• Title/Summary/Keyword: Bacterial Inoculation

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Evaluation of Bioremediation Effectiveness by Resolving Rate-Limiting Parameters in Diesel-Contaminated Soil

  • Joo, Choon-Sung;Oh, Young-Sook;Chung, Wook-Jin
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.607-613
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    • 2001
  • The biodegradation rates of diesel oil by a selected diesel-degrading bacterium, Pseudomonas stutzeri strain Y2G1, and microbial consortia composed of combinations of 5 selected diesel-degrading bacterial were determined in liquid and soil systems. The diesel degradation rate by strain Y2G1 linearly increased $(R^2=0.98)$ as the diesel concentration increased up to 12%, and a degradation rate as high as 5.64 g/l/day was obtained. The diesel degradation by strain Y2G1 was significantly affected by several environmental factors, and the optimal conditions for pH, temperature, and moisture content were at pH8, $25^{\circ}C$, and 10%, respectively. In the batch soil microcosm tests, inoculation, especially in the form of a consortium, and the addition of nutrients both significantly enhanced the diesel degradation by a factor of 1.5 and 4, respectively. Aeration of the soil columns effectively accelerated the diesel degradation, and the initial degradation rate was obviously stimulated with the addition of inorganic nutrients. Based on these results, it was concluded that the major rate-limiting factors in the tested diesel-contaminated soil were the presence of inorganic nutrients, oxygen, and diesel-degrading microorganisms. To resolve these limiting parameters, bioremediation strategies were specifically designed for the tested soil, and the successful mitigation of the limiting parameters resulted in an enhancement of the bioremediation efficiency by a factor of 11.

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Salt Tolerance in Transgenic Pea (Pisum sativum L.) Plants by P5CS Gene Transfer

  • Najafi F.;Rastgar-jazii F.;Khavari-Nejad R. A.;Sticklen M.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.233-240
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    • 2005
  • Slices of embryonic axis of mature pea (Pisum sativum L. cv. Green Arrow) seeds were used as explant. Transformation of explants was done via Agrobacterium tumefaciens bearing vector pBI-P5CS construct. The best results for inoculation of explants were obtained when they were immersed for 90 s at a concentration of $6{\times}10^8$ cell $ml^(-1)$ of bacterial suspension. Transformed pea plants were selected on $50\;mg\;l^(-1)$ kanamycin and successful transformants were confirmed by PCR and blotting. Transgenic plants were further analyzed with RT-PCR to confirm the expression of P5CS. Transgenic plants and non-transgenic plants were treated with different concentrations of NaCl 0 (control), 100, 150 and 200 mM in culture medium. Measurement of proline content indicated that transgenic plants produced more amino acid proline in response to salt in comparison with non-transgenic plants. Photosynthetic efficiency in transgenic plants under salt-stress was more than that of non-transgenic plants.

Enterobacter cloacae, an Emerging Plant-Pathogenic Bacterium Affecting Chili Pepper Seedlings

  • Garcia-Gonzalez, Tanahiri;Saenz-Hidalgo, Hilda Karina;Silva-Rojas, Hilda Victoria;Morales-Nieto, Carlos;Vancheva, Taca;Koebnik, Ralf;Avila-Quezada, Graciela Dolores
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.1-10
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    • 2018
  • A previously unreported bacterial disease on chili pepper (Capsicum annuum L.) seedlings affecting as many as 4% of seedlings was observed in greenhouses in Chihuahua, Mexico (Delicias and Meoqui counties). Initial lesions appeared as irregular small spots on leaves and brown necrosis at margins tips were observed. Later, the spots became necrotic with a chlorotic halo. Advanced disease was associated with defoliation. A Gram negative, rod-shaped bacterium was isolated from diseased chili pepper seedlings. Three inoculation methods revealed that isolated strains produce foliage symptoms, similar to those observed in naturally infected seedlings. Pathogenic strains that caused symptoms in inoculated seedlings were re-isolated and identified to fulfill koch's postulate. Polyphasic approaches for identification including biochemical assays (API 20E and 50CH), carbon source utilization profiling (Biolog) and 16S rDNA, hsp60 and rpoB sequence analysis were done. Enterobacter cloacae was identified as the causal agent of this outbreak on chili pepper seedlings.

Occurrence of crown gall of chrysanthemum caused by Agrobacterium tumefaciens.

  • Lee, Young-Kee;Lee, Jong-Hyoung;Kim, Jin-Young;Cho, Weon-Dae;Cha, Jae-Soon
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.126-126
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    • 2003
  • Incidence of crown gall on lower stem of chrysanthemum, Chrysanthemum morifolium Ramat., was first observed at Hwasung, Gyeonggi, Korea in 2001, Tumors on the stem were 1.5-2 cm in size and semi-round with rough surface texture of dark brown color. Four strains of bacteria isolated from the tumor tissues were characterized. Their colonies were convex, glistening, circular with an entire edge, and white to tannish-cream in color on PDA plus CaCO$_3$. They were gram negative, oxidase positive, and growing on DIM agar. The bacterial isolates inducing gall formation in chrysanthemum were identified as Agrobacterium tumefaciens based on biochemical and physiological characteristics, fatty acid profile using Sherlock Microbial Identification System, and substrate utilization patterns using Biolog Identification System. Young chrysanthemum plants inoculated with the bacteria developed typical galls within two to three weeks. Seedlings of tomato and slices of carrot roots also produced typical galls two to three weeks after inoculation. This is the first report on crown gall of chrysanthemum in Korea.

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Hypersensitive and Apoptotic Responses of Pepper Fruit Against Xnthomonas axonopodis pv. glycines Infection

  • Chang, Sung-Pae;Kim, Young-Ho
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.72.1-72
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    • 2003
  • Generally, plants defend themselves against pathogens by structural and biochemical reactions. Defense structures act as physical barriers and inhibit the pathogen from gaining entrance and spreading through the plant. Xanthomonas axonopodis pv glycines, the causal pathogen of bacterial pustule of soybean, causes hypersensitive response (HR). When pepper fruits were inoculated with X. axonopodis pv. glycines, in situ, time-series defense-related structural changes occurred in the inoculated sites. Early responses were programmed cell death (PCD), characterized by condensation and vacuolization of the cytoplasm, condensation of nuclear materials, and fragmentation of the nuclear DNA, which were observed by transmission electron microscopy. Nuclear fragmentation was proven by TUNEL method under confocal laser scanning microscopy and DNA laddering through eletrophoresis. At later stages, plant responses were cell elongation and cell division, forming a periderm-like boundary layer that demarcated healthy tissues from the inoculation sites. Using several stains such as toluidine blue, sudan IV, annexin V, and phloroglucinol-HCl, defense-related materials and structural changes were also examined.

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Activation of Defense Responses in Chinese Cabbage by a Nonhost Pathogen, Pseudomonas syringae pv. tomato

  • Park, Yong-Soon;Jeon, Myeong-Hoon;Lee, Sung-Hee;Moon, Jee-Sook;Cha, Jae-Soon;Kim, Hak-Yong;Cho, Tae-Ju
    • BMB Reports
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    • v.38 no.6
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    • pp.748-754
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    • 2005
  • Pseudomonas syringae pv. tomato (Pst) causes a bacterial speck disease in tomato and Arabidopsis. In Chinese cabbage, in which host-pathogen interactions are not well understood, Pst does not cause disease but rather elicits a hypersensitive response. Pst induces localized cell death and $H_2O_2$ accumulation, a typical hypersensitive response, in infiltrated cabbage leaves. Pre-inoculation with Pst was found to induce resistance to Erwinia carotovora subsp. carotovora, a pathogen that causes soft rot disease in Chinese cabbage. An examination of the expression profiles of 12 previously identified Pst-inducible genes revealed that the majority of these genes were activated by salicylic acid or BTH; however, expressions of the genes encoding PR4 and a class IV chitinase were induced by ethephon, an ethylene-releasing compound, but not by salicylic acid, BTH, or methyl jasmonate. This implies that Pst activates both salicylate-dependent and salicylate-independent defense responses in Chinese cabbage.

Inhibitory Effects of Acinetobacter sp. KTB3 on Infection of Tobacco mosaic virus in Tobacco Plants

  • Kim, Young-Sook;Hwang, Eui-ll;O, Jeong-Hun;Kim, Kab-Sig;Ryu, Myong-Hyun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.293-296
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    • 2004
  • During the screening of antiviral substances having inhibitory effects on Tobacco mosaic virus (TMV) infection on tobacco plants, we found a bacterial isolate KTB3, and identified it as Acinetobacter sp. which strongly inhibited the infection of TMV When the culture filtrate from KTB3 was applied on the upper surface of the Xanthi-nc tobacco leaves at the same time, or 24 hours before TMV inoculation, almost complete inhibition was achieved. Likewise, 86% inhibition was achieved, when the culture filtrate was applied on the underside of the leaves. In field trials, transmission of TMV from diseased seedlings to healthy ones during transplanting work was reduced by 92%, when the culture filtrate was sprayed onto the tobacco seedlings, cv. NC82, 24 hours before transplanting. No toxic effect was observed on the tobacco plants. Antiviral substance from the culture filtrate was purified by ethanol precipitation, dialysis, DEAE-cellulose, and Sephadex G75 gel column chromatography. The partially purified active material which showed positive color reaction to sugar and protein inhibited TMV infection by 60% at 1 ${\mu}$g/ml.

Monitoring of Microorganisms Added into Oil-Contaminated Microenvironments by Terminal-Restriction Fragment Length Polymorphism Analysis

  • JUNG SEONG-YOUNG;LEE JUNG-HYUN;CHAI YOUNG-GYU;KIM SANG-JIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1170-1177
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    • 2005
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

Identification of Novel Bioactive Hexapeptides Against Phytopathogenic Bacteria Through Rapid Screening of a Synthetic Combinatorial Library

  • Choi, Jae-Hyuk;Moon, Eun-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.792-802
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    • 2009
  • Antimicrobial peptides (AMPs) are considered to be a promising alternative to conventional antibiotics for future generations. We identified four novel hexapeptides with antimicrobial activity: KCM11 (TWWRWW-$NH_2$), KCM12 (KWRWlW-$NH_2$), KCM21 (KWWWRW-$NH_2$), and KRS22 (WRWFIH-$NH_2$), through positional scanning of a synthetic peptide combinatorial library (PS-SCL). The ability of these peptides to inhibit the growth of a variety of bacteria and unicellular fungi was evaluated. KCM11 and KRS22 preferentially inhibited the normal growth of fungal strains, whereas KCM12 and KCM21 were more active against bacterial strains. Bactericidal activity was addressed in a clear zone assay against phytopathogenic bacteria, including Pectobacterium spp., Xanthomonas spp., Pseudomonas spp., etc. KCM21 showed the highest activity and was effective against a wide range of target organisms. Application of KCM21 with inoculation of Pectobacterium carotovorum subsp. carotovorum on detached cabbage leaves resulted in an immune phenotype or a significant reduction in symptom development, depending on the peptide concentration. Cytotoxicity of the four hexapeptides was evaluated in mouse and human epithelial cell lines using an MTT test. The results revealed a lack of cytotoxic effects.

Rumen Manipulation to Improve Animal Productivity

  • Santra, A.;Karim, S.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.748-763
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    • 2003
  • Anaerobic rumen microorganisms mainly bacteria, protozoa and fungi degrade ligno-cellulosic feeds consumed by the ruminants. The ruminants in developing countries are predominantly maintained on low grade roughage and grazing on degraded range land resulting in their poor nutrient utilization and productivity. Hence, manipulation of rumen fermentation was tried during last two decades to optimize ruminal fermentation for improving nutrient utilization and productivity of the animals. Modification of rumen microbial composition and their activity was attempted by using chemical additives those selectively effect rumen microbes, introduction of naturally occurring or genetically modified foreign microbes into the rumen and genetically manipulation of existing microbes in the rumen ecosystem. Accordingly, rumen protozoa were eliminated by defaunation for reducing ruminal methane production and increasing protein outflow in the intestine, resulting in improve growth and feed conversion efficiency of the animals. Further, Interspecies trans-inoculation of rumen microbes was also successfully used for annulment of dietary toxic factor. Additionally, probiotics of bacterial and yeast origin have been used in animal feeding to stabilize rumen fermentation, reduced incidence of diarrhoea and thus improving growth and feed conversion efficiency of young stalk. It is envisaged that genetic manipulation of rumen microorganisms has enormous research potential in developing countries. In view of feed resource availability more emphasis has to be given for manipulating rumen fermentation to increase cellulolytic activity for efficient utilization of low grade roughage.