• Title/Summary/Keyword: Fluorescence effect

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Antimicrobial Effects against Food-Borne Pathogens and Antioxidant Activity of Rhododendron brachycarpum Extract (만병초 추출물의 식중독 유발균에 대한 항균효과 및 항산화활성)

  • Choi, Moo-Young;Rhim, Tae-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1353-1360
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    • 2011
  • This study was performed to investigate the antimicrobial effects against food-borne pathogens and antioxidant activity of Rhododendron brachycarpum ethanol-extract. The antimicrobial activity of the extract was determined using a paper disc-diffusion method, and the diameter of the clear zone was measured. The diameter of the clear zone in the presence of 10 mg of extract was maximal against Bacillus cereus among the three tested Gram-positive bacteria and against Escherichia coli O157:H7 among the five tested Gram-negative bacteria. Analysis of the minimum inhibitory concentration (MIC) showed that the extract exhibited a similar efficacy as that of sorbic acid, a well-known chemical preservative. The growth inhibitory effects of the extract at concentrations of 250, 500, 1,000, and 2,000 mg/L on food-borne pathogens were determined against Staphylococcus aureus, Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7. Growth of the microorganisms was not affected by the extract at concentrations up to 250 mg/L, but it was significantly (p<0.05) inhibited by the extract at concentrations higher than 1,000 mg/L. The antioxidant effects of the extract were examined via measurement of DPPH radical scavenging activity, inhibition of reactive oxygen species (ROS) generation using fluorescent dichlorofluorescien (DCF) assay, and prevention of peroxyl radical- and hydroxyl radical-induced supercoiled DNA breakage. The $IC_{50}$ of the extract for DPPH radical scavenging activity was about half that of ${\alpha}$-tocopherol, which was used as a positive control. DCF fluorescence intensity decreased as the concentration of the extract increased, demonstrating that ROS generation was inhibited in a concentration-dependent manner. The ROS inhibitory effect of the extract was higher than that of ascorbic acid. The extract prevented supercoiled DNA strand breakage induced by peroxyl radical and hydroxyl radical. Thus, the results of the present study demonstrate that the extract exhibits antimicrobial effects against food-borne pathogens as well as potent antioxidant capacity, suggesting that R. brachycarpum could be used as a natural antibacterial agent and effective antioxidant in food.

Effect of Different Fertilization on Physiological Characteristics and Growth Performances of Eucalyptus pellita and Acacia mangium in a Container Nursery System (시비처리가 Eucalyptus pellita와 Acacia mangium 용기묘의 생리 및 생장 특성에 미치는 영향)

  • Cho, Min-Seok;Lee, Soo-Won;Bae, Jong-Hyang;Park, Gwan-Soo
    • Journal of Bio-Environment Control
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    • v.20 no.2
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    • pp.123-133
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    • 2011
  • The objective of this study was to find optimal nutrient condition of container seedling production of two tropical species for high seedling quality. This study was conducted to investigate photosynthesis, chlorophyll fluorescence, chlorophyll contents, and growth performances of container seedlings of Eucalyptus pellita and Acacia mangium growing under four different fertilization treatments (Con., $0.5\;g{\cdot}l^{-1}$, $1.0\;g{\cdot}l^{-1}$, and $2.0\;g{\cdot}l^{-1}$ fertilization). E. pellita showed outstanding photosynthetic capacity, photochemical efficiency, and chlorophyll contents at $1.0\;g{\cdot}l^{-1}$ fertilization. Meanwhile, E. pellita showed the highest photosynthetic capacity, photochemical efficiency, and chlorophyll contents at $2.0\;g{\cdot}l^{-1}$ fertilization, as fertilization rate were increased, those of A. mangium increased. Like physiological characteristics, Both E. pellita at $1.0\;g{\cdot}l^{-1}$ fertilization and A. mangium at $2.0\;g{\cdot}l^{-1}$ fertilization were higher root collar diameter, height, biomass, and seedling quality index than other treatments. These results showed that E. pellita at $1\;g{\cdot}l^{-1}$ fertilization and A. mangium at $2.0\;g{\cdot}l^{-1}$ fertilization is optimal nutrient condition, respectively. Moreover, fertilization rate controlling is very important for growth and seedling quality of container seedling.

Effects of microorganism density and mushroom yields according to the sterilization of casing soils at the cultivation of button mushrooms (복토살균 조건에 따른 양송이 재배과정별 복토내 미생물 밀도 및 수량 특성)

  • Lee, Chan-Jung;Yoo, Young-Mi;Jhune, Chang-Sung;Cheong, Jong-Chun;Moon, Ji-Won;Kong, Won-Sik;Suh, Jang-Sun;Kim, Yong-Gyun;Lee, Byung-Eui;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.220-225
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    • 2014
  • This study was conducted to set the proper sterilization standards of casing soil for the stable production of button mushroom(Agaricus bisporus) from mushroom disease that occurs in infection of casing soil material. Changes of aerobic bacteria are increased as the longer grow-out period and sharply increased after second flushes. Fluorescence Psuedomonas showed high density at high sterilization temperature and $100^{\circ}C$ treatment has extremely high density at 30 min and 60 min in casing 22 days. Density of thermophilic actinomyces is sharply increase from casing with soil and the highest density at 22 days of casing and rapidly decrease after first flushes. Sterilizing temperature of casing soil affects quality and quantity of button mushroom. Treatment of 60 min, 90 min at $80^{\circ}C$ and 30 min at $100^{\circ}C$ produced the highest mushroom yields, especially mushrooms yields of A grads were the highest at treatment of 90 min at $80^{\circ}C$. Treatment of 60min at $100^{\circ}C$ products many yields, however, this treatment has low economic feasibility for its yields. Sterilizing temperature of casing soil has an effect on generating diseases and insect pests. Treatment of 60 min, 90 min at $80^{\circ}C$ and 30 min $100^{\circ}C$ showed lower incidence than the other treatment. Although treatment of 30 min at $100^{\circ}C$ causes low diseases and mushroom fly damage, it has low mushroom yields. Furthermore, although treatment of 60 min at $100^{\circ}C$ has high mushroom yields, it causes high diseases and mushroom fly damage. Therefore the best conditions for the sterilization of casing soils was 60 min and 90 min at $80^{\circ}C$.

Estimation of POC Export Fluxes Using 234Th/238U Disequilibria in the Amundsen Sea, Antarctica; Preliminary Result (남극 아문젠해에서 234Th/238U 비평형법을 사용한 유광대에서 심층으로의 입자상 유기탄소 침강플럭스 추정; 예비결과)

  • Kim, Mi Seon;Choi, Man Sik;Lee, Sang Heon;Lee, Sang Hoon;Rhee, Tae Siek;Hahm, Doshik
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.19 no.2
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    • pp.109-124
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    • 2014
  • In order to understand the carbon cycle in the Amundsen Sea of the Southern Ocean, the export fluxes of particulate organic carbon from the euphotic zone to deep water estimated using ${\psi}$/${\psi}$ disequilibrium method. Seawaters in 14 water columns were collected during February and March 2012, and analyzed for total and dissolved ${\psi}$, and particulate organic carbon. Total ${\psi}$ activities in the water column showed deficiency and excess relative to those of ${\psi}$ depending on the water depth. Deficiency of total ${\psi}$ in the euphotic zone showed mirror images both with chlorophyll-a and fluorescence, and was consistent with the loss of nitrate, which indicated the effect of biological activity. In addition, deficiency of total ${\psi}$ from deep water was associated with the increase of total dissolvable Fe/Mn concentration. Excess total ${\psi}$ activity presented below the euphotic zone might be related to particulate ${\psi}$ concentrated in this water depth. Mean export flux of ${\psi}$ estimated using the steady state model was $867{\pm}246dpmm^{-2}day^{-1}$. Mean export flux of particulate organic carbon, which were estimated by the product of total ${\psi}$ flux and ratio of POC/${\psi}$ ($7.08{\pm}4.27{\mu}molCdpm^{-1}$) in the sinking particles, was $5.9{\pm}3.9mmolCm^{-2}day^{-1}$. These fluxes were similar levels to those in the Weddell Sea during February and March 2008. Export ratios (ThE) relative to the primary production in the euphotic zone were in the range of 3-54% (av. 28%).

Analysis of PAHs (polycyclic aromatic hydrocarbons) in Ground Coffee Using GC-tandem Mass Spectrometry and Estimation of Daily Dose (GC-tandem mass spectrometry를 이용한 분쇄원두커피 중 PAHs(polycyclic aromatic hydrocarbons) 분석법 연구 및 인체노출량 평가)

  • Jung, So-Young;Park, Ju-Sung;Son, Yeo-Joon;Choi, Su-Jeong;Lee, Yun-Jeong;Kim, Mi-Sun;Park, So-Hyun;Lee, Sang-Me;Chae, Young-Zoo
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.544-552
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    • 2011
  • The purpose of this study was to develop an analytical method for determining 15 polycyclic aromatic hydrocarbons (PAHs) of EU priority using gas chromatography (GC)-tandem mass spectrometry (MS). The PAHs in ground coffee were analyzed after being extracted using methods such as saponification-liquid-liquid extraction, Soxhlet extraction, and solid-liquid extraction. The solid-liquid extraction method showed the greatest repeatability and most efficient reduction of the matrix effect. GC-tandem MS for the quantification of the 15 PAHs showed better resolution and lower limit of detections (LODs) than GC-MS-selected ion monitoring (SIM) and high performance liquid chromatography with fluorescence detector. LODs of this method for the ground coffee types were 0.002-0.1 ${\mu}g/kg$ and limit of quantifications (LOQs) were 0.006-0.2 ${\mu}g/kg$ The recoveries ranged from 52.6 to 93.3%. Forty-six commercial types of ground coffee were analyzed to determine their PAHs contamination levels. PAHs concentration ranged from ND to 5.988 ${\mu}g/kg$. This study was conducted with toxicity equivalence factors, the U.S. EPA recommendation to identify dietary risks for PAHs in different types of coffee. The estimated average daily dose of PAHs was $5.24{\times}10^{-8}$ mg/kg body weight/day.

β-elemene Induces Caspase-dependent Apoptosis in Human Glioma Cells in vitro through the Upregulation of Bax and Fas/FasL and Downregulation of Bcl-2

  • Li, Chen-Long;Chang, Liang;Guo, Lin;Zhao, Dan;Liu, Hui-Bin;Wang, Qiu-Shi;Zhang, Ping;Du, Wen-Zhong;Liu, Xing;Zhang, Hai-Tao;Liu, Yang;Zhang, Yao;Xie, Jing-Hong;Ming, Jian-Guang;Cui, Yu-Qiong;Sun, Ying;Zhang, Zhi-Ren;Jiang, Chuan-Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10407-10412
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    • 2015
  • Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

The study on cytotoxicity of cytokines produced by the activated human NKT cells on neuroblastoma (활성화된 자연살상 T 세포(NKT)에서 생성된 사이토카인에 의한 신경모세포종의 세포독성에 관한 연구)

  • Cho, Jin Young;Yoon, Young Wook;Yoon, Hyang Suk;Kim, Jong Duk;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.4
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    • pp.439-445
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    • 2006
  • Purpose : ${\alpha}$-Galactosylceramide (${\alpha}$-GalCer)-stimulated human $V{\alpha}24$ natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d - negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/${\alpha}$ GalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. Methods : Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. Results : The activated NKT cells secreted high levels of IL-2, INF-${\gamma}$, TNF-${\alpha}$. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-${\alpha}$ and anti-IFN-${\gamma}$ neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. Conclusion : IFN-${\gamma}$ and TNF-${\alpha}$ produced by NKT cells could exert synergistically direct antitumor activity through apoptosis on some NB cell lines.

Effects of Contrast Improvement on High Voltage Rectification Type of X-ray Diagnostic Apparatus (X선 진단장치의 고압정류방식이 대조도 향상에 미치는 영향)

  • Lee, Hoo-Min;Yoon, Joon;Kim, Hyun-Ju
    • Journal of radiological science and technology
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    • v.37 no.3
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    • pp.187-193
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    • 2014
  • The purpose of this study was to analyze the effect on the selectivity on of high-voltage rectification device that measured the performance of the grid, and the contrast improvement ability (K factor) by measuring the scattered radiation content of the transmitted X-rays. The scattered radiation generated when the X-ray flux comes from the diagnostic X-ray generator that passes through an object. Targeting four different rectifications of X-ray generators, the mean value of the tube voltage and the tube current was measured in order to maximize the accuracy of the generating power dose within the same exposure condition. Using fluorescence meter, the content of the scattered rays that are transmitted through the acrylic was measured depending on the grid usage. When grid is not used, the content of the scattered rays was the lowest (34.158%) with the single-phase rectifier, was increased with the inverter rectifier (37.043%) and the three-phase 24-peak rectification method (37.447%). The difference of the scattered radiation content of each device was significant from the lowest 0.404% to the highest 3.289% while using 8:1 grid, the content of the scattered ray was the lowest with the single content of the scattered ray was the lowest with the single-phase rectifier (18.258%), was increased with the rectifier (25.502%) and the 24-peaks rectification (24.217%). Furthermore, there was difference up to content 7.244% to the lowest content 1.285% within three-phase 24-peaks rectification, inverter rectifications, and single-phase rectifier depending on the selectivity of the grid. Drawn from the statistical analysis, there was a similar relationship between the contrast improvement factor and the K factor. As a result, the grid selectivity and the contrast were increased within the single-phase rectifier rather than the constant voltage rectifier.

Study of Hedyotis Diffusa Methanol Extract on Anti-tumoral Effect and Mechanism (백화사설초(白花蛇舌草) 메탄올 추출물(抽出物)의 항종양(抗腫瘍) 효과(效果) 및 항암(抗癌) 기전(機轉)에 관(關)한 연구(硏究))

  • No, Hoon-Jeong;Moon, Gu;Moon, Seok-Jae;Won, Jin-Hee;Moon, Young-Ho;Park, Rae-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.6 no.1
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    • pp.81-97
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    • 2000
  • Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

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The Effect of IAA on $Colocasia$ $esculenta$'s Growth and Morphogenesis (IAA가 토란의 생장 및 형태 발생에 미치는 영향)

  • Kim, Young-Eun;Lee, Joon-Sang
    • Korean Journal of Environmental Biology
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    • v.29 no.2
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    • pp.92-97
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    • 2011
  • Morphological and physiological differences of $Colocasia$ $esculenta$ were investigated in the cultivation of hydorphonic and soil culture. $C.$ $esculenta$ grown in Hoa.+IAA (indole-acetic acid) showed higher growth activity representing 9%, 32%, 38% and 60% than those of the cultivation of vermiculate, Hoagland solution, soil and water, respectively. In case of $F_v/F_m$ ratio experiments, the value $F_v/F_m$ of $C.$ $esculenta$ cultivated in the water showed 0.55 after 6 weeks. $F_v/F_m$ values of $C.$ $esculenta$ cultivated in Hoagland+IAA, vermiculate and soil were between 0.84 and 0.80 indicating $F_v/F_m$ values were about 45% higher than that of $C.$ $esculenta$ cultivated in the water. Diffusion resistance was 45~35% lower in $C.$ $esculenta$ grown in Hoa.+IAA solution than that of $C.$ $esculenta$ grown in water only after 5 and 6 weeks. Therefore, the high standing levels of the growth rate, fluorescence activity and transpiration rate were Hoa.+IAA, vermiculate, Hoagland, soil and water. The distinct morphological differences of $C.$ $esculenta$ cultivated in hydorphonic and soil culture were the appearance of the seed corm and root hair. The development of seed corm was well established in soil culture but the corm in hydorphonic was slowly hydrolyzed and then disappeared. The fibrous root systems of hydorphonic were very well distinguishable compared with that in soil culture. Outstanding results of this experiment were appeared in $C.$ $esculenta$ which was cultivated in the field provided with enough mineral nutrition, organic fertilizers and compound fertilizers. The most height taros were almost 2m and the numbers of seed corm were 30~40 after 7 months.