• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.1
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• pp.7-11
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• 2004

Alfalfa (Medicago sativa L.) plants have been reported to be autotoxic as well as allelopathic. Laboratory and greenhouse experiments through petri-dish and pot test were conducted to determine autotoxic effects of alfalfa leaf and soil extracts on the germination or early seedling growth of alfalfa, and to evaluate allelopathic effects of alfalfa leaf residues on alfalfa, barnyard grass, com, eclipta and soybean. Alfalfa seed germination was delayed depending on aqueous extract concentration, with no difference in final germination after 48 hours. Alfalfa root length was more sensitive to the autotoxic chemicals from leaf extracts than was germination or shoot length. Root growth of alfalfa was significantly inhibited at extract concentration of more than 1 g dry tissue/L (g $\textrm{L}^{-1}$). Hypocotyl growth, however, was not affected by all the concentrations of leaf extracts. Soil extracts from 4-yr-old alfalfa stand significantly reduced alfalfa root length by 66%, while soil extracts from 0,1, and 3yr-old stand stimulated root length up to 14-32% over the control. Residue incorporation with dry matters of alfalfa leaf at 100 g $\textrm{kg}^{-1}$ reduced seedling length of several crop and weed species, ranging from 53 to 87% inhibition. Addition of nutrient solution into alfalfa leaf extracts alleviated alfalfa autotoxic effect. This result indicates alfalfa leaf and soil extracts or residues could exert autotoxic as well as allelopathic substances into soil environments during and after establishment.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.3
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• pp.295-305
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• 1994

Greenhouse and laboratory studies were conducted to investigate the allelopathic potential of alfalfa and vetch residues on soybean and corn using various extract concentrations (0, 5, 10, 15 and 20%, w/v) and residue rates (0, 0.25, 0.5, 0.75 and 1%, w/w). Aqueous extracts of alfalfa (Medicago sativa L.) and vetch (Vicia spp.) exhibited an allelopathic effect on soybean and corn seed germination, seedling length and weight. The degree of inhibition significantly increased as the aqueous extract concentration increased. Alfalfa and vetch 20% extracts reduced soybean seed germination, seedling length and weight by 35%, 57%, 32% and 15%, 42%, 25% respectively, when compared to control. Corn germination, seedling length and weight was inhibited by 20%, 23%, 38% by alfalfa and 19%, 18%, 35% by 20% vetch extracts. Alfalfa and vetch extracts inhibited secondary root formation and branching as the extract concentration increased. Alfalfa and vetch 20% extracts inhibited by 41% and 32% secondary root numbers, respectively as compared to control. It was found that the aqueous extract of alfalfa resulted in greater reduction in germination, seedling length and weight of soybean than that of vetch. Alfalfa and vetch 1% residue rate inhibited soybean plant height by 30% and 10%, leaf area by 31% and 23%, and dry weight by 18% and 1%, nodule number by 27% and 20% also. Alfalfa and vetch residue significantly enhanced plant height, leaf area and dry weight of corn. The maximum stimulation occurred with 0.25% and 1% of alfalfa and vetch residue rates, respectively. Plant height, leaf area, and dry weight increased by 23%, 59%, 58% and 17%, 52%, 94% with alfalfa and vetch residues of 0.25% and 1%, respectively. This study demonstrates that there is an allelopathic potential resulting from alfalfa and vetch residues on soybean growth and yield. It also suggests that these residues may affect crop growth and development due to the inhibitory or stimulatory effects of allelochemicals existing in the residue.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.1
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• pp.30-35
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• 2004

Compositae plants are known to contain biologically active substances that are allelopathic to agricultural crops as well as weed species. Aqueous extracts from leaves of Xanthium occidentale were assayed against alfalfa (Medicago sativa) to determine their allelopathic effects, and the result showed that the extracts applied onto filter paper significantly inhibited seed germination as well as root growth of alfalfa. Untreated seeds germinated in 60h, but extract concentrations greater than 30g $\textrm{L}^{-1}$ delayed seed germination. The extracts significantly inhibited seed germination of alfalfa, and $\beta$-amylase activity of alfalfa and barley seeds during 24-36 hours after treatment. Aqueous extracts of 40 g $\textrm{L}^{-1}$ from X. occidentale were completely inhibited the hypocotyl and root growth of alfalfa. Aqueous leaf extracts showed the highest inhibitory effect and followed by root and stem extracts. Early seedling growth of both alfalfa and barnyard grass (Echinochloa crus-galli) was significantly reduced by methanol extracts. By means of high-performance liquid chromatography, chlorogenic acid and trans-cinnamic acid were quantified as the highest amounts from water and EtOAc fractions, respectively. BuOH and EtOAc fractions of X. occidentale reduced alfalfa root growth more than did hexane and water fractions. The findings of the bioassays for aqueous or methanol extracts reflected that the inhibitory effect of extract was closely related to the level of responsible allelochemicals found in plant extracts.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.5
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• pp.310-315
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• 2000

Alfalfa (Medicago sativa L.) plants have been reported to contain water-soluble substances that are autotoxic as well as allelopathic. Laboratory experiment through a petri-dish assay with imbibed seeds was conducted to evaluate both autotoxic and allelopathic effects of alfalfa leaf extracts on the germination and early seedling growth of alfalfa, red clover, crested wheatgrass, and Russian wildrye. Alfalfa seed germination was delayed dependent on extract concentration, with no difference in final germination at 72 hours. Root growth of alfalfa was stimulated up to 14% above control at very low concentrations of both leaf and stem extracts of alfalfa and was significantly reduced at extract concentration of more than 0.5g dry tissue/L (${gL}_{-1}$). Leaf extracts were generally more autotoxic for root growth than were stem extracts. Hypocotyl growth was not affected by all the concentrations of both leaf and stem extracts. Root length of legumes was more sensitive to the autotoxic chemicals from leaf extracts than was germination or shoot length. Hypocotyl growth of two legume plants and plant height of two grasses were not influenced by extracts. Seed germination and root growth of legumes were more inhibited by aqueous extracts of alfalfa leaf than were those of grasses. This result indicates autotoxic effect of alfalfa leaf extracts seems to be greater than allelopathic effect.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.3
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• pp.285-294
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• 1994

Dried top and root extracts of seven different weed species, fresh top and root extracts, and various concentrations of extract (0, 5, 10 and 15%, W/V) and residue rate (0.0, 0.25. 0.5, 0.75 and 1.0%) of velvetleaf (Abutilon theophrasti Medic.) were used to study their allelopathic effects on alfalfa in the laboratory and greenhouse. Top and root aqueous extracts of common lambsquarter (Chenopodium album L.), giant foxtail (Setaria faberii Herrm), redroot pig-weed (Amaranthus retroflexus L), velvetleaf, large crabgrass (Digitaria sanguinalis L), canada thistle (Cirsium arvense L.) and prostrate knotweed (Polygonium aviculare L.) significantly inhibited germination, seedling length, weight, and vigor in alfalfa. Top growth extracts of weeds exhibited greater allelopathic effects than root extracts. Alfalfa test species, WL-320, responded significantly different to the various weed species extracts in terms of allelopathic effect. The regression slopes of various top extracts showed significant variation with respect to germination percentage. Velvetleaf (b=3.69) extracts were the most inhibitory, while large crabgrass (b=2.39) extracts had the least allelopathic effect on alfalfa. When compared the activity of fresh velvetleaf extract to that of dry velvetleaf extract, dry extract was more inhibitory to alfalfa germination and seedling growth. Germination, seedling length and weight of alfalfa were inversely proportional to the concentration of dried velvetleaf extracts. Seedling emergence and survival percentage was inhibited by velvetleaf residue mixture treatment. Also, more of the toxic effects were observed from the dried top extracts, as compared to extracts from fresh top and root. These results demonstrate the allelopathic activity of different weed species extracts and suggest that weeds may affect crop growth and development due to the inhibitory effects of allelochemicals present in weed tissue.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.5
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• pp.619-628
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• 1995

Crude aqueous extracts from dried leaves, stems, roots, and flowers from both field grown and greenhouse grown alfalfa plants inhibited alfalfa seed germination and seedling growth. The degree of inhibition was greater in the field grown plant extracts. Flowers extract of field grown plant most inhibited alfalfa germination and seedling growth. In the concentration study, the highest concentration of extract (9.0%, w/v) significantly inhibited total alfalfa seed germination by 50% as compared to control. In partitioning study using pot hydroponic culture of plant biomass into leaves, stems, root, LAR products of LWR and SLA exhibited significant variation among four species. This result support that the inhibitory effect of autotoxic substances presenting in alfalfa tissue may be possible interference with the patitioning of biomass into leaf component relative to the total biomass produced by the alfalfa plant. Toxicity of extract was not reduced by adding activated charcoal, Dowex-50W, amberlite to the extract. Toxic substances existing in most plant tissues but mainly above ground foliage are water soluble and stable and may persist in old alfalfa fields. Thus, it is recommended to remove as much as possible of the above growth parts, especially vegetative stage, before one tries to re-establish alfalfa in former field of alfalfa.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.1
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• pp.123-128
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• 2012

This study was conducted to investigate the quality characteristics of tofu (soybean curd) prepared with added alfalfa extracts (Medicago sativa L.). The moisture content, yield, and pH of tofu decreased according to the increase of an added quantity of alfalfa extract, whereas the crude protein and turbidity increased. The tofu made with additional alfalfa extracts gave a similar amount of crude ash with that of the control. The Hunter's L (lightness) color value decreased as the content of alfalfa extract increased, but the a (redness) and b (yellowness) values increased with increasing alfalfa extract concentration. Textural properties for hardness were higher but cohesiveness, springiness, gumminess, and brittleness were lower in alfalfa extracts with added tofu compared to those of control. Isoflavones such as daidzein and genistein content were significantly increased with increasing concentration of alfalfa extracts. A sensory evaluation indicated that the group with a 0.5% alfalfa extract showed the best color, flavor, taste, and overall acceptability results. In conclusion, adding alfalfa extracts could improve the physicochemical and sensory characteristics of tofu.

• Korean Journal of Environmental Agriculture
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• v.18 no.2
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• pp.185-190
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• 1999

Autotoxic substance(s) from alfalfa(Medicago sativa L.) plants reduces germination and growth of adjacent new alfalfa after alfalfa. The autotoxic chemical(s) in alfalfa are clearly unknown. Our objective was to improve the sensitivity of an alfalfa seedling bioassay for evaluating autotoxic leaf extracts. We determined critical extract concentrations that inhibit seed germination and seedling growth, compared two different culture media, and evaluated the effects of extract sterilization on the sensitivity of the assay, by using streptomycin and autoclaving method. An agar medium in petri plate gave better responses of germination and seedling growth to the extracts than using filter paper in the plate. On agar medium, the concentration of extract required to reach 50% inhibition of root length was 2.7 g $kg^{-1}$, and of germination and hypocotyl length were 3.8 and 9.9 g $kg^{-1}$, respectively. Leaf extracts with 100 ppm streptomycin stimulated germination significantly compared to Leaf extract alone but reduced root length of control by 43%. Root length was more sensitive to the autotoxin(s) than was germination or hypocotyl length. These results suggest that agar medium mixed with extract and sterilization by autoclaving could be improved the consistency and precision of bioassay, and that root length was the best parameter of autotoxic effect of alfalfa leaf extract.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.6
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• pp.821-832
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• 1997

The aim of this study was to separate or purify some bioactive compounds from flowers of alfalfa(Medicago sativa L.) and to test of the isolated compounds on alfalfa for their autotoxicity and on Echinochloa crus-galli for their allelopathy for seed germination and seedling weight. Using thin layer chromatography(TLC) of $CHCl_3$ extracts, the most inhibitory band to alfalfa seed germination was determined. Germination inhibition of this extract suggested a complex chemical interaction. Separation and purification of compounds with CHCl$_3$ extract of fresh alfalfa flowers were conducted by a silica gel TLC, and microcrystalline cellulose TLC(MCTLC), followed by droplet countercurrent chromatography(DCCC) bioassay. Preliminary identification was done by high perfomance liquid chromatography(HPLC) on the most inhibitory fractions in DCCC. Ferulic acid, caffeic acid, vanillic acid, rutin, narringin were identified in fraction 5 and ferulic acid, caffeic acid, vanillic acid, rutin, coumarin in fraction 6. The phytotoxicity of their individual compound was tested on alfalfa and Echinochloa crus-galli seed germination and seedling weight. Coumarin and ferulic acid showed the most inhibitory effect on alfalfa seed germination and Echinochloa crus-galli seedling fresh and dry weight. These compounds may be, at least in part, involved in autotoxicity and allelopathy.

• Natural Product Sciences
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• v.29 no.1
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• pp.42-49
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• 2023

This study investigated the effect of ethanol extracts of horsetail, alfalfa, ortie, chêne and aleppo oak on blood coagulation in vitro. Extraction was performed by the maceration method. Extracts were mixed with platelet and plasma, then prothrombin time (PT), activated partial thromboplastin time (APTT) and platelet aggregation tests were conducted. Alfalfa extract had a dose-dependent effect on the PT. Ortie, and horsetail, reduced the PT significantly compared to control group. Alfalfa, horsetail, and ortie reduced the APTT, but their effect was insignificant compared to the control group. The pooled extract showed the highest effect compared to the single extracts in a dose-dependent manner. Horsetail and alfalfa induced platelet aggregation in response to arachidonic acid but not in response to collagen. In the case of ortie, no aggregation occurred regarding the arachidonic acid, and incomplete was observed in response to collagen. Interestingly, blood clotting occurred immediately after adding the chêne, aleppo oak and the pooled extract, and therefore platelet poor plasma (PPP) and platelet rich plasma (PRP) became jelly. Generally, chêne and aleppo oak, as well as pooled extract, were more effective in inducing both primary and secondary coagulation pathways via shortening the PT and APTT, and induction of platelet aggregation.