• Title/Summary/Keyword: Plant Cell Culture

Search Result 622, Processing Time 0.032 seconds

Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.3
    • /
    • pp.147-152
    • /
    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

  • PDF

Biological Control of Crown Gall

  • Kerr, Allen;Biggs, John;Ophel, Kathy
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 1994.06a
    • /
    • pp.11-26
    • /
    • 1994
  • Crown gall of stonefruit and nut trees is one of the very few plant diseases subject to efficient biological control. The disease is caused by the soil-inhabiting bacteria Agrobacterium tumefaciens and Agrobacterium rhizogenes and the original control organism was a non-pathogenic isolate of A. rhizogenes strain K84. Control is achieved by dipping planting material in a cell suspension of strain K84 which specifically inhibits pathogenic strains containing a nopaline Ti plasmid. Because the agrocin 84-encoding plasmid (pAgK84) is conjugative, it can be transmitted from the control strain to pathogenic strains which, as a result, become immune to agrocin 84 and cannot be controlled. To prevent this happening, the transfer genes on pAgK84 were located and then largely eliminated by recombinant DNA technology. The resulting construct, strain K1026, is transfer deficient but controls crown gall just as effectively as does strain K84. Field data from Spain confirm that pAgK84 can transfer to pathogenic recipients from strain K84 but not from strain K1026. The latter has been registered in Australia as a pesticide and is the first genetically engineered organism in the world to be released fro commercial use. It is recommended as a replacement for strain K84 to prevent a breakdown in the effectiveness of biological control of crown gall. Several reports indicate that both strains K84 and K1026 sometimes control crown gall pathogens that are resistant to agrocin 84. A possible reason for this is that both strains produce a second antibiotic called 434 which inhibits growth of nearly all isolates of A. rhizogenes, both pathogens and non-pathogens. Crown gall of grapevine is caused by another species, Agrobacterium vitis. It is resistant to agrocin 84 and cannot be controlled by strains K84 or K1026. It is different from other crown gall pathogens in several characteristics, including the fact that, although a rhizosphere coloniser, its also lives systemically in the vascular tissue of grapevine. Pathogen free propagating material can be obtained from tissue culture or, less surely, by heat therapy of dormant cuttings. A number of laboratories are searching for a biocontrol strain that will prevent, or at least delay, reinfection. A non-pathogenic A. vitis strain F/25 from South Africa looks very promising in this regard.

  • PDF

Neuroprotective effects of antioxidant constituents isolated from Opuntia ficus-indica var. saboten Makino

  • K.J. Jung;Lee, E.H.;Kim, H.J.;Lee, J.Y.;Y.S. Song;Lee, Y.H.;J. Cho;Park, M.;Park, H.
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 2003.11a
    • /
    • pp.63-63
    • /
    • 2003
  • Opuntia ficus-indicavar. saboten Makino (Cactaceae) is a tropical or subtropical plant that has been widely used as folk medicine for the treatment of diabetes, asthma, burn, edema and gastritis. The purposes of the present study were to identify antioxidant constituents from fruits and stems of the plant cultivated in Cheju island, Korea, and examine their in vitro neuroprotective activities. Using a chromatographic fractionation method, ten chemical constituents were isolated from ethyl acetate extracts. By means of chemical and spectroscopic methods, those were identified as eight flavonoids such as kaempherol (a), quercetin (b), kaempferol 3-methyl ether (c), quercetin 3-methyl ether (d), narcissin (e), dihydrokaernpferol (f), dihydroquercetin (g) and erioclictyol (h), and two terpenoids such as 3-oxo-${\alpha}$-ionol-${\beta}$-d-glucopyranoside (i) and roseoside (j). Among the isolated compounds, comrounds c~e and h~j were those reported for the first titre from the plant. Compounds b, d and g showed DPPH free radical scavenging activities with IC$\sub$50/ values of 28, 19 and 31, ${\mu}$M respectively. Compounds d and g also inhibited iron-dependent lipid peroxidation with IC$\sub$50/ values of 2.4 and 3.5 ${\mu}$M. In a primary rat cortical neuronal cell culture system, compounds b, d and g inhibited xanthine/xanthine oxidase-induced (IC$\sub$50/ values of 18.2, 2.1 and 54.6 ${\mu}$M) and H$_2$O$_2$-induced (IC$\sub$50/ values of 13.6, 1.9 and 25.7 ${\mu}$M) cytotoxicities. In addition, compounds d and g inhibited NMDA-induced excitotoxicity by 21 and 33%, and only compound d inhibited growth factor withdrawal-induced apoptosis by 31% at a tested concentration of 3 ${\mu}$M. The results suggest that the antioxidant constituents with in vitroneuroprotective activities may serve as lead chemicals for the development of neuroprotective agent.

  • PDF

Effects of Plant Oils and Minerals for the Inhibition of Lipase Activity of Staphylococcus aureus Isolated from Fermented Pork Meat

  • Cho, Sang-Buem;Chang, Woo-Kyung;Kim, Yun-Jung;Moon, Hyung-In;Joo, Jong-Won;Choi, In-Soo;Seo, Kun-Ho;Kim, Soo-Ki
    • Food Science of Animal Resources
    • /
    • v.30 no.5
    • /
    • pp.764-772
    • /
    • 2010
  • Staphylococcus aureus lipase is regarded as a virulence factor. The response of lipase activity to various factors can provide important insights concerning the prevention of S. aureus during meat fermentation. This study was conducted to evaluate the main effects of nutrients used in culture media, and their combined effects on the inhibition of lipase activity and cell growth of pathogenic S. aureus SK1593 isolated from fermented pork meat. A Plackett-Burman design was used to evaluate the main effects of variables, including olive oil, soybean oil, grapeseed oil, sesame oil, $CuSO_4$, $MgCl_2$, $KNO_3$, $CaCl_2$, and KCl. Significant negative effects on lipase activity were detected with soybean oil, grapeseed oil, $KNO_3$, and $CaCl_2$. Additionally, these nutrients were further selected as variables for the investigation of their combined effect on lipase activity, via response surface methodology. In order to confirm the regression model, a situation that only inhibits lipase activity was simulated. The predicted lipase activity and cell growth of the simulated situation were 14.0 U/mL and $9.6\;{\log}_{10}$ (CFU/mL), respectively, and the estimated value of those in the same medium showed 15.14 U/mL and $9.4\;{\log}_{10}$(CFU/mL) respectively. The lipase activity of the simulated medium was inhibited approximately 5-fold as compared to the basal medium, but no significant differences in cell counts were noted to exist between the basal and simulated media. These results suggest that soybean oil, grapeseed oil, $KNO_3$, and $CaCl_2$ can be used to inhibit the growth of pathogenic S. aureus during the process of meat fermentation.

Microbial Communities of the Microbial Fuel Cell Using Swine Wastewater in the Enrichment Step with the Lapse of Time (가축분뇨를 이용한 미생물연료전지의 농화배양 단계에서 미생물 군집 변화)

  • Jang, Jae Kyung;Hong, Sun Hwa;Ryou, Youg Sun;Lee, Eun Young;Chang, In Seop;Kang, Young Koo;Kim, Jong Goo
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.35 no.12
    • /
    • pp.973-977
    • /
    • 2013
  • These studies were attempted to investigate the change of microbial community of anode of microbial fuel cell using swine wastewater in the enrichment step with the lapse of time. Microbial fuel cells enriched by a 1 : 1 mixture of anaerobic digestive juices of the sewage treatment plant and livestock wastewater. Enrichment culture step was divided into three stages to indentify the microorganisms. It was separated by each lag phase, exponential phase, and stationary phase. These steps were determined by the change of the current value. The current after enrichment was generated about $0.84{\pm}0.06mA$. We were cut out the different 17 bands in the DGGE fingerprint gel to do sequencing. The bands which the concentration was increasing or newly appearing with the lapse of time were included for this study. In the lag and exponential phase, Clostridium, Rhodocyclaceae, Bacteriodetes, and Uncultured bacterium etc. were detected. There were in the stationary phase Geobacter sp., Rhodocyclaceae, Candidatus, Nitrospira, Flavobactriaceae and uncultured bacterium etc. Geobactor among microorganisms detected in this study is known as the Electrochemically active microorganisms. It may include electrochemically active microorganisms to be considered as electrical activity microorganisms.

Fundamental Studies for the Breeding of Marine Algae 3. Effects of Plant Hormones on the Growth of Porphyra-fronds (해조의 육종을 위한 기초적 연구 3. 김 엽체에 미치는 식물호르몬의 생장효과)

  • KIM Joong-Rae;JO Young-Won;HAN Chang-Yeal;LEE Man-Sang
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.14 no.2
    • /
    • pp.79-85
    • /
    • 1981
  • As one of the fundamental studies for the breeding of marine algae, the effects of several plant hormones (IAA, Gibberellin, 2.4-D, NAA, Kinetin) on the growth of Porphyra-fronds, P. tenera Kjell. form tamatsuensis Miura, were investigated from January 21 to February 21 1981. The fronds used for the experiment were dissected out at $25mm^2$ size, and cultured in modified Provasoli's ESP medium supplemented with various concentrations of each plant growth regulators. The culture was kept under constant water temperature of $5^{\circ}C$ in 14 hrs. photoperiod and illuminated with 2,400 lux by fluorescent light. Based on the results of first experiment, the culture of fronds for the secondary experiment was carried out at $5^{\circ}C\;and\;10^{\circ}C$ in medium containing various levels of Kinetin from April 6 to 24, and compared the growth of two groups at each concentrations with each other, The results obtained are summarized as follows : (1) The best growth efficiencies were observed at 5.0mg/1 of each plant hormones except Gibberellin. Among them, the highest growth-rate was $312.5\%\;(345.3\%\;in\;frond\;size)$ in contrast with control at 5.0mg/1 of Kinetin, and was followed by $257.5\%\;(236.1\%)$ in 2.4-D,$166.7\%(147.6\%)$ in IAA and $141.7\%\;(167.7\%)$ in NAA, but that in Gibberellin was $247.9\%(241.9\%)$ at 10.0mg/l. (2) Especially, the fronds cultured at 5.0mg/1 of Kinetin were deep black-brown in colour, and had vivid, healthy chloroplasts in their all cells. On the contrary, the fronds cultured in other media were discoloured to light black-brown or green-drown, and almost all cells were vacuolated or shrunk gradually into death.(3) There was an obvious difference between the best growth-rates of the fronds cultured at 5.0mg/l of Kinetin at $5^{\circ}C$ and those at $10^{\circ}C$. The former was $366.7\%$, the latter $318.8\%$ but the difference was little at lower concentrations. (4) Many abnormal cells grown up to $25.0-27.5\mu$ in diameter were found among the marginal cells of fronds which showed the best growth in Kinetin, and the fronds wire $41.0-42.0\mu$ in thickness which was thicker by $10.0\mu$ or so than the others. (5) In two fronds at 1.0mg/1 of Kinetin cell-divisions were observed, which might developed into antheridium, but it was doubtful whether due to the efficiency of Kinetin.

  • PDF

Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
    • /
    • v.9 no.2
    • /
    • pp.156-165
    • /
    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

  • PDF

Effect of Antioxidant Flavonoids (Quercetin and Taxifolin) on In vitro Maturation of Porcine Oocytes

  • Kang, Jung-Taek;Moon, Joon Ho;Choi, Ji-Yei;Park, Sol Ji;Kim, Su Jin;Saadeldin, Islam M.;Lee, Byeong Chun
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.29 no.3
    • /
    • pp.352-358
    • /
    • 2016
  • Quercetin (QT) and taxifolin (TF) are structurally similar plant-derived flavonoids that have antioxidant properties and act as free radical scavengers. The objective of this study was to investigate effects of QT and TF on nuclear maturation of porcine oocytes. Effects of TF at 0, 1, 10, and $50{\mu}g/mL$ on oocyte nuclear maturation (polar body extrusion) were investigated. After incubation for 44 h, there were no significant differences between the treatment and control groups except in the $50{\mu}g/mL$ group which was significantly lower (59.2%, p<0.05) than the other groups (control: >80%). After parthenogenetic activation, further in vitro development of QT- or TF-treated vs control oocytes was investigated. A significantly higher proportion of QT-treated ($1{\mu}g/mL$) oocytes developed into blastocysts compared to controls (24.3% vs 16.8%, respectively); however, cleavage rate and blastocyst cell number were not affected. The TF-treated group was not significantly different from controls. Levels of reactive oxygen species (ROS) and intracellular glutathione (GSH) in oocytes and embryos in a culture medium supplemented with QT or TF were measured. Both treatment groups had significantly lower (p<0.05) levels of ROS than controls, however GSH levels were different only in QT-treated oocytes. We conclude that exogenous flavonoids such as QT and TF reduce ROS levels in oocytes. Although at high concentration ($50{\mu}g/mL$) both QT and TF appear to be toxic to oocytes.

Effect of Hypoxia on Carbohydrate Metabolism in Barley Seedlings (저산소 조건이 보리 유묘의 탄수화물대사에 미치는 영향)

  • Choi Heh Ran;Park Myoung Ryoul;Kim Jung Gon;Namkoong Seung Bak;Choi Kyeong-Gu;Yun Song Joong
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.50 no.3
    • /
    • pp.170-174
    • /
    • 2005
  • Barley plants growing in the wet paddy field easily encounter suboptimal oxygen concentration in the rhizosphere that causes molecular oxygen deficiency in root cells. The capacity of root cells to utilize energy sources is known to be positively related to resistance to hypoxia stress. This study was conducted to investigate effects of hypoxia on enzymes involved in the starch and sucrose metabolism. Barley seedlings at the third leaf stage were subjected to hypoxia (1 ppm dissolved oxygen) by purging the culture solution with nitrogen gas for up to seven days. The protein content was slightly decreased by hypoxia for 7 days. $\alpha-Amylase$ activities increased significantly in the root but not in the shoot after 3 to 7 days of hypoxia. $\beta-Amylase$ activities were not affected significantly in both tissues. Additionally, sucrose synthase activities were affected little in both tissues by 7 days of hypoxia. The results indicate that root cells activate break­down of polysaccharide reserves in response to an acute hypoxia to supply energy sources for fermentative glycolysis and cell wall fortification.

Durability of the Expanded Rice Hull as a Hydroponic Culture Medium (양액재배용 팽연화 왕겨의 적정 사용기간)

  • 임상현;김경희;안문섭;유근창
    • Journal of Bio-Environment Control
    • /
    • v.10 no.2
    • /
    • pp.106-110
    • /
    • 2001
  • In an effort to evaluate the economic value and durability of the expanded rice hull as substrates, changes in the physical and chemical properties of material and plant growth in that substrate were studied. Using and electron microscope, the structure of used and new expanded rice hull substrate was examined. Considerable decomposition was found in the substrate which had been used one to three times. Compactness and lowered porosity in the used substrates were probably caused by decomposition. The results of cation analysis showed the possible destruction of cell wall of rice hulls. Abundant $Ca^{2+}$ in the substrates used for two to three times also indicated the possibility of decomposition. In tomato yield comparison, 15.2% more yield of tomato fruit in a new substrates indicated the negative effects of decomposition of one-time used substrates. Yield decreased in the substrates used for three times. if perlite substrates is used for three years before renewal and the cost of the perlite renewal is counted. 65.3% saving in the cost will be realized with the use of an expanded rice hull substrate. Another positive effect of the expanded rice hull substrate is the decrease of environmental contamination.n.

  • PDF