• Analytical Science and Technology
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• v.7 no.3
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• pp.403-411
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• 1994

To isolate, purity and identify of bioactive compounds involved in alfalfa allelopathy and/or autotoxicity, experiment was conducted. Isolation and separation procedures used from an 80% methanol extract of fresh alfalfa leaves(1kg), silica gel thin layer chromatography(TLC), followed by Droplet Counter Current Chromatography(DCCC). Preliminary identification was examined by high preformance lipid chromatography(HPLC). Four phenolic compound, salicylic acid, scopoletin, rutin, and quercetin, were identified and identified all compounds were phytotoxic to alfalfa seed germination and seedling growth. Among these compounds, quercetin treatment($10^{-3}M$) was most inhibitory to alfalfa seed germination and seedling growth. These compounds may be, at least in part, involved autotoxicity and allelopathy.

• 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.

• Proceedings of the Korean Society of Crop Science Conference
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• 1995.05a
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• pp.64-65
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• 1995

• Plant Resources
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• v.3 no.1
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• pp.71-78
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• 2000

Autotoxicity restricts reseeding of new alfalfa(Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments, often requiring up to a year or more. One solution for reducing autotoxicity would be to select germplasms or cultivars with tolerance to the autotoxic chemical(5) and use genetically breeding program. Bioassay of seed germination and early seedling growth was conducted to evaluate autotoxic responses of 3 varieties of alfalfa to the water-soluble extracts(at 4 and 8g/L) from alfalfa ‘Cody’leaf by using agar and filter paper medium in a petri-dish assay. Root length at 5 days after seeding was more sensitive to the extract than was hypocotyl length or seed germination, and was a better parameter of autotoxic effects of alfalfa leaf extracts. Use of an agar medium gave better sensitivity of root length than did use of filter paper. Evaluating tolerance with percent of control was more important indicator than was mean of root length because of significant variation among varieties in root length of control treatment. Bioassay ranked varieties in the following order of tolerance on the basis of relative root length; “Cody” >“ Pioneer 5373” >“ Alfagraze”. Seedling growth from old “Cody” seed was more sensitive to the autotoxic chemical(5) than was that from newly produced seed.

• Proceedings of the Korean Society of Crop Science Conference
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• 1994.06a
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• pp.88-89
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• 1994

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.2
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• pp.134-138
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• 2000

Most parts of alfalfa plant have been reported to contain autotoxic substances that inhibit seed germination and early seedling growth, however, the chemical(s) is not still studied much. Effect of seed leachates of 'Vernal' alfalfa (Medicago sativa L.) was evaluated for inhibition of alfalfa germination and root growth through bioassay. Alfalfa seeds were extracted in 1 L deionized water for 1 h after soaking and the leachates caused to reduce root length of alfalfa significantly as the soaking time increased. Crude seeds at 4 g L$L^{-1}$ exudated autotoxins that reduce significantly root length by 34 % compared to the control, when the seeds soaked in deionized water for 24 h. However, the extracts did not affect final germination as well as speed of germination. Extracts from ground seeds significantly reduced speed of germination (GT 50) and root length. The results indicate that release of autotoxic substances from seeds during seed imbibition was increased with increase of soaking time and seed amount, and that autotoxicity was more occurred in ground seeds than in crude seeds.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.2
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• pp.228-235
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• 1997

This experiments were conducted to determine the effect of allelopathy and autotoxicity in alfalfa. Among several alfalfa cultivars, the substances exudated from seven cultivars including Medicago sativa, c.v. Luna, Sparta, Magnum, Husky, Milkmaker, Challenger and Anchor inhibited the germination and seedling growth of radish, ranging from 60 to 80％. When allelopathic substances were exudated from the soil of the alfalfa-cultivated field, the increased levels of substance retarded the germination rate and seedling growth of radish and alfalfa. The inhibition rate was about 80％, compared with that of control. Thus, this study indicates that the allelopathy and autotoxicity substances of alfalfa influenced the germination and seedling growth in alfalfa itself and radish. When allelopathic substances were analysed by using gas chromatograph, several kinds of phenol compounds were detected as follows; Salicylic acid, hydroxybenzoic acid, vanillic acid, syringic acid, coumaric acid, and ferulic acid.

• 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.

• Journal of Forest and Environmental Science
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• v.26 no.2
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• pp.83-94
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• 2010

Chinese fir (Latin name: Cunninghaimia lanceolata) is one of the major commercial coniferous trees. Most of Chinese fir forests are managed in successive rotation sites, which lead productivity to decline. Autotoxicity is the important reason for soil degradation of Chinese fir plantation, especially, phenolic acids are considered as the major allelopathic toxins which induce autotoxicity in Chinese fir rotation stands. We performed here proteomic approach to investigate the response of proteins in Chinese fir leaves to salicylic acid. The tube plantlets of Chinese fir clone were treated with 120 mg/L salicylic acid for 1, 3 and 5th day. 2-DE, coupled with MALDI-TOF-TOF/MS, was used to separate and identify the responsive proteins. We found 12, 7, and 12 candidate protein spots that were up- or down-regulated by at least 2.5 fold after 1, 3, and 5th day of the stress, respectively. Of these protein spots, 16 spots were identified successfully. According to the putative physiological functions, these proteins were categorized into five classes (1) the proteins involved in protein stability and folding, including 26S proteome, Grp78, Hsp70, Hsp90 and PPIase; (2) the protein involved in photosynthesis and respiration, including OEC 33 kDa subunit, GAPDH; (3) the protein related to cell endurance to acid, F-ATPase; (4) the protein related to cytoskeleton, tubulin; (5) the protein related to protein translation: prolyl-tRNA synthetase. These results give new insights into autotoxic substance stress response in Chinese fir leaves and provide preliminary footprints for further studies on the molecular signal mechanisms induced by the stress.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.299-302
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• 2000

Actinoplanes teichomyceticus ATCC 31121 was mutated with UV to obtain a superior mutant strain with increasing level of teicoplanin production. In this investigation lethal curve was obtained and the optimal condition to induce mutagenesis was determined and to isolate the desirable mutant strain. It was also confirmed that teicoplanin activities by agar diffusion method to be compared to the mother strain. One mutant strain, T991014-1 that had the highest teicoplanin productivity was finally selected for further investigation including fermentation pattern. The mutant was characterized by the various tests such as amylase activity, protease activity, antibiotic resistance, autotoxicity, and productivity.