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N-Acyl-Homoserine Lactone Quorum Sensing Switch from Acidogenesis to Solventogenesis during the Fermentation Process in Serratia marcescens MG1

  • Jin, Wensong;Lin, Hui;Gao, Huifang;Guo, Zewang;Li, Jiahuan;Xu, Quanming;Sun, Shujing;Hu, Kaihui;Lee, Jung-Kul;Zhang, Liaoyuan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.596-606
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    • 2019
  • N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ${\Delta}swrI$ with or without supplementing exogenous N-hexanoyl-L-homoserine lactone ($C_6-HSL$) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing $C_6-HSL$. Furthermore, fermentation product analysis indicated that ${\Delta}swrI$ could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ${\Delta}swrI$ appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, ${\alpha}$-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.

Improvement of Cell Viability Using a Rho-associated Protein Kinase (ROCK) Inhibitor in Human Dental Papilla derived Single-induced Pluripotent Stem Cells (ROCK 억제제를 통한 사람 치유두 조직 유래 단일 사람 유도만능줄기세포의 생존성 향상)

  • Shim, Yoo-Jin;Kang, Young-Hoon;Kim, Hyeon-Ji;Kim, Mi-Jeong;Lee, Hyeon-Jeong;Son, Young-Bum;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.29 no.8
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    • pp.895-903
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    • 2019
  • The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.

Ecological Studies on the Inhavitation of Artificial Nests by Wild Birds -Especially in Gyeongnam Province- (인공소상가설(人工巢箱架設)에 의(依)한 야생조류(野生鳥類)의 서식생태(棲息生態)에 관(關)한 연구(硏究) -경남지역(慶南地域)을 중심(中心)으로-)

  • Choi, Jai-Sik;Kim, Jai-Saing
    • Journal of Korean Society of Forest Science
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    • v.76 no.2
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    • pp.109-118
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    • 1987
  • The purpose of this study was to investigate the inhabiting ecology of wild birds using artificial nests in Gyeongnam province. The sort of birds, the utilization of artificial nests by the materials, by the sites and by the forest type, and other inhabiting ecology of birds were investigated from 1984 to 1986. The results obtained were as follows; 1. Wild birds in surveyed areas were classified into 20 families and 35 species. Among them the birds inhabiting the artificial nests were 6 species and those using them most frequently were Parus ater (17.2%), P. major (16.7%) and P. palustris (12.2%). 2. The artificial nests installed on the top and at the toot of the mountain were chiefly utilized by P. ater and P. major, but the artificial nests in park area were mostly used by Passer montanus and Sturnus cineraceus. 3. Sixty-one percent of the artificial nests were utilized by the birds; 45.5% were used for breeding and 15.6% were inhabitation only. 4. When different nest materials and shapes were tested, mixed nests (sawdust and cement) were used most frequently, and C type. (Diamond shape) of wood nests was used more frequently compared with the A (Roof shape) and B (Box shape) types of wood nests. Meanwhile, the birds prefered the 3cm hole size (81.1% of utilization) to 4cm of hole (57.8%) and 5cm of hole (24.4%). 5. The artificial nests constructed in mixed forest were more frequently inhabited (73.3%) by the birds than coniferous forest (68.3%) and broad-leaved forest (63.3%). 6. The period of egg-laying in Parus species was from late April to early May, while Eophona migratoria, Lanius bucephalus, Motacilla alba and Passer montanus laid eggs generally in the middle of May. 7. Parus species, Passer montanus and Motacilla alba laid one egg every day for 5-10days, but Eopphona migratoria laied 4 eggs in 7 days and Lanius bucephalus laid 5 eggs in 4 days. The incubation period of Parus species was 16-18 days, while the others were about 11 to 14 days. 8. Thirteen days after the hatch of Parus major, chicks reached 13.9g of body weight, 72.0mm of wing-length and 20.9mm of tarsus length. P. montanus chicks reached 20.3g of weight, 66.2mm of wing-length and 20.2mm of tarsus length during the same period. 9. Food item of feeding chicks almost consisted of creatures (98.2%) with following composition; 55.2% of insect larvae, 37.2% adult insects, 2.8% of pupae and 2.8% of spiders. Vegetable items were 2% only. Among those creature food items, 95.2% were insect pests to forest.

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Usefulness of Pulsatile Flow Aortic Aneurysm Phantoms for Stent-graft Placement (스텐트그라프트 장치술을 위한 대동맥류 혈류 팬텀의 유용성)

  • Kim, Tae-Hyung;Ko, Gi-Young;Song, Ho-Young;Park, In-Kook;Shin, Ji-Hoon;Lim, Jin-Oh;Kim, Jin-Hyoung;Choi, Eu-Gene K.
    • Journal of radiological science and technology
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    • v.30 no.3
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    • pp.205-212
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    • 2007
  • To evaluate the feasibility and efficacy of a pulsatile aortic aneurysm phantoms for in-vitro study. The phantoms consisted of a pulsating motor part(heart part) and an aortic aneurysm part, which mimicked true physiologic conditions. The heart part was created from a high-pressured water pump and a pulsatile flow solenoid valve for the simulation of aortic flow. The aortic aneurysm part was manufactured from paper clay, which was placed inside a acrylic plastic square box, where liquid silicone was poured. After the silicone was formed, the clay was removed, and a silicone tube was used to connect the heart and aneurysm part. We measured the change in pressure as related to the opening time(pulse rate, Kruskal-Wallis method) and pressure before and after the stent-graft implantation(n = 5, Wilcoxon's signed ranks test). The changes in blood pressures according to pulse rate were all statistically significant(p<0.05). The systolic/diastolic pressures at the proximal aorta, the aortic aneurysm, and the distal aorta of the model were $157.80{\pm}1.92/130.20{\pm}1.92$, $159.40{\pm}1.14/134.00{\pm}2.92$, and $147.20{\pm}1.480/129.60{\pm}2.70\;mmHg$, respectively, when the pulse rate was 0.5 beat/second. The pressures changed to $161.40{\pm}1.34/90.20{\pm}1.64$, $175.00{\pm}1.58/93.00{\pm}1.58$, and $176.80{\pm}1.48/90.80{\pm}1.92\;mmHg$, respectively, when the pulse rate was 1.0 beat/second, and $159.40{\pm}1.82/127.20{\pm}1.48$, $166.60{\pm}1.67/138.00{\pm}1.87$, and $161.00{\pm}1.22/135.40{\pm}1.67\;mmHg$, respectively, when it was 1.5 beat/second. When pulse rate was set at 1.0 beat/second, the pressures were $143.60{\pm}1.67/90.20{\pm}1.64$, $147.20{\pm}1.92/84.60{\pm}1.82$, and $137.40{\pm}1.52/88.80{\pm}1.64\;mmHg$ after stent-graft implantation. The changes of pressure before and after stent-graft implantation were statistically significant(p<0.05) except the diastolic pressures at the proximal(p =1.00) and distal aorta(p=0.157). The aortic aneurysm phantoms seems to be useful for the evaluation of the efficacy of stent-graft before animal or clinical studies because of its easy reproducibility and ability to display a wide range of pressures.

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Effects of CDP-Choline, Aminoguanidine and Difluoromethylornithine on the ECS-induced Impairment of Active Conditioned Response Retention (백서의 조건회피반응-유지에 대한 경련성 전기충격의 저해작용에 미치는 CDP-Choline, Aminoguanidine, 및 Difluoromethylornithine의 영향에 관한 연구 : 뇌내 Acetylcholine과 Polyamine 함량-변동에 연관하여)

  • Kim, Hyung-Gun;Kim, Chang-Hyun;Choi, Sang-Hyun;Ihm, Suk-Young;Lee, Min-Soo;Chun, Boe-Gwun
    • The Korean Journal of Pharmacology
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    • v.28 no.2
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    • pp.115-128
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    • 1992
  • The training of male wistar rats for active conditioned response (ACR) was performed by one daily training session of 30 consecutive trials for 10 successive days using a two-way shuttle box, and the rats that showed 10 or more ACRs on the last day were treated for further 10 days with electroconvulsive shock (ECS : 50 mA, 0.5 msec; 100 Hz; 1.5 sec) and the following compounds. On the 20th day, all the rats were tested for the ACR rention. The ECS regimens were one ECS per day for 10 days with one day interval $(5{\times}ECS)$, one ECS at 3 hrs (ECS-3h), and one ECS at 24 hrs (ECS-24h), respectively, before the ACR retention test. And CDP-choline (cc: 250 mg/kg), spermine (SM: 10 mg/kg), ${\alpha}-difluoromethylornithine$ (DO: 250 mg/kg), or aminoguanidine (AG: 100 mg/kg) was administered by one daily i.p. injection for 10 days. The ACR number $(13.7{\pm}1.0)$ obtained on the last training day was increased by 37.23% on the 20th day in the control rats. And the ACR increase was significantly suppressed by 5-ECS, ECS-3h, CC, or SM but was little affected by ECS-24h, DO, or AG. However, the 5-ECS induced impairment of ACR retention was significantly suppressed by AG, SM, and CC in the order of potency but was little affected by DFMO. And the ECS-3h induced impairment was moderately worsened by SM or AG. The acetylcholine (ACh) of the rat hypothalamus (HT), hippocampus (HC), and entorhinal cortex (EC) was markedly increased by CC and moderately increased by SM, but little affected by ECS-3h, ECS-24h, DO, or AG. But $5{\times}ECS$ slightly increased the ACh content. The brain putrescine (Pt) content was significantly increased by AG and little affected by CC, SM, or DO. But the $5{\times}ECS$ markedy decreased the brain Pt content, and the decrease was significantly suppressed by CC, SM, or AG. CC induced the marked increases of the spermidine (Sd) and spermine (Sm) contents of all the areas. SM increased the Sd contents of all the areas and the EC-Sm content. DO decreased the brain Sd and Sm contents. And AG increased the HT-Sd content and the Sm contents of all the brain areas. The $5{\times}ECS$ induced decrease of the HC-Sm content was suppressed by CC, SM and AG. These results suggest that the improving effect of aminoguanidine on the $5{\times}ECS$ induced impairment of ACR retention may be ascribed in part to its activity as a diamine oxidase inhibitor.

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Determination of the Optimum Sampling Area for the Benthic Community Study of the Songdo Tidal Flat and Youngil Bay Subtidal Sediment (송도 갯벌과 영일만 조하대 저서동물의 군집조사를 위한 적정 채집면적의 결정)

  • Koh, Chul-Hwan;Kang, Seong-Gil;Lee, Chang-Bok
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.1
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    • pp.63-70
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    • 1999
  • The optimum sampling area which can be applied to the benthic community study is estimated from large survey data in the Songdo tidal flat and subtidal zone of Youngil Bay, Korea. A total of 250 samples by 0.02 $m^2$ box corer for the benthic fauna in Songdo tidal flat and 50 samples by 0.1 $m^2$ van Veen grab in Youngil Bay were taken from the total sampling area of 5 $m^2$. It was assumed that the sampling area could contain sufficient information on sediment fauna, if cumulative number of species, ecological indices, and similarity index by cluster analysis reflect the similarity level of 75% to those found at total sampling area (5 $m^2$). A total of 56 and 60 species occurred from Songdo tidal flat and Youngil Bay, respectively. The cumulative curve of the species number ($N_{sp}$) as a function of the sampling area (A in $m^2$ ) was fitted as $N_{sp}=37.379A^{0.257}$ ($r^2=0.99$) for intertidal fauna and $N_{sp}=40.895A^{0.257}$ ($r^2=0.98$) for subtidal fauna. Based on these curves and 75% of similarity to the total sampling area (5 $m^2$), the optimum sampling area was proposed as 1.6 $m^2$ for the intertidal and 1.5 $m^2$ for the subtidal fauna. Ecological indices (species diversity, richness, evenness and dominance indices) were again calculated on the basis of species composition in differently simulated sample sizes. Changes in ecological indices with these sample sizes indicated that samplings could be done by collecting fauna from < 0.5 $m^2$-1.5 $m^2$ on the Songdo tidal flat and from < 0.5 $m^2$-1.2 $m^2$ in Youngil Bay. Changes in similarity level of all units of each simulated sample size showed that sampling area of 0.3 $m^2$ (Songdo tidal flat) and 0.6 $m^2$ (Youngil Bay) should be taken to obtain a similarity level of 75%. In conclusion, sampling area which was determined by cumulative number of species, ecological indices and similarity index by cluster analysis could be determined as 1.5 $m^2$ (0.02 $m^2$ box corer, n=75) for Songdo tidal flat and 1.2 $m^2$ (0.1 $m^2$ van Veen grab, n=12) for Youngil Bay. If these sampling areas could be covered in the field survey, population densities of seven dominant species comprising 68% of the total faunal abundance occurring on Songdo tidal flat and six species comprising 90% in Youngil Bay can be estimated at the precision level of P=0.2.

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Processing of Cheonggukjang using soybean powder (콩 분말을 이용한 청국장 제조 기술 개발)

  • Jeong, Yeon-Shin;Dhakal, Krishna Hari;Hwang, Young-Hyun
    • Current Research on Agriculture and Life Sciences
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    • v.29
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    • pp.43-53
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    • 2011
  • The possibility of making Cheongukjang by the use of soy-powder and pill type re-made from soy-powder was investigated. Some of experimental results obtained are summarized as follows; 1. Regardless of three different types for the source material of Cheonggukjang, soybean seed, soy-power, and pill, the dry weight of fermented Cheonggukjang showed continuously decreasing trends along with the time of fermentation applied. 2. In all type of source materials, the length of viscous substance during fermentation was increased along with the time of fermentation, and the rate of elongation was much reduced after 48hr of fermentation. Out of four soybean varieties tested, Taekwangkong produced longest viscous substance fermented in the type of soybean seed. No viscous substance was formed when the depth of soy-powder in the fermentation box was shallow, one centimeter. 3. Not much difference was observed in the number of microbes, Bacillus licheniformis B1, in all soybean varieties. The number was proportionally increased in the type of powder but it increased rapidly from 12 hour to 24 hour with low rate of increase thereafter in the pill type. 4. Along with the time of fermentation in all types, the color of Cheonggukjang changed from yellow to dull. At the same time, the tone of color and chroma changed into reddish and yellow, respectively. 5. Along with the time of fermentation in all types, pH of Cheonggukjang changed in alkali. 6. Along with the time of fermentation, the content of isoflavone in Cheonggukjang increased by 48hr but decreased thereafter. 7. In general, the quality of Cheonggukjang fermented in types of soy-powder and pill re-made from soy-powder was lower than that of soybean seed. More study is seemed to be needed to produce high quality Cheonggukjang by the use of soy-power.

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Proinflammatory Effects of High Mobility Group B1 (HMGB1) Versus LPS and the Mechanism of IL-8 Promoter Stimulation by HMGB1 (High mobility group B1(HMGB1)과 LPS의 염증유발효과 차이의 비교 및 HMGB1에 의한 IL-8 promoter 자극 기전의 규명)

  • Jeon, Eun Ju;Kwak, Hee Won;Song, Ju Han;Lee, Young Woo;Chung, Jae Woo;Choi, Jae Chul;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.4
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    • pp.299-307
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    • 2007
  • Background: High mobility group box 1 (HMGB1) is a novel, late mediator of inflammation. This study compared the pro-inflammatory effects of LPS and HMGB1. The transcriptional factors that play an important role in mediating the HMGB1-induced stimulation of IL-8 were also evaluated. Methods: RAW264.7 cells were stimulated with either LPS (100 ng/ml) or HMGB1 (500 ng/ml). The $TNF-{\alpha}$, MIP-2 and $IL-1{\beta}$ levels in the supernatant were evaluated by ELISA at 0, 2, 4, 8, 12 and 24h after stimulation. An acute lung injury was induced by an injection of LPS (5 mg/kg) or HMGB1 (2.5 mg/kg) into the peritoneum of the Balb/c mice. The lung cytokines and MPO activity were measured at 4h (for LPS) or 24h (for HMGB1) after the injection. The transcriptional factor binding sites for NF-IL6, $NF-{\kappa}B$ and AP-1 in the IL-8 promoter region were artificially mutated. Each mutant was ligated with pIL-6luc and transfected into the RAW264.7 cells. One hour after stimulation with HMGB1 (500 ng/ml), the cell lysate was analyzed for the luciferase activity. Results: The expression of MIP-2, which peaked at 8h with LPS stimulation, increased sequentially until 24h after HMGB1 stimulation. An intraperitoneal injection of HMGB1, which induced a minimal increased in $IL-1{\beta}$ expression, provoked the accumulation of neutrophils the lung. A mutation of AP-1 as well as $NF-{\kappa}B$ in the IL-8 promoter region resulted in a lower luciferase activity after HMGB1 stimulation. Conclusion: The proinflammatory effects of HMGB1, particularly on IL-8, are mediated by both $NF-{\kappa}B$ and AP-1.

Processing Conditions and Quality Stability of Frozen Seasoned Sardine Meat during Frozen Storage (냉동 정어리 조미육의 가공 및 저장중의 품질안정성)

  • LEE Eung-Ho;OH Kwang-Soo;AHN Chang-Bum;LEE Tae-Hun;CHUNG Young-Hoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.3
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    • pp.191-201
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    • 1987
  • Seasoned sardine meat was prepared to extend the use of sardine for human consumption, and processing conditions and storage stability of frozen seasoned sardine meat were studied during storage at $-20^{\circ}C$. The fish was beheaded, gutted and cleaned in a washing tank. The washed fish was then put through a belt-drum type meat separator which separates the flesh iron the bone and skin. Mechanically deboned fish meat was mixed with $20.6\%$ emulsion curd, $0.5\%$ table salt, $2.0\%$ sugar, $0.4\%$ sodium bicarbonate, $0.2\%$ polyphosphate, $0.1\%$ monosodium glutamate, $0.3\%$ onion powder, $0.1\%$ garlic powder, $0.1\%$ ginger powder, $3.0\%$ soybean protein and $0.1\%$. In sodium erythorbate. This seasoned sardine meat was frozen with contact freezer, packed in a carton box and then stored at $-20^{\circ}C$. The pH, volatile basic nitrogen, viable cell counts, peroxide value, carbonyl value, thiobarbituric acid value, taste compounds, fatty acid composition, salt extractable nitrogen, drip, texture, and color values of the products were determined during frozen storage. The results showed that lipid content in products could be controlled by using emulsion curd, and flavor and texture could be improved by adding spices and soybean protein, and lipid oxidation could be retarded by $0.1\%$ sodium erythorbate. Judging from the results of chemical experiments and sensory evaluation, the products can be preserved in a good quality for 120 days during frozen storage.

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Silicon Uptake Level of Six Potted Plants from a Potassium Silicate-supplemented Hydroponic Solution (규산칼륨 첨가 양액으로부터 6가지 분식물의 규소 흡수도)

  • Son, Moon Sook;Song, Ju Yeon;Lim, Mi Young;Sivanesan, Iyyakkannu;Kim, Gui Soon;Jeong, Byoung Ryong
    • Horticultural Science & Technology
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    • v.31 no.2
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    • pp.153-158
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    • 2013
  • This research was carried out to investigate silicon (Si) uptake levels by six potted plant species from a nutrient solution supplemented with $K_2SiO_3$. Uniform rooted plants of Dendranthema grandiflorum Ramat., Spathiphyllum patinii N.E. BR., Kalanchoe blossfeldiana, Hedera helix L., Dianthus caryophyllus L., and Euphorbia pulcherrima Willd. were grown in 350 mL boxes, one plant per box, containing a nutrient solution supplemented with either 0, 2.7, or 5.4 mM Si as $K_2SiO_3$. The nutrient solution in each container was adjusted to EC $1.5mS{\cdot}cm^{-1}$ and pH 5.6. The solution in each container was aerated by an 1 m-long polyethylene tube, all connected to a vacuum pump. After 15 days of cultivation in a glasshouse Si contents in the roots and shoots were measured using the colorimetric molybdate method and amount of remaining Si in the nutrient solution was measured using the ICP-AES to calculate the amount of absorption. A simple regression analysis was performed to observe the changes in Si contents in the roots and shoots as affected by concentration of Si supplied to the solution. Among the six species tested, carnation had the greatest and poinsettia the lowest tissue levels of Si concentration in the root, whereas carnation had the greatest and kalanchoe the lowest tissue levels of Si concentration in the shoot. Based on the Si content in the whole plant, Si uptake levels by poinsettia, kalanchoe, and chrysanthemum were low, whereas those by spathiphyllum were intermediate, and those of English ivy and carnation were high. These results indicated that the uptake level of Si by the plant vary depending on plant species.