• Title/Summary/Keyword: Phenol Red

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Antioxidant and Antiobesity Activity of Natural Color Resources (천연색소 소재의 항산화 및 항비만 활성)

  • Hwang, Cho-Rong;Tak, Hyun-Min;Kang, Min-Jung;Suh, Hwa-Jin;Kwon, Oh-Oun;Shin, Jung-Hye
    • Journal of Life Science
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    • v.24 no.6
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    • pp.633-641
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    • 2014
  • This study investigated the antioxidant and antiobesity activity of extract powders from the following natural color resources: Polygonum indigo, Black locust, Cochineal, Catechu, Grape, Tesu flower, Henna, Chrysanthemum, Sandalwood Red, Himalayan Rhubarb, and Madder. Total phenol content was the highest in Catechu extract, at 348.25 mg/g. DPPH, ABTS radical scavenging activity and ferric reducing antioxidant power (FRAP) were also higher in Catechu extract. Bleaching inhibition activity in the ${\beta}$-carotene linoleic acid system was the highest in Black locust extract, as was ${\alpha}$-Glucosidase inhibition activity. ${\alpha}$-Amylase inhibition activity was the highest in Catechu extract. Trypsin inhibition activity of Black locust extract was greater than 60%, and ${\alpha}$- chymotrypsin inhibition activity of Catechu extract was greater than 40%. Lipase inhibition activity was the highest Black locust extract, at 52.73%. Viability of 3T3-L1 cells was not affected by treatment with extracts at concentrations of $1.25{\sim}25{\mu}g/ml$. Lipid accumulation in the 3T3-L1 cells was the lowest following treatment with Catechu extract, at 55.8%, and this extract also inhibited adipocyte differentiation. These results suggest that the Catechu and Black locust extracts have high antioxidant and antiobesity activities and can be useful ingredients in functional foods.

Physicochemical and Sensory Characteristics of Sesame Oils Manufactured in Korea, Japan and China (한국(韓國), 일본(日本), 중국(中國) 삼개국(三個國) 참기름의 이화학적(理化學的) 특성(特性) 및 궁능적(宮能的) 특성(特性))

  • Kim, Hyeon-Wee;Lee, Min-Jung;Kim, Ki-Hong
    • Proceedings of the EASDL Conference
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    • 2004.10a
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    • pp.107-129
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    • 2004
  • Sesame oil has been popular for hundreds of years in Korea because of its pleasant flavor and health benefits and has been studied for its antioxidant properties and flavor preferences attributed to its manufacturing methods. The objective of this study was to investigate the qualitative properties of six commercial sesame oils (3 Korean, 2 Japanese, 1 Chinese), The fatty acids in the oil are composed of two main acids oleic acid and linoleic acid with a P/S ratio of 4.99${\sim}$5.73. Of the tocopherol isomers, ${\gamma}$-toc ranged from 23.14 to 34.85mg/100g. Lignan such as sesamin(322.91${\sim}$689.39ppm) and sesamolin (62.19${\sim}$289.82 ppm) is found predominantly in sesame oil. Sesamol (8.52${\sim}$51.21 ppm) was significantly different depending on manufacturer, observed as greatest in the Korean and least in the Japanese products. The induction period was longest in order of the Korean, Chinese, and then Japanese product. The red and yellow values in Lovibond color were highest in the Korean and lowest in the Japanese product. The major volatile compounds (in order of content) were pyrazines, phenols, aldehydes, and then furans and contained a small amount of pyrroles, thiazoles and indoles. The levels of total volatiles were greatest in the Korean and least in the Japanese product. The most abundant volatiles in the Korean product were pyrazines, whereas phenols were higher in the Chinese product compared to the others. From these results, the relationships among pyrazines, sesamol, yellowness and induction period showed positive, respectively. In sensory evaluation, Korean panelists preferred, in order, the Korean, Japanese, and then the Chinese product in strength of and preference for the sesame flavor, also ranking it best in overall acceptance. Japanese panelists found similarities in the Korean and Japanese products and gave an equal level of preference for the sesame flavor and overall acceptance. On the other hand, Chinese panelists preferred the Japanese product in strength and sesame flavor rating it best on overall acceptance.

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Anti-oxidant and anti-adipocyte differentiation of Aster glehni and Aster yomena (섬쑥부쟁이와 쑥부쟁이의 항산화 및 지방세포 분화 억제 효과)

  • Lee, Ji Yeon;Park, Jeong-Yong;Kim, Hyung Don;Lee, Seung Eun;Lee, Jeong Hoon;Lee, Yunji;Seo, Kyung Hye
    • Journal of Nutrition and Health
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    • v.52 no.3
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    • pp.250-257
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    • 2019
  • Purpose: Aster glehnii (AG) and Aster yomena (AY) are medicinal plants that belong to the family Compositea and grow widely in Korea. Plants in the genus Aster have been used to treat snakebite wounds or bruises in oriental medicine. This study compared the effects of anti-oxidants and anti-adipocyte differentiation according to the species (the aerial parts of AG and AY). Methods: AG and AY were extracted using 70% ethanol (-E) and water (-W) at room temperature. The anti-oxidant activities were measured by total phenol contents (TPC), total flavonoid contents (TFC), DPPH and $ABTS^+$ assay. In addition, correlation analysis was performed for the anti-oxidant compounds and effect. The level of anti-adipocyte differentiation was assessed using an oil red O assay on pre-adipocytes. Results: AG-W showed higher TPC ($6.92{\mu}g/mL$) and AG-E presented higher TFC ($8.22{\mu}g/mL$) than the other extracts. Furthermore, AG-E exhibited higher radical scavenging activity in the DPPH and $ABTS^+$ assay ($IC_{50}$: 104.88 and $30.06{\mu}g/mL$). In the cytotoxicity assay, AG and AY extracts at concentrations less than $100{\mu}g/mL$ were non toxic. AG-W reduced the lipid accumulation of 3T3-L1 cells significantly after differentiation (70.49%) compared to the other extracts. Conclusion: These results show that the water extract of AG has anti-oxidant effects and reduces the differentiation of 3T3-L1 cells. Therefore, AG has utility as a functional food material for its anti-oxidant activities and ability to prevent lipid accumulation.

The Effect of Nitric Oxide Donor or Nitric Oxide Synthase Inhibitor on Oxidant Injury to Cultured Rat Lung Microvascular Endothelial Cells (산화질소 공여물과 산화질소 합성효소 길항제가 백서 폐미세혈관 내피세포 산화제 손상에 미치는 영향)

  • Chang, Joon;Michael, John R.;Kim, Se-Kyu;Kim, Sung-Kyu;Lee, Won-Young;Kang, Kyung-Ho;Yoo, Se-Hwa;Chae, Yang-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1265-1276
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    • 1998
  • Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.

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Priming Effect of Endotoxin in Human Alveolar Macrophage (사람 폐포대식세포에서 내독소의 Priming 효과)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.46-53
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    • 1996
  • Background: Endotoxin or lipopolysaccharide(LPS) can prime phagocytic cells such as polymorphonuclear leukocytes, monocytes or animal peritoneal macrophages to generate increased amounts of secretory products such as oxygen free radicals and tumor necrosis factor, which play an important role in developing adult respiratory distress syndrome in gram negative sepsis. Human alveolar macrophages(HAM) are continuously exposed to various stimuli inhaled into the alveoli, and the response to LPS might be different in HAM. Therefore, we investigated the effect of LPS pre-exposure on HAM adhered to plastic surface and A549 cell(type II human alveolar epithelial cell line) monolayer. Methods: HAM were isolated from bronchoalveolar lavage fluid from normal lung of the patients with localized lung cancer and esophageal cancer. LPS was exposed to HAM for 2hrs before or after adherence to plastic surface of 24-well Linbro plate and A549 cell monolayer. And then HAM was stimulated with PMA(phorbol myristate acetate) or fMLP(N-formyl-methionylleucyl-phenylalanine). The amount of hydrogen peroxide($H_2O_2$) production in the supernatant was measured on the principle of peroxidase-dependent oxidation of phenol red by hydrogen peroxide. Results: LPS pre-exposure could not enhance $H_2O_2$ production in neither HAM adhered to plastic surface nor one to A549 cell monolayer. But LPS even in the absence of PMA or fMLP stimulation directly increased $H_2O_2$ release in HAM if added after the adherence to A549 cell monolayer. Conclusion: Endotoxin does not prime HAM, but may directly activate HAM adhered to alveolar epithelial cells. Further investagation will be necessary.

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Lymphocyte Subpopulations and Proliferation of T cells, Phagocytic Activity of Leukocytes on Alcoholics (알코올중독자의 백혈구탐식능, 림프구아형 및 증식능)

  • 김용호;서병배;이정녀;김영훈
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.167-174
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    • 1996
  • Alcoholics increased susceptibility to microbial infection that is associated with decreased immunity. but there has been little experimental evidence to support alcoholics-induced increase of microbial infection directly in non-specific immunity. Therefore, we were used the method of phagocytic-plaque including all the stimulating factors for the phagocytosis, subtypes of lymphocytes and T-lymphocyte proliferation. The experimental groups were divided into 3 groups: (1) alcoholics who were hospitalized less than 1 week (newly hospitalized alcoholics), (2) alcoholics who were hospitalized more than 2 weeks (old hospitalized alcoholics), (3) healthy blood donors. We have studied 98 alcoholics and 35 healthy blood donors and control groups. A physician has checked the biological markers and diagnosed the body-condition alcoholics. The immunity and non-specific immunity on the alcoholics were analyzed by using the simultest kit and flow cytometry. Proliferation of the lymphocytes was analyzed by the phytohemmagglutinine mitogen. Phagocytosis and migration properties of leukocytes were identified on the layer formed by Staphylococcus aureus Cowan I strain. Biological markers of alcoholics and control groups, by such as blood glucose, ${\gamma}$-glutamyl transpeptidase and mean corpuscular volumes of red blood cells, were determined by biochemical and hematological methods. Compared with control groups, cluster of differentiation (CD)3+, CD8+ and CD19+ in alcoholic were more decreased except CD4+/CD8+ ratio. Proliferation of the T-lymphocytes, phagocytosis and migration properties of the leukocytes in alcoholics were decreased compared with those of control groups. According to the results observed in our experiment, they can be summerized as follows: 1, Cellular, humoral and non-specific immunities, are markedly decreased in alcoholics than those in control groups. 2. It is inferred that Phagocytic plaque formation is a very useful method to evaluate phagocytosis and migration properties of the alcoholic leukocytes 3. It is thought that the subtypes of lymphocytes, especially CD4+/CD8+ ratio, are essential methods to analyzed the alcoholic immunity. 4. Specific and non-specific immunity on the old hospitalized alcoholics was slightly increased, which depends upon the alcoholic medication.

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Studies on the Processing of Low Salt Fermented Sea Foods 10. Changes in Volatile Compounds and Fatty Acid Composition during the Fermentation of Yellow Corvenia Prepared with Low Sodium Contents (저식염 수산발효식품의 가공에 관한 연구 10. 저식염조기젓 숙성중의 휘발성성분 및 지방산조함의 변화)

  • CHA Yong-Jun;Lee Eung-Ho;PARK Du-Cheon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.6
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    • pp.529-536
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    • 1986
  • By modified method yellow corvenia(called $Y_3$) was prepared with $4\%$ salt, $4\%$ KCl, $6\%$ sorbitol, $0.5\%$ lactic acid and $4\%$ alcohol extract of red pepper to improve the quality of fermented sea food. In this study, changes of volatile compounds and fatty acid composition obtained from modified fermented yellow corvenia($Y_3$) were experimented during fermentation, comparing with conventional fermented yellow corvenia(called $Y_1,\;20\%$ of salt contents). Total lipid of yellow corvenia was composed of $78.1\%$ of neutral lipid, $21.2\%$ of phospholipid and $0.7\%$ of glycolipid. And monoeonoic acid was held $37.4\%$ of fatty acid composition of total lipid and saturated fatty acid ($34.8\%$), polyenoic acid ($27.7\%$) were followed. Saturated fatty acid($C_{14:0},\;C_{16:0},\;C_{18:0}$) in $Y_1,\;Y_3$ increased, polyenoic acid ($C_{22:6}\;C_{22:5}\;C_{20:5}$) decreased while monoenoic acid($C_{16:1}\;C_{18:1}$) in those was little fluctuated during fermentation. Thirty-three kinds of volatile component in whole volatile compounds obtained from $Y_1,\;Y_3$ at 90 days fermentation were identified, and composed of some hydrocarbons (8 kinds), alcohols (7 kinds), acids (6 kinds), aldehydes(4 kinds), sulfides(2 kinds), ketones (2 kinds), one of phenol and 3 kinds of the other components. Among the whole volatile compounds 2-ethoxy ethanol and was held $79.35\%$ in $Y_3$, whereas nonadecane was held $75.85\%$ in $Y_1$. During fermentation 8 kinds of volatile acids, 5 kinds of amines and 9 kinds of carbonyl compounds were also detected. Those volatile acid such as acetic acid, isovaleric acid, n-caproic acid, n-butyric acid were the major portion of total volatile acids in $Y_3$ at 90 days fermentation. Meanwhile, carbonyl compounds such as ethanal, 2-butanone and butanal were the major ones, while TMA held the most part of volatile amines in $Y_3$ during fermentation. From the result of sniff test, the components which are believed to contribute to the characteristic flavor of fermented product $Y_1,\;Y_3$ are deduced to be volatile acid, carbonyl compounds and amines in order. Conclusively, there was little difference in composition of volatile components, but merely a little difference in content of those between $Y_3$ and $Y_1$.

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Suppressive Effect of Green Tea Seed Coat Ethyl Acetate Fraction on Inflammation and Its Mechanism in RAW264.7 Macrophage Cell (RAW264.7 Macrophage Cell에서 녹차씨껍질 에틸아세테이트 분획의 염증억제 효과 및 기전 연구)

  • Noh, Kyung-Hee;Jang, Ji-Hyun;Min, Kwan-Hee;Chinzorig, Radnaabazar;Lee, Mi-Ock;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.5
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    • pp.625-634
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    • 2011
  • Green tea seed coat (GTSC) was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether (PE), ethyl acetate (EtOAC) and butanol (BuOH). The EtOAC fraction showed the highest level in total phenol contents and the lowest level in nitric oxide (NO) production in LPS-stimulated RAW264.7 macrophage cell. Thus, this study was carried out to investigate the anti-inflammatory and its mechanisms of GTSC EtOAC fraction in LPS-stimulated RAW264.7 macrophage cell. GTSC EtOAC fraction contained EGC ($1146.48{\pm}11.01\;{\mu}g/g$), tannic acid ($966.99{\pm}32.24\;{\mu}g/g$), EC ($70.88{\pm}4.39\;{\mu}g/g$), gallic acid ($947.61{\pm}1.03\;{\mu}g/g$), caffeic acid ($37.69{\pm}1.46\;{\mu}g/g$), ECG ($35.46{\pm}3.19\;{\mu}g/g$), and EGCG ($15.53{\pm}0.09\;{\mu}g/g$) when analyzed by HPLC. NO production was significantly (p<0.05) suppressed in a dose-dependent manner with an $IC_{50}$ of $80.11\;{\mu}g$/mL. Also prostaglandin $E_2$ level was also inhibited in a dose-dependent manner. Moreover, iNOS protein expression was suppressed in dose-dependent manner but COX-2 gene expression was not affected. Total antioxidant capacity and glutathione (GSH) levels were enhanced more than the LPS-control. Expressions of antioxidative enzymes including catalase, GSH-reductase and Mn-SOD were elevated compared to LPS-control. Nuclear p65 level was decreased in the GTSC EtOAC fraction in a dose-dependent manner. These results indicate that GTSC EtOAC fraction inhibit oxidative stress and inflammatory responses through elevated GSH levels, antioxidative enzymes expressions and suppression of iNOS expression via NF-${\kappa}B$ down-regulation.

Antioxidant activities of commonly used Brassica spp. sprout vegetables in Korea (국내 다소비 십자화과 새싹채소 추출물의 항산화 활성)

  • Shin, Gi-Hae;Lee, Young-Jun;Kim, Jae-Hwan;Kim, Young-Hyoun;Kim, Dan-Bi;Lee, Jong Seok;Lim, Jeong-Ho;Lee, Ok-Hwan
    • Food Science and Preservation
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    • v.21 no.4
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    • pp.587-592
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    • 2014
  • Brassica spp. vegetables have been known to have biological activities such as anti-cancer and anti-inflammatory activities. In this study, we investigated the in vitro physicochemical characteristics and antioxidant activities of commonly used Brassica spp. sprout vegetables such as kohlabi (Brassica oleracea var. gongyloides), red radish (Raphanussativus L. var. sativus), broccoli (B. oleracea var. italica), cabbage (B. rapavar. glabra Regel), rape (B. napus), radish (R. sativus), and tatsoi (B. campestris var. narinosa) sprouts. Our results showed that the vegetables with the highest total phenolics contents were the radish sprout ($24.40{\pm}1.24mg\;TAE/g$) and kohlabi sprout extracts ($23.97{\pm}0.46mg\;TAE/g$). Furthermore, the vegetable with the highest total flavonoid content was the radish sprout extract ($15.30{\pm}1.35mg\;CE/g$). However, the kohlabi sprout extract showed the highest DPPH radical scavenging value ($IC_{50}=1.95mg/mL$) and ORAC value (79.03 mM TE/g). In addition,the six kinds of Brassica spp. sprout vegetable extracts, except tatsoi, significantly inhibited the reactive oxygen species (ROS) production and showed that intracellular oxidative stress is closely related tothe accumulation of differentiated adipocytes and fat during the adipogenesis of 3T3-L1 preadipocytes. These results suggest that Brassica spp. sprout vegetables, especially kohlabi and radish sprout extracts, can be used to develop natural antioxidants.

Studies on the Species Crossabilities in the Genus Pinus and Principal Characteristics of F1 Hybrids (일대잡종송(一代雜種松)의 교배친화력(交配親和力)과 특성(特性)에 관(關)한 연구(硏究))

  • Ahn, Kun Yong
    • Journal of Korean Society of Forest Science
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    • v.16 no.1
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    • pp.1-32
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    • 1972
  • By means of the interspecific hybridization in the Sub-genus Diploxylon of the Genus Pinus, $F_1$ hybrids of Pinus rigida${\times}$elliottii, Pinus rigida${\times}$radiata, P. rigida${\times}$serotina and P. densiflora${\times}$thunbergii had been produced. And on the basis of the crossabilities of these hybrids the taxonomic affinities of these pines were examined. And the needle characteristics of these hybrid and the occurence of phenolic substances in these $F_1$ hybrid were also investigated to see the potential usefulness of these characteristics for the diagnosis of the taxonomic affinity. And, the growth performances of the $F_1$ hybrids have also been compared with those of parental species. In order to contribute to the establishment of the hybrid seed orchard the introgression phenomena between P. densiflora and P. thunbergii in the eastern coastal area have also been investigated along with the investigation of the heterozygosity of plus trees of P. densiflora growing in the clone bank in Suwon. And the results were summarized as follows. 1. On the basis of crossabilities as well as on the taxonomic affinities according to the systems of Shaw, Pilger and Duffield, it has been proven that the parental species of those hybrids are of close affinities and range of the fertile hybrid seed production rate was as high as 28-58% in the best hybrid combination (Table 13). 2. Among those hybrids, the ${\times}$ Pinus, rigiserotina hybrid seemed to be most promising in the growth performance exhibiting 109-155% more volume growth compared to the seed parent with the statistic significance of 1% level (Tables 16 and 17). 3. Notwithstanding the fact that the all of the pollen parents are cold tender, all hybrids exhibit cold hardiness as much as their seed parent and it seems to suggest that the characteristics of cold hardiness were transmitted from the seed parent. 4. Though a striking difference in needle length was observed between the parental species of each hybrid, it was difficult to distinguish each hybrid from their seed parent by the needle length except ${\times}$P. rigiserotina which is characterized by long needle which is 65% more longer than the needle of the seed parent (Table 21). 5. With regard to the anatomical characteristics of needle, the hypoderm is apparently thicker in most of the $F_1$ hybrid pines and the characteristics of resin canals are dominated by medial in most $F_1$ hybrid. And, the fibrovascular bundles were apart as were in their seed parent. Therefore it was found to be possible to distinguish the hybrids pines from their parents by the needle characteristics. And, it is to be noticed that the ${\times}$P. densithunbergii was more close to the pollen parent having RDI value of 0.73 (Fig.l, Table 22). 6. It has been demonstrated that ${\times}$P. rigielliottii, ${\times}$P. rigiradiata and ${\times}$P. rigitaeda have a phenolic substance (No.7) of light yellow at Rf-0.46, same as their seed parent, but no trace of phenolic substance was observed in their pollen parent. This fact will serve as an important criteria for early identification of hybridity in progeny testing. However, the fact that both of ${\times}$P. rigiserotina and ${\times}$P. densithunbergii exhibit the same reactions of phenolic substances as well their parental species seems to indicate the close affinities between the parental species of the respective hybrid (Fig.2, Table 23). 7. The separation and the reaction of phenolic substance developed on TLC were found to be same in the same species showing no variations between the individuals, and no variations due to tree part of sampling, tree age or pollen sources. And the reaction was also observed regardless of the not varied by the kind of developing solvent whether it is Aceton-Chloroform (3:7 v/v) or Benzene-Methanol-Acetic acid (90:16:8 v/v). 8. The introgression phenomena of natural Pinus densifiora stand in both east and west coastal area indicates that the major part of the red pines investigated are all heterozygous and the heterozygosity of pines are higher in the west coast than in the east coast(Tables 24 and 25). 9. Based on the RDI, among the plus trees of Pinus densiflora selected in Korea and Japan as well, no pure P. densiflora has been found. Since all of the sample trees of Pinus densiflora were found to be as heterozygous bearing part of the characteristics of P. thunbergii, those red pines were considered to be natural heterotic hybrid pines(Figs. 3 and 4. Tables 26 and 27).

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