. The genetic distance values were between zero and nine, and based on the combined regions, the highest frequency value was approximately four. Melilotus showed high haplotype and nucleotide diversity, particularly in the ITS sequences, with values of 0.86 and 0.0087, respectively. The single ITS sequence, psbA-trnH, and the combined matK+rbcL+trnL-F (MRT) and matK+rbcL+psbA-trnH+trnL-F+ITS (MRPTI) regions showed interspecific variation in the gap analysis. Phylogenetic trees calculated using ITS, psbA-trnH and MRPTI sequences indicated distinct genetic relationship in 18 species, and these species could be divided into two groups. By determining the genetic diversity of plants, we can evaluate the genetic relationships among species and accessions, providing a basis for preserving and utilizing the genetic resources of Melilotus. [ABSTRACT FROM AUTHOR]

Background and Aims Soil waterlogging adversely impacts most plants. Melilotus siculus is a waterloggingtolerant annual forage legume, but data were lacking for the effects of root-zone hypoxia on nodulated plants reliant on N2 fixation. The aim was to compare the waterlogging tolerance and physiology of M. siculus reliant on N2 fixation or with access to NO3-. Methods A factorial experiment imposed treatments of water level (drained or waterlogged), rhizobia (nil or inoculated) and mineral N supply (nil or 11 mm NO3-) for 21 d on plants in pots of vermiculite in a glasshouse. Nodulation, shoot and root growth and tissue N were determined. Porosity (gas volume per unit tissue volume) and respiration rates of root tissues and nodules, and O2 microelectrode profiling across nodules, were measured in a second experiment. Key Results Plants inoculated with the appropriate rhizobia, Ensifer (syn. Sinorhizobium) medicae, formed nodules. Nodulated plants grew as well as plants fed NO3-, both in drained and waterlogged conditions. The growth and total N content of nodulated plants (without any NO3- supplied) indicated N2 fixation. Respiration rates (mass basis) were highest in nodules and root tips and lowest in basal root tissues. Secondary aerenchyma (phellem) formed along basal root parts and a thin layer of this porous tissue also covered nodules, which together enhanced gas-phase diffusion of O2 to the nodules; O2 was below detection within the infected zone of the nodule interior. Conclusions Melilotus siculus reliant on N2 fixation grew well both in drained and waterlogged conditions, and had similar tissue N concentrations. In waterlogged conditions the relatively high respiration rates of nodules must rely on O2 movement via the aerenchymatous phellem in hypocotyl, roots and the outer tissue layers of nodules. [ABSTRACT FROM AUTHOR]

This paper explores the salt-tolerance response ofMelilotusspecies, a forage legume, based on tolerance/susceptibility indices, leaf water relations, leaf/root ion concentrations and multivariate (factor and function discriminate) analyses. In a greenhouse experiment, salinity-susceptibility of 12 accessions ofMelilotus(M. albus, M. indicus, M. officinalis) in addition, two controls (alfalfa and Persian clover) were evaluated at 0 mM, 100 mM, 200 mM, and 300 mM of sodium chloride (NaCl). The experiment was conducted in a split plot design with two replications. Stress tolerance score (STS) was calculated based on tolerance/susceptibility indices. Salt-tolerant accessions were identified. An extracted bi-plot based on factor analysis confirmed the results of tolerance/susceptibility indices. There were significant correlations between STS and multivariate statistics. Root potassium cation (K+) concentration was higher inMelilotusspecies than the control species in all the salinity treatments. STS, root K+concentration, leaf turgidity, and leaf succulence characteristics can be used for screening salinity-tolerance inMelilotusaccessions. In all treatments,M. officinalis, M. indicus, andM. albuswere salinity-tolerant species in descending order. Abbreviations: Total biomass yield under control (Yp) and stress (Ys) mediums, geometric mean productivity (GMP), leaf area (LA), leaf tissue density (LTD), linear regression coefficient (b), mean productivity (MP), relative water content (RWC%), Succulence (S), specific leaf area (SLA), stress susceptibility index (SSI), stress tolerance index (STI), tolerance (TOL), water content at saturation (WCS), water saturation deficit (WSD), sodium chloride (NaCl), potassium cation (K+), sodium cation (Na+), analysis of variance (ANOVA), least significant difference (LSD), alfalfa control accession (CHA), Persian clover control accession (CHC), coefficient of variation (CV), statistics of factor (F) and discriminate function (D) [ABSTRACT FROM AUTHOR]

The prospects of creating adaptive self-regulating herbage mixtures and their longevity in the context of different methods of use were studied at the sites with cryogenic pale-yellow soil in Yakutia (Sakha Republic). We have used a six-component grass-legume mixture. Research results have showed that grass-legume mixtures intended for grazing provide high yields of feed crops (7.75 tons per year on average) at late sowing dates and the cutting height of 5 cm. The share of alfalfa in herbage mixtures is 61%. In pure batches of alfalfa and sweet clover, the maximum content of crude protein was 23.89% and 23.5%, fat content - 3.86% and 3.38%, respectively. In the pasture coenosis, organic matter content is the highest (up to 4.37%) at the cutting depth of 0-10 cm and 10-20 cm regardless the sowing period. [ABSTRACT FROM AUTHOR]

Melilotus albus Medik. is a legume species with potential to be used as a forage or conserved fodder in Northwest China. Breeding M. albus as a forage crop is focused on simultaneously improving forage yield and reducing coumarin (Cou) content. The goal of the present study was to estimate genotypic variation for several key agronomic traits and to evaluate the impact of single-trait selection and Smith--Hazel-index-based multitrait selection for herbage dry weight (DW), plant height (PH), stem number (SN), and leaf area (LA). A quantitative genetic analysis was conducted on trait measurements from 25 half-sib (HS) families evaluated at Yuzhong and Linze, in China. Significant (P < 0.05) genotypic variation among the HS families for all traits was observed. There was also significant (P < 0.05) genotype x environment interaction for all traits except for LA. The predicted genetic gain, based on estimated genotypic variance, for single-trait selection varied from 2.3 to 30.3% per selection cycle. Pattern analysis of the HS family ´ multitrait best linear unbiased prediction matrix provided a graphical summary of the association among the traits, as well as the four HS family groups generated from clustering. Some members within Group 1 could be crossed to generate a breeding population to be used in recurrent selection for M. albus cultivar development in Northwest China. The applicability of a Smith--Hazel selection index to increase expression of the traits DW, PH, and SN and reduce LA, indirectly reducing Cou content, was confirmed. This index enabled identification of families for increasing key biomass traits and decreasing Cou production. [ABSTRACT FROM AUTHOR]

Melilotus, an annual or biennial herb, belongs to the tribe Trifolieae (Leguminosae) and consists of 19 species. As an important green manure crop, diverse Melilotus species have different values as feed and medicine. To identify different Melilotus species, we examined the efficiency of five candidate regions as barcodes, including the internal transcribed spacer (ITS) and two chloroplast loci, rbcL and matK, and two non-coding loci, trnH-psbA and trnL-F. In total, 198 individuals from 98 accessions representing 18 Melilotus species were sequenced for these five potential barcodes. Based on inter-specific divergence, we analysed sequences and confirmed that each candidate barcode was able to identify some of the 18 species. The resolution of a single barcode and its combinations ranged from 33.33% to 88.89%. Analysis of pairwise distances showed that matK+rbcL+trnL-F+trnH-psbA+ITS (MRTPI) had the greatest value and rbcL the least. Barcode gap values and similarity value analyses confirmed these trends. The results indicated that an ITS region, successfully identifying 13 of 18 species, was the most appropriate single barcode and that the combination of all five potential barcodes identified 16 of the 18 species. We conclude that MRTPI is the most effective tool for Melilotus species identification. Taking full advantage of the barcode system, a clear taxonomic relationship can be applied to identify Melilotus species and enhance their practical production. [ABSTRACT FROM AUTHOR]

Most of Melilotus Miller species, a forage legume, contain coumarin, an unfavorable component in their shoot. There is no information concerning coumarin on Iranian Melilotus germplasms. It is crucial for plant breeders to know the concentration of this component in the breeding materials, its relationship with other traits and with genotype geographic origin. In a two-year field study a total of 196 accessions of biennial Melilotus were planted in two replications in March 2014. Thirty-nine agro-morphobiochemical traits were recorded according to standard descriptors. Fluorescencemetery assay was applied to detect free, conjugated, and total coumarin concentrations in plant leaves. The data were subjected to correlation, path, regression, factor, and cluster analyses. Total coumarin concentration varied from 0.09 to 5.27% of leaf dry weight (LDW) with a mean of 1.63 ± 0.08% of LDW among accessions. In M. officinalis, correlation analysis showed no significant correlation between coumarin concentration and cotyledon, hypocotyl, and stem colors. Therefore, no early predicting trait was determined to estimate coumarin concentration in the early growth phases of the germplasms. A positive linear relationship between leaf area and total coumarin concentration was observed by path and regression analyses. Based on factor analysis and extracted bi-plot, the scatter of accessions with low coumarin and high forage yield was determined. Materials were divided into three groups by k-means cluster analysis. There was no clear pattern between coumarin concentration and genotype geographic origin. The findings could be utilized by Melilotus breeders to apply in their breeding programs. [ABSTRACT FROM AUTHOR]

This study aimed to measure the effects of elevated levels of Na + and Cl − on ion concentration, growth, and photosynthetic performance of Melilotus siculus , Tecticornia pergranulata , and Thinopyrum ponticum in pot trials under controlled glasshouse conditions. Experiments were conducted using Na + and Cl − dominant saline soil treated with constituted saline solutions (0, 2.0 and 4.0 dS m −1 Na + or Cl − ). Maximum Na + and Cl − concentration was in T. pergranulata shoots; maximum Na + concentration was in T. pergranulata roots, whereas maximum Cl − in T. ponticum roots. Maximum quantum yield of PSII [Fv/Fm] values did not change significantly. Net-photosynthetic rate [ P n] and stomatal conductance [g s ] values increased and non-photochemical quenching [NPQ] value decreased significantly in T. pergranulata with increasing Na + dominant treatments and the reverse occurred in Cl − dominant treatments. Pn, g s , and NPQ values did not significantly change in T. ponticum with increasing Na + and Cl − dominant treatments. The shoot and root biomasses of T. pergranulata were the maximum at 4.0 dS m −1 Na + dominant treatment. In T. ponticum , the maximum shoot and root fresh masses and shoot dry mass occurred in the control treatment and the maximum root dry mass occurred in the 2.0 dS m −1 Cl − dominant treatment. In T. pergranulata , strong positive correlations occurred between Na + concentration in shoots and salt tolerance (shoot mass over control) and moderately positive correlation occurred between Cl − concentration in shoots and salt tolerance. However, strong negative correlations were observed between Na + and Cl − concentration in shoots and salt tolerance in T. ponticum as well as between Cl − concentration in shoots and salt tolerance in M. siculus . Our glasshouse experiments indicate that 4.0 dS m −1 Cl − solutions in the soil was more intensely damaging than that of Na + . The greatest concentration of both Na + and Cl − was in T. pergranulata under the tested circumstances indicating it to be the candidate of choice for restoring saline soils, followed in capacity by M. siculus and T. ponticum . [ABSTRACT FROM AUTHOR]

A case study of Polish Melilotus officinalis honey was presented for the first time. Gas chromatography–mass spectrometry (GC-MS) (after steam distillation, Soxhlet extraction, ultrasonic solvent extraction, and solid phase extraction (SPE)) and targeted high performance liquid chromatography with a photodiode array detector (HPLC-PAD) were applied to determine the characteristic components of honey. While ubiquitous in most honeys, carbohydrates, terpene derivatives, and phenylacetic acid dominated in the Soxhlet extracts (25.54%) and in the application of SPE (13.04%). In addition, lumichrome (1.85%) was found, and may be considered as a marker of this honey. Due to the presence of these compounds, Polish yellow sweet clover honey is similar to French lavender honeys. The major compounds determined in the methanolic extract were (+)-catechine (39.7%) and gallic acid (up to 30%), which can be regarded as specific chemical markers of the botanical origin of melilot honey. With respect to total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were determined spectrophotometrically. The honey exhibited a moderate antioxidant activity, typical for light honeys, which correlates well with its phenolic and flavonoid composition. [ABSTRACT FROM AUTHOR]

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Allodynia is a main symptom of the peripheral neuropathy induced by vincristine treatment. Theoesberiven F, a combination of Melilotus extract and proxyphylline, is currently used for the treatment of inflammatory conditions due to its potent anti-inflammatory and analgesic properties. The anti-allodynic effect of theoesberiven F on mechanical and cold allodynia in a rat model of vincristine-induced neuropathy was investigated in the present study. Intraperitoneal vincristine injections were administered to male Sprague-Dawley rats at a dose of 0.1 mg/kg/day over 12 days (5 days of injection, a 2-day cessation and 5 days of injection). Rats that were allodynic following the vincristine injections were randomly allocated into four groups. Normal saline was injected into rats in the control group. Theoesberiven F (0.1, 0.25 and 0.5 mg/kg) was administered to rats in the three experimental groups, respectively. Mechanical and cold allodynia were measured at preadministration and at 15, 30, 60, 90, 120, 150 and 180 min following the intraperitoneal administration of normal saline or theoesberiven F. The decreased paw withdrawal threshold induced by vincristine injection was increased by theoesberiven F administration. The increased withdrawal frequency to cold stimuli developed by vincristine was reduced by theoesberiven F administration. The results of this study suggest that the administration of theoesberiven F may be beneficial in reducing the mechanical and cold allodynia developed during vincristine treatment. [ABSTRACT FROM AUTHOR]

Sweet clovers are legumes able to grow on most soils, and two sweet clover species,Melilotus albusandMelilotus officinalishave been introduced and are now cultivated on estuary land. We characterized the composition and morphology of sweet clover stems collected after the seeds had reached maturity. We also carried out histochemical analyses on transverse sections. The two species had similar morphological structures, which two fiber fractions: flexible long fibers and stiff, dense shives, accounting for about 12% and 88% of stem dry matter, respectively. Histological analysis revealed the presence of bundles of highly cellulosic bast fibers (lignocellulosic material: 71–78% of dry matter). The shives are a natural mesoporous material composed of 85–90% lignocellulosic fibers. Both fiber fractions displayed good thermal resistance to temperatures up to 225°C and a moderate affinity for water. These two types of fibers are similar to those of flax and hemp, suggesting their possible use for the same types of applications. Sweet clovers therefore constitute a new source of fibers that can be cultivated on wet and saline soils not otherwise suitable for agriculture. [ABSTRACT FROM AUTHOR]

Melilotus is one of the most important legume plants, in part due to its production of coumarin, but the genetic diversity among the 18 species remains unclear. In the present study, the analysis of genetic diversity of the Melilotus species was performed with simple sequence repeat (SSR) markers. From the PCR amplification, we selected 18 out of 70 primers that were used in previous studies. Three hundred twenty-six sampled individuals were assayed to study the genetic diversity and polymorphisms based on the SSR markers. All analyzed markers were polymorphic, and 287 alleles were identified, with 15.94 alleles per locus detected. The polymorphism information content ( PIC ) values ranged from 0.71 to 0.93, with an average of 0.87, which indicates that the markers were highly informative. Based on the unweighted pair-group method with arithmetic mean (UPGMA) cluster analysis, we found that the 18 species were divided into two clusters. M. italicus and M.​ speciosus from cluster A and M. indicus and M. segetalis from cluster B were closely related. Population structure analysis suggested that the optimum number of groups was three. From the analysis of molecular variance, 17.79% of the variance was due to species differences, 31.61% of the variance was due to differences among populations within species and the remaining 50.60% was due to differences within populations. The results of the present study showed that these SSR markers will benefit the Melilotus research community for genetics and breeding. Furthermore, this study also established the foundation for future breeding programs, genetic improvement and coumarin production in the Melilotus species. [ABSTRACT FROM AUTHOR]

Background: Melilotus officinalis Linn. (Fabaceae) historically has been used for a variety of medicinal purposes. Despite the popular medicinal utilization, still no conclusive study has been reported so far regarding the pharmacognostical standardization. Aim: Thus, the present study was focused to scientifically establish a standard monograph of M. officinalis on the basis of pharmacognostical parameters. Material and methods: The detailed macroscopic and qualitative as well as quantitative microscopic characters of M. officinalis were analyzed. Results: The morphological characters of M. officinalis were established. The transverse section of leaf shows lamina with single layered epidermis, anomocytic stomata, mesophyll, midrib with single layered epidermis and vascular strands were present in lamina. Stem contains single layered epidermis, cortex with several layers, phloem, xylem and starch grains. Root shows the presence of cork, phloem, xylem and starch grains. The powder characteristics of M. officinalis revels the presence of Epidermis, stomata, cork, parenchyma, mesophyll, fibers, spiral and pitted xylem vessels. The quantitative microscopy of leaf of M. officinalis reveals Stomatal number Upper surface (28.75 ± 2.5), Lower surface (38.37 ± 2.5); Stomatal index Upper surface (13.10 ± 1.50) Lower surface (12.58 ± 1.10); Vein islet number (96.25 ± 4.79), Vein termination number (27.50 ± 2.87) and Palisade ratio 6.5 ± 0.58. Conclusion: The obtained qualitative and quantitative standards would serve as a useful guide for correct identification, purity, standardization and preparation of monograph of M. officinalis. [ABSTRACT FROM AUTHOR]

The establishment of abundantly flowered, highly rewarding non-native plant species is expected to have strong consequences for native plants through altered pollination services, particularly in boreal forest where the flowering season is short and the pollinator pool is small. In 18 boreal forest sites, we added flowering Melilotus albus to some sites and left some sites as controls in 2 different years to test if the invasive plant influences the pollination and reproductive success of two co-flowering ericaceous species: Vaccinium vitis- idaea and Rhododendron groenlandicum. We found that M. albus increased the pollinator diversity and tended to increase visitation rates to the focal native plant species compared to control sites. Melilotus albus facilitated greater seed production per berry in V. vitis- idaea when we added 120 plants compared to when we added 40 plants or in control sites. In R. groenlandicum, increasing numbers of M. albus inflorescences lowered conspecific pollen loads and percentage of flowers pollinated; however, no differences in fruit set were detected. The number of M. albus inflorescences had greater importance in explaining R. groenlandicum pollination compared to other environmental variables such as weather and number of native flowers, and had greater importance in lower quality black spruce sites than in mixed deciduous and white spruce sites for explaining the percentage of V. vitis- idaea flowers pollinated. Our data suggest that the identity of new pollinators attracted to the invaded sites, degree of shared pollinators between invasive and native species, and variation in resource limitation among sites are likely determining factors in the reproductive responses of boreal native plants in the presence of an invasive. [ABSTRACT FROM AUTHOR]

Melilotus comprises 19 species, while the phylogenetic relationships between species remain unclear. In the present work, three chloroplast genes, rbcL, matK, trnL-F, and one nuclear region, ITS (internal transcribed spacer) belonging to 48 populations of 18 species of Melilotus were sequenced and phylogenetic trees were constructed to study their interspecific relationships. Based on the phylogenetic tree generated in this study using rbcL analysis, the Melilotus genus is clearly monophyletic in the legume family. Both Bayesian and maximum-parsimony approaches were used to analyze the data. The nrDNA ITS provided more informative characteristics (9.8%) than cpDNA (3.0%). Melilotus contains two closely related groups, clade I and clade II. M. spicatus, M. indicus and M. segetalis have a close relationship. M. infestus, M. siculus and M. sulcatus are closely related. The comparing between molecular phylogeny and flower color classification in Melilotus showed that the flower color is not much informative for phylogenetics of this genus. [ABSTRACT FROM AUTHOR]