• Title/Summary/Keyword: Stage I

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THE ASPECT OF PROLIFERATION AND BONE NODULE FORMATION IN OSTEOBLAST-LIKE CELLS DERIVED FROM FETAL RAT CALVARIA IN VITRO (백서 태자 두 개관에서 유래된 조골세포의 증식 및 골결절 형성양상)

  • Kim, Shi-Hyeong;Nam, Soon-Hyeun;Shin, Hong-In
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.1-17
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    • 1997
  • The purpose of this study was to investigate the aspects of proliferation and bone nodule formation of osteogenic precursor cells. To determine the effects of ascorbic acid and dexamethasone upon capacity of osteoblast proliferation and bone nodule formation, cells were maintained in the presence of one or some of these additives for up to 30 days. Group I culture was maintained in standard medium(DMEM plus 10% plus antibiotics), group II was maintained in supplemented medium containing dexamethasone, group III was maintained in supplemented medium containing ascorbic acid and sodium-${\beta}$-glycerophosphate, and group IV was maintained in supplemented containing ascorbic acid, sodium-${\beta}$-glycerophosphate and dexamethasone. Morphology of bone nodules was observed with light microscope and electron microscope. The results were as follows: ${\bullet}$ Proliferation capacity of osteoblasts was not affected by single use of dexamethasone, but it was chiefly affected by ascorbic acid. ${\bullet}$ Cellular morphology was fibroblastic appearance initially, but, it was gradually changed to polygonal shape accompanied by confluency stage. ${\bullet}$ Pluripotent mesenchymal cells existed during primary culture, they were differentiated to adipocyte, chondrocyte, osteocyte according to culture condition. ${\bullet}$ Dexamethasone increased bone nodule formation under the condition that the culture was maintained with supplemented medium ascorbic acid and sodium-${\beta}$-glycerophosphate. ${\bullet}$ when the cultures were stained with alizarin red, the group supplemented with dexamethasone, ascorbic acid and sodium-${\beta}$-glycerophosphate showed the marked increase of bone nodule formation, but the group supplemented with ascorbic acid and sodium-${\beta}$-glycerophosphate revealed only small amounts of bone nodules. And the groups cultured without ascorbic acid showed no observed any of bone-like mass independent of dexamethasone addition.

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The Effect of Surface Sealing on the Microleakage of Class V Composite Resin Restorations (제V급 복합레진 수복물의 표면전색이 미세변연누출에 미치는 효과)

  • Youn, Yeon-Hee;Hyun, Hong-Keun;Kim, Young-Jae;Kim, Jung-Wook;Jang, Ki-Taeg;Lee, Sang-Hoon;Kim, Chong-Chul;Hahn, Se-Hyun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.3
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    • pp.359-369
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    • 2007
  • This in vitro study was performed to assess the effect of surface sealing on the microleakage of class V composite resin restorations that underwent several aging treatments. Class V cavities were prepared on the buccal surface of 100 sound extracted premolars and restored with a hybrid light-cured composite resin according to the manufacturer's instructions. They were randomly divided into two groups consisting of 50 samples: group I, without surface sealing, and group II, in which margins were etched and surface sealant was applied. After thermocycling, each group was divided into five subgroups, respectively, to represent the five aging treatments: group A = no further treatment (only thermocycling), B = toothbrushing, C = load cycling, D = toothbrushing followed by load cycling, and E = aging treatment in deionized water for six months. Microleakage was assessed by examining the penetration of 2% methylene blue dye. The following results were obtained: 1. At occlusal and cervical margins in groups without surface sealing, there was no significant difference in microleakage after the several aging treatments (p>0.05). 2. The occlusal margins of groups with surface sealing showed no significant differences after the several aging treatments (p>0.05). 3. In the cervical margins of groups with surface sealing, microleakage significantly increased after load cycling or aging in deionized water for six months (p<0.05). 4. The no-further-treatment group and the toothbrushing group with surface sealing showed less microleakage than the corresponding groups without surface sealing (p<0.05). 5. The surface-sealed groups with load cycling or aging in deionized water showed no significant difference in microleakage to the corresponding groups without surface sealing (p>0.05). In conclusion, the results of this study suggest that the surface sealant infiltrating through the gap of the cervical margin exerted a positive effect on microleakage at the initial stage, but the effect was not sufficient to overcome the stress generated by the cuspal flexure during occlusal loading and water absorption.

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Effect of Nursery stage and Plug Cell Size on Seedling Growth of Waxy Corn (찰옥수수 육묘일수와 플러그셀 크기가 묘의 생육에 미치는 영향)

  • Kim, Sung-Kook;Jung, Tae-Wook;Lee, Yu-Yong;Song, Duk-Yong;Yu, Hong-Seob;Lee, Choon-Woo;Kim, Yee-Gi;Kwak, Chang-Gil;Jong, Seung-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.407-415
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    • 2009
  • To investigate the effects of seeding date, nursery days and plug cell size on seedling growth and quality of waxy corn. This study was carried out in the field of the National Institute of Crop Science in 2007. Varieties used were two waxy corn hybrids cvs. Chalok# 1 and Chalok# 4. The plant height, number of leaves and dry weight of waxy corn seedlings were significantly affected by seeding dates and plug cell size, while no differences between varieties. The growth of seedlings was faster in March 14 seeding than in May 8 and July 4 seedings. Based on plant height, March 14 seeding, May 8 seeding and July 3 seeding reached suitable size for transplanting after 30 days (21.6cm), 15 days (25.5~26.9cm) and 10 days (21.6~24.0cm) in the nursery, respectively. Seedlings of March 14 seedling and May 8 and July 3 seeding reached suitable number of leaves i,e. 2~3 leaves, 15~20 days after seeding on March 14 and 10 days after seeding on May 8 and July 3. Dry weights of 30 days seedling for March 14 seeding, 20 days seedling for May 8 seeding and 15 days seedling for July 3 seeding were 0.43g, 0.57~0.67g and 0.53g, respectively, and the dry weight of seedlings increased by 2~3 folds at the later nursery days. The suitable nursery days of waxy corn were 20 days on March 14 seeding and 10~15 days on May 8 or July 3 seeding.

Effect of Genistein on the Onset of Puberty in Female Rats (암컷 흰쥐의 사춘기 개시에 미치는 Genistein의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.1
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    • pp.55-61
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    • 2006
  • There is growing concern that dietary soy intake is associated with protection of breast cancer. However, questions persist on the potential adverse effects of the main soy constituent genistein(GS) on female reproductive physiology. In this study, we examined whether prepubertal exposure to GS affected on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. GS(100mg/kg/day) was administrated daily from postnatal day 25(PND 25) to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the day after VO occurred. Gross anatomy and tissue weight were compared to test the GS's effect on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in tissues. Specific radioimmunoassay(RIA) were carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptors(PR), total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a results, advanced VO was shown in the GS group(PND $31.2{\pm}0.6$) compared to the vehicle group (PND $35.3{\pm}0.7$). GS treatment significantly increased wet weight of ovaries and uteri compared to the vehicle group. Increased serum LH levels were also shown in the GS group. Graafian follicles and corpora lutea(CL) were observed only in the ovaries from GS treated animals. Similarly, hypertrophy of luminal and glandular uterine epithelium were found only in the GS group. Collectively, these effects were probably due to the estrogenic effects of GS. In the semi-quantitative RT-PCR studies, the transcriptional activities of PR in both ovary and uterus from GS-treated group were significantly higher than those from the vehicle group. The present studies demonstrated that acute exposure to GS, at levels comparable to the ranges of human exposure, during the critical period of prepubertal stage activates the reproductive system resulting precocious puberty in immature female rats.

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Development of an Efficient Screening Method for Resistance of Chili Pepper Plants to Meloidogyne incognita (뿌리혹선충 Meloidogyne incognita에 대한 저항성 고추를 선발하기 위한 효율적인 검정법 확립)

  • Hwang, Sung Min;Jang, Kyoung Soo;Choi, Yong Ho;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.282-293
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    • 2016
  • Root-knot nematodes (Meloidogyne spp.) are major plant pathogens that cause reductions in yield and quality of several solanaceous crops, including pepper (Capsicum spp.). These losses can be averted through planting of resistant cultivars. Plants are defined as resistant when they suppress nematode reproduction. In this study, the resistance degrees of 102 commercial cultivars of chili pepper (Capsicum annuum) to a root-knot nematode, Meloidogyne incognita, were evaluated by comparing the number of egg masses on their roots to those of 'PR huimangchan', a highly susceptible cultivar that exhibited the most egg masses of the chili pepper cultivars evaluated. Among these cultivars, forty-four (43.1%) showed resistance to M. incognita and eighteen (17.6%) were moderately resistant. The other cultivars (39.3%) were determined to be susceptible. For further study, six chili pepper cultivars (i.e., Gangryeokjosenggeon, Shinsegae, Muhanjilju, PR Bulrocho, PR Huimangchan, and Jjang) with different levels of resistance to the nematode were selected. Changes in resistance of the six cultivars under several conditions, such as inoculum concentration, plant growth stage, and cultivation period after transplanting were investigated. We found that an efficient screening method for resistance of chili pepper to M. incognita is to transplant the chili pepper seedlings 7 days before inoculation, to inoculate 28-day-old plants with M. incognita by loading 5,000 eggs per plant into the pot of soil, to cultivate the plants in a greenhouse ($25{\pm}5^{\circ}C$) for 45-60 days, to measure the number of egg masses on roots of the seedlings, and then to determine the resistance response of the plants by comparing the number of egg masses on the roots with a reference-susceptible cultivar 'PR huimangchan'.

Systems for Production of Calves from Hanwoo(Korean Cattle) IVM/IVF/IVC Blastocyst II. Simple, Efficient and Successful Vitrification of Hanwoo Blastocyst (체외생산된 한우 배반포기배로부터 송아지 생산을 위한 체계 II. 한우 배반포기배를 간편하면서 효율적이고 성공적으로 유리화 동결하는 방법)

  • Kim, E.Y.;Kim, D.I.;Park, N.H.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.281-291
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    • 1999
  • The objective of this study was to optimize the freezing/thawing method of in vitro produced Hanwoo blastocysts. Day 7 blastocysts after IVF were vitrified using EFS40 (40% ethylene glycol, 18% ficoll, 0.3 M sucrose and 10% FBS added m-DPBS) as a freezing solution and electron microscope (EM) grid (V-G) or straw (V-S) as an embryo container. In both method, freezing/thawing were treated by 2-step, treatment time was required in V-G method and V-S method, for 2 min / 3 min and 3.5 min / 10 min, respectively. Embryo survival was assessed as re-expanded and hatched rates at 24 h and 48 h after warming, respectively. The results obtained in these experiments were summarized as follows: when the effect of exposure in vitrification solution and chilling injury from freezing procedure on in vitro produced expanded blastocysts were examined, at 24 h after warming, embryo survival in exposure group (100.0%) was not different compared to that in control group (100.0%), although those results were significantly different with two vitrified groups (V-G: 87.8, V-S: 77.8%) (P<0.001). However, at 48 h after warming, hatched rates of V-G group (67.8%) were significantly higher than those of V-S group (53.3%) (P<0.05). In addition, this hatched rate in V-G group was not different with that in exposure group (73.3%). When the effects of embryo developmental stage (early, expanded and early hatching blastocysts) and embryo container (EM grid and straw) to the in vitro survival of vitrified-warmed day 7 Hanwoo blastocysts were simultaneously examined, fast developed embryos were indicated the better resistance to freezing than delayed developed one, irrespective of embryo containers (early; 57.1 & 24.4%, expanded; 84.7 & 60.6%, early hatching; 91.7 & 80.0%) (P<0.001). Especially, in expanded and early hatching blastocysts, embryo survival of V-G group (67.8, 95.0%) was significantly higher than those of V-S group (53.0, 65.0%) at 48 h post warming, respectively (P<0.05, P<0.001). Therefore, this study indicates that Hanwoo blastocysts can be cryopreserved more simple, efficient and successful by vitrification method using EM grid.

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Effect of Trehalose and Sugar-addition on the Survival Rates of Bovine IVM/IVF Embryos after Vitrification (Trehalose 와 당첨가가 우 체외 수정란의 초자화 동결에 미치는 영향)

  • 양부근;김준국;정희태;박춘근;김종복;김정익
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.307-317
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    • 1998
  • This study was to investigate the vitrification method for cryopreservation technique of bovine in vitro fertilization(IVF) embryos. The morphological appearance and viability following vitrification of IVF bovine blastocysts and expanded blastocysts were examined. Embryos obtained 6, 7, 8 or 9 days after IVF were vitrified in both 40% ethylene glycoI(EG) plus 0.3M trehalose and 20% polyvinylpyrrolidone(PVP) in DPBS(ETP, Exp. 1) and ETP solution added 0.375M dextrose (ETPD, Exp. 2). The viability of Days 6, 7, 8 and 9 vitrified /thawed embryos at 24∼48 h culture after thawing was 11.9%, 19.8%, 23.4% and 15.3% in ETP(Exp. 1), and 34.6%, 54.5%, 37.9% and 13.0% in ETPD(Exp. 2), respectively. The viability of vitrified embryos produced from the culture days after IVF did not differ in Exp. 1, but significantly differ in Exp. 2(P<0.05). The viability of blastocysts and expanded blastocysts significantly differed(P<0.05) in 15.2% and 23.3%(Exp. 1), and 25.0% and 45.8%(Exp. 2). The result of Exp. 1 was similar to that of Exp. 2 on the viability of embryo according to developmental stages, but ETP solution plus sugar(dextrose) was increased the viability of vitrified embryos. In Experiment 3, The viability of vitrified embryos was not different between 12% and 20% PVP concentrations in ETP solution according to culture days or developmental stages. To investigate the effect of addition of sugar, two type of carbohydrates and a mixture of cryoprotectants for vitrification on the survival of bovine IVF embryos, bovine Days 7 to 9 blastocysts and expanded blastocysts were cryopreserved in either 20% glycerol plus 20% EG, 0.375M sucrose and 0.375M dextrose (GESD, Exp. 4) or 20% glycerol plus 20% EG, 0.3M trehalose and 20% PVP(GETP, Exp. 5) in DPBS. Survival rates of Day 7, 8 and 9 embryos at 24∼48h culture after thawing were 71.4%, 94.6% and 40.5% in GESD, and 59.5%, 81.5% and 62.5% in GETP, respectively. Hatching rates of Day 7, 8 and 9 embryos after thawing were 28.6%, 35.1% and 16.2% in GESD, and 27.0%, 33.3% and 18.8% in GETP, respectively. These results indicates that a mixture of cryoprotectants(glycerol and EG) and addition of sugar can improve the survival rates of the bovine IVF embryos(Day 7 or 8) vitrified, and the expanded blastocyst embryos are more suitable for vitrification than early blastocysts stage.

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Effects of Wilting and Additives on the Fermentation Chrateristics , Quality and Aerobic Stability of Rye Silage (예건 및 첨가제가 호밀사일리지의 발효특성과 사료가치 및 호기적 안정성에 미치는 영향)

  • 이광녕;김동암
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.17 no.2
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    • pp.187-198
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    • 1997
  • This experiment was conducted to evaluate effects of wilting and additives on fermetation characteristics, quality and aerobic stability of silage of rye(Seca1e cereale L.) harvested at the early heading stage. Harvested rye was wilted or mixed with beet pulp and then treated with formic acid or lactic acid bacteria(LAB) inoculant. Treatments were consisted of direct cut(DC), wilted(WT), beet pulp added(BP), direct cut and formate applied(DF), wilted and formate applied(WF), beet pulp and formate applied(BF), direct cut and LAB inoculated(DL), beet pulp added and LAB inoculated(BL), and wilted and LAB inoculated(WL) silages. Afler 70 days of ensiling period, the silages were opened and exposed to air for 7 days, and the silages of opening day(0 day) were compared with the silages exposed to air for 7 days(7 day). 1. LAB inoculated rye silages(DL, WL, BL) lowered pH values effectively eom the third day of ensiling and showed most stable pattern of pH changes during the initial fermentation process. Direct cut and formate applied(DF), beet pulp and formate applied(BF) and beet pulp added(BP) silages were also effective in lowering pH from the seventh day thai direct cut treatment(DC). 2. pH was low below 3.7 in all treatments of 0 day. After 7-day period of aerobic exposure, DL and WL showed considerable increases in pH from 3.5 and 3.4 to 8.3 and 6.4, respectively. 3. Direct cut rye silages(DF, DC, DL) and beet pulp and formate treatment(BF) produced effluent of 121.2, 85.9, 80.3 and 34.2 mlkg, respectively and these were greater than others(P<0.01). Beet pulp retained 1.61 I/kg of effluent at the application rate of 50 kg/t in comparing BP with DC, and formate application increased the amount of effluent in comparing DF and BF with DC and BP, respectively(P<0.01). 4. Beet pulp and LAB treatment(B1) was highest in content of lactate as 3.1% and formate treated silages(DF, WF, BF) showed lower content of lactate and total acid than others. LAB inoculated(D1, WL, BL) and wilted (WT) silages were graded to be good quality as a second group by the Flieg's score. Formate applied silages (DF, WF, BF) were graded as a foum group lower than DC by one. 5. Residual content of WSC of rye silage was higher in BP, BF, DL and BL than direct cut treatment(M3) (P< 0.01). Formate application had a tendency to increase the content of residual content of WSC. 6. For the ratio of NH3-N to Total N, wilted and LAB inoculated(W1) and direct cut and LAB inoculated(D1) silages were lowest on 0 day as 8.9 and 9.3% respectively. But after 7-day period of aerobic exposure, WL and DL showed largest increase of the ratio of $NH_3-N$ to Total N from 0 day(P<0.01). 7. On 0 day wilted(WT), beet pulp and formate treatment(BF), and beet pulp and LAB treatment(B1) were lower than direct cut treatment(DC) for NDF and ADF concentrations consistently. Formate applied silages (DF, WF, BF) made little change in ADF and NDF concentrations during 7 day period of aerobic exposure. DL and WL showed a large increase in ADF and NDF concentrations under aerobic condition(P<0.01). 8. IVDMD values of wilted(WT), beet pulp added(BP), wilted and LAB inoculated(W1) and beet pulp added and LAB inoculated(B1) silages were higher than direct-cut treatmentPC) as 84.7, 84.7, 84.4 and 83.0%, respectively on 0 day. But during 7-day period of aerobic exposure, a great decreae in IVDMD of WL was showed(P<0.01). The experimental results indicate that wilted silage(WT) could be recommended as the most effective treatment for reducing efluent and increasing quality and feed value of rye silage without deteriorating aerobic stability more than direct cut treatment(DC). Additionally, under unfavorable weather condition beet pulp added and LAB inoculated treatment(T3L) might be the possible alternative for successful ensiling of forage rye.

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Comparisons of the Forage Productivity and Mineral Contents Between the Newly Reclaimed and Arable Pasture Soils II. Changes in the soil properties, vigour at early growth, yields, and mineral nutrients of forages as affected by lime application (신개간 및 기경작 초지토양간 목초의 생산성과 무기양분 함량 비교 II. 석회 시용에 따른 토양특성, 목초의 초기생육, 수량 및 무기양분 함량 변화)

  • Jung, Yeun-Kyu
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.4
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    • pp.279-286
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    • 2002
  • This pot experiment was conducted to find out the forage productivity and contents of mineral nutrients in an orchardgrass sward affected by the compost lime, and phosphorus applications on the newly reclaimed and arable pasture soils. This 2nd part was concerned with the effect of lime application(L$_{0}$=control, L$_1$=279, L$_2$=54g CaCO$_3$/pot). The results obtained are summarized as follows: 1. There were considerable differences between the newly reclaimed and arable pasture soils in the soil chemical properties as follows in general; poor/common in the newly reclaimed soil longrightarrow and common/good in the arable pasture soil in terms of soil fertility standard. 2. It was recognized that the lime application was resulted in somewhat decrease of the concentrations of available P$_2$O$_{5}$ and increase of P$_2$O$_{5}$ absorbtion coefficient in the newly reclaimed soil, whereas these negative effects were positively enhanced by both lime and compost applications. The concentrations of exchangeable Ca and mg in both soils were increased by lime application. 3. The vigour at early growth stage of orchardgrass was better in the arable pasture soil than newly reclaimed soil. It was worsened by the heavy lime application(L$_2$), but this negative effect was positively enhanced by both lime and compost application. 4. Compared with the non-lime application, the forage yields of L$_1$ treatment were increased in both soils by 15~18%. whereas it of L$_2$ treatment was similar or lower than it of L$_1$. 5. The concentrations of Ca, mg, and P in orchardgrass were relatively higher in the arable pasture soil than the newly reclaimed soil, whereas there it was no difference in the K concentration. It was recognized that the concentrations of mg and Ca in orchardgrass were higher in the L$_1$ treatment than the Lo treatment in both soils, whereas there were no difference in it between L$_1$ and L$_2$ treatments.ments.

Studies on the Associative N2-fixing Microbes in Crop Root Systems -I. Associative Nitrogen Fixation by Azospirillum Isolated from Weed Roots grown Rice Paddy Field and its Distribution (작물근계(作物根系) 협생질소(協生窒素) 고정미생물(固定微生物)에 관(關)한 연구(硏究) -I보(報) : 논 토양(土壤) 잡초근계(雜草根系) 협생질소(協生窒素) 고정미생물(固定微生物)의 분포(分布)와 질소고정(窒素固定))

  • Ryu, Jin-Chang;Lee, Sang-Kyu;Park, Jun-Kyu;Ko, Jae-Young;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.17 no.4
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    • pp.415-427
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    • 1984
  • This experiment was carried out to investigate the distribution of associative nitrogen fixers, Azospirillum spp. and their nitrogenase activities measured by ARA in the rhizosphers of rice, soybean, and weed grown in the rice paddy field at ear formation stage of rice. Nitrogenase activities produced by Azospirillum spp. enriched from histophere ranged from 16 to 53 n mole/tube/hr.. High nitrogen fixing activities more than 30 n mole/tube/hr. were observed in the histophere of the Echinochloa crus-galli L., Finbristylis miliacea L., and Monochoria vaginalis var.. Nitrogen fixing activities of Azospirillum spp. obtained from single colonies which originated from the rhizoplane of rice (pot-kwang var.), Finbris tylis miliacea L., Monochoria vaginaliz var., Glycine max L. were higher over 100 n mole/tube/hr. than those histophere. Genus of Azopsirillum isoltated from roots of the Graminease (Oriza sativa L., $C_3$-plant, Echinochloa crus=galli L., $C_4$-plant, Cyperus difforuis L.. $C_4$-plant), and Aeschynomene indica L. (Leguminosae, $C_3$-plant) was identified as A. brasilense. However, both strains, A. lipoferum and A. brasilense ($nir^-$ or $nir^+$ strain) were isolated from other plant roots, Both $nir^-$ and $nir^+$ strains of A. brasilense were associated with the same host plant.

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