• 제목/요약/키워드: Bacterial Inoculation

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Pseudomonas agarici와 P. toluaasii의 접종시기 및 접종농도가 팽이버섯의 균사생장과 자실체형성에 미치는 영향 (Effect of Inoculation Time and Population Density of Pseudomonas agarici and P. tolaasii on the Mycelial Growth and the Fruit body Formation in Flammulina velutipes)

  • 이현욱;문병주;이흥수;차흥오;조동진;신원교
    • 한국균학회지
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    • 제26권1호통권84호
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    • pp.60-68
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    • 1998
  • 버섯류에 세균병을 유발시키는 Pseudomonas agarici와 P. tolaasii가 팽이버섯균의 균사생장 및 자실체형성에 미치는 영향을 알아보기 위해 접종시기와 접종농도를 달리하여 기내에서 조사하였다. 팽이버섯균과 병원세균을 동시에 접종하거나 팽이버섯균접종 5일후에 병원세균을 접종했을 때 병원세균의 접종농도가 높아짐에 따라 균사생장과 자실체 형성이 현저하게 억제되는 경향이었으며, 특히 $10^2{\sim}10^6\;cfu/g$ media의 접종농도에서 균사생장이 지연되면서 엷음 또는 흐림증상을 보였고 $10^8\;cfu/g$ media농도에서는 특정분위에서부터 균사가 더이상 진전되지 못하는 멈춤증상을 나타냈다. 그리고 P. agarici $10^2\;cfu/g$ media 이상의 농도와 P. tolaasii $10^4\;cfu/g$ media 이상의 농도에서는 자실체의 수확을 전혀 기대할 수 없었다. 두 병원세균 사이의 팽이버섯균에 대한 균사생장억제력은 P. agarici에 비해 P. tolaasii가 다소 높았으며, 자실체형성억제력은 P. tolaasii 비해 P. agarici가 다소 높은 결과를 보였다. 그러나 팽이버섯균접종 10일후 병원세균을 접종한 경우 두 병원세균 모두가 균사생장과 자실체생육에 거의 영향을 미치지 않는 것으로 나타났다.

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Expression and Promoter Analyses of Pepper CaCDPK4 (Capsicum annuum calcium dependent protein kinase 4) during Plant Defense Response to Incompatible Pathogen

  • Chung, Eun-Sook;Oh, Sang-Keun;Park, Jeong-Mee;Choi, Do-Il
    • The Plant Pathology Journal
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    • 제23권2호
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    • pp.76-89
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    • 2007
  • CaCDPK4, a full-length cDNA clone encoding Capsicum annuum calcium-dependent protein kinase 4, was isolated from chili pepper (Capsicum annuum L.). Deduced amino acid sequence of CaCDPK4 shares the highest homology with tobacco NpCDPK8 and chickpea CaCDPK2 with 79% identity. Genomic blot analyses revealed that CaCDPK4 is present as a single copy in pepper genome, but it belongs to a multigene family. CaCDPK4 was highly induced when pepper plants were inoculated with an incompatible bacterial pathogen. Induced levels of CaCDPK4 transcripts were also detected in pepper leaves by the treatment of ethephon, an ethylene-inducing agent, and high-salt stress condition. The bacterial-expressed GST-CaCDPK4 protein showed to retain the autophosphorylation activity in vitro. GUS expression driven by CaCDPK4 promoter was examined in transgenic Arabidopsis containing transcriptional fusion of CaCDPK4 promoter. GUS expression under CaCDPK4 promoter was strong in the root and veins of the seedlings. GW (-1965) and D3 (-1377) promoters conferred on GUS expression in response to inoculation of an incompatible bacterial pathogen, but D4-GUS (-913) and DS-GUS (-833) did not. Taken together, our results suggest that CaCDPK4 can be implicated on signal transduction pathway of defense response against an incompatible bacterial pathogen in pepper.

Enhancement of Drought-Stress Tolerance of Brassica oleracea var. italica L. by Newly Isolated Variovorax sp. YNA59

  • Kim, Yu-Na;Khan, Muhammad Aaqil;Kang, Sang-Mo;Hamayun, Muhammad;Lee, In-Jung
    • Journal of Microbiology and Biotechnology
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    • 제30권10호
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    • pp.1500-1509
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    • 2020
  • Drought is a major abiotic factor and has drastically reduced crop yield globally, thus damaging the agricultural industry. Drought stress decreases crop productivity by negatively affecting crop morphological, physiological, and biochemical factors. The use of drought tolerant bacteria improves agricultural productivity by counteracting the negative effects of drought stress on crops. In this study, we isolated bacteria from the rhizosphere of broccoli field located in Daehaw-myeon, Republic of Korea. Sixty bacterial isolates were screened for their growth-promoting capacity, in vitro abscisic acid (ABA), and sugar production activities. Among these, bacterial isolates YNA59 was selected based on their plant growth-promoting bacteria traits, ABA, and sugar production activities. Isolate YNA59 highly tolerated oxidative stress, including hydrogen peroxide (H2O2) and produces superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities in the culture broth. YNA59 treatment on broccoli significantly enhanced plant growth attributes, chlorophyll content, and moisture content under drought stress conditions. Under drought stress, the endogenous levels of ABA, jasmonic acid (JA), and salicylic acid (SA) increased; however, inoculation of YNA59 markedly reduced ABA (877 ± 22 ng/g) and JA (169.36 ± 20.74 ng/g) content, while it enhanced SA levels (176.55 ± 9.58 ng/g). Antioxidant analysis showed that the bacterial isolate YNA59 inoculated into broccoli plants contained significantly higher levels of SOD, CAT, and APX, with a decrease in GPX levels. The bacterial isolate YNA59 was therefore identified as Variovorax sp. YNA59. Our current findings suggest that newly isolated drought tolerant rhizospheric Variovorax sp. YNA59 is a useful stress-evading rhizobacterium that improved drought-stress tolerance of broccoli and could be used as a bio-fertilizer under drought conditions.

Isolation, Characterization, and Use for Plant Growth Promotion Under Salt Stress, of ACC Deaminase-Producing Halotolerant Bacteria Derived from Coastal Soil

  • Siddikee, M.A.;Chauhan, P.S.;Anandham, R.;Han, Gwang-Hyun;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • 제20권11호
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    • pp.1577-1584
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    • 2010
  • In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

Exploring the Potentiality of Novel Rhizospheric Bacterial Strains against the Rice Blast Fungus Magnaporthe oryzae

  • Amruta, Narayanappa;Kumar, M.K. Prasanna;Puneeth, M.E.;Sarika, Gowdiperu;Kandikattu, Hemanth Kumar;Vishwanath, K.;Narayanaswamy, Sonnappa
    • The Plant Pathology Journal
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    • 제34권2호
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    • pp.126-138
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    • 2018
  • Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

Xanthomonas axonopodis pv. eucalyptorum pv. nov. Causing Bacterial Leaf Blight on Eucalypt in Brazil

  • Ferraz, Helvio Gledson Maciel;Badel, Jorge Luis;da Silva Guimaraes, Lucio Mauro;Reis, Bruna Paolinelli;Totola, Marcos Rogerio;Goncalves, Rivadalve Coelho;Alfenas, Acelino Couto
    • The Plant Pathology Journal
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    • 제34권4호
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    • pp.269-285
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    • 2018
  • Bacterial leaf blight is a major disease of eucalypt, especially under nursery conditions. Different bacterial species have been associated with the disease in several countries, and despite its importance worldwide, it is not clear to date whether similar disease symptoms are caused by the same or by different etiological agents. In this study, 43 bacterial strains were isolated from blighted eucalypt leaves collected in different geographic areas of Brazil and inoculated onto a susceptible eucalypt clone. Polyphasic taxonomy, including morphological, physiological, biochemical, molecular, and pathogenicity tests showed that only certain strains of Xanthomonas axonopodis caused symptoms of the disease. Strains varied in their aggressiveness, but no correlation with geographic origin was observed. MLSA-based phylogenetic analysis using concatenated dnaK, fyuA, gyrB and rpoD gene sequences allocated the strains in a well-defined clade, corresponding to Rademarker's group RG 9.6. Inoculation of nineteen plant species belonging to seven botanical families with representative strain LPF 602 showed it to be pathogenic only on Eucalyptus spp, and Corymbia spp. Based on distinct biochemical and pathogenic characteristics that differentiate the eucalypt strains from other pathovars of the X. axonopodis species, here we propose their allocation into the new pathovar X. axonopodis pv. eucalyptorum pv. nov.

Improved dentin disinfection by combining different-geometry rotary nickel-titanium files in preparing root canals

  • Bedier, Marwa M.;Hashem, Ahmed Abdel Rahman;Hassan, Yosra M.
    • Restorative Dentistry and Endodontics
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    • 제43권4호
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    • pp.46.1-46.10
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    • 2018
  • Objectives: This study was to evaluate the antibacterial effect of different instrumentation and irrigation techniques using confocal laser scanning microscopy (CLSM) after root canal inoculation with Enterococcus faecalis (E. faecalis). Materials and Methods: Mesiobuccal and mesiolingual canals of extracted mandibular molars were apically enlarged up to a size 25 hand K-file, then autoclaved and inoculated with E. faecalis. The samples were randomly divided into 4 main groups according to the system of instrumentation and irrigation: an XP-endo Shaper (XPS) combined with conventional irrigation (XPS/C) or an XP-endo Finisher (XPF) (XPS/XPF), and iRaCe combined with conventional irrigation (iRaCe/C) or combined with an XPF (iRaCe/XPF). A middle-third sample was taken from each group, and then the bacterial reduction was evaluated using CLSM at a depth of $50{\mu}m$ inside the dentinal tubules. The ratio of red fluorescence (dead cells) to green-and-red fluorescence (live and dead cells) represented the percentage of bacterial reduction. The data were then statistically analyzed using the Kruskal-Wallis test for comparisons across the groups and the Dunn test was used for pairwise comparisons. Results: The instrumentation and irrigation techniques had a significant effect on bacterial reduction (p < 0.05). The iRaCe/XPF group showed the strongest effect, followed by the XPS/XPF and XPS/C group, while the iRaCe/C group had the weakest effect. Conclusions: Combining iRaCe with XPF improved its bacterial reduction effect, while combining XPS with XPF did not yield a significant improvement in its ability to reduce bacteria at a depth of $50{\mu}m$ in the dentinal tubules.

Identification and Characterization of Xanthomonas arboricola pv. juglandis Causing Bacterial Blight of Walnuts in Korea

  • Kim, Hyun Sup;Cheon, Wonsu;Lee, Younmi;Kwon, Hyeok-Tae;Seo, Sang-Tae;Balaraju, Kotnala;Jeon, Yongho
    • The Plant Pathology Journal
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    • 제37권2호
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    • pp.137-151
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    • 2021
  • The present study describes the bacterial blight of walnut, caused by Xanthomonas arboricola pv. juglandis (Xaj) in the northern Gyeongbuk province, Korea. Disease symptoms that appear very similar to anthracnose symptoms were observed in walnut trees in June 2016. Pathogens were isolated from disease infected leaves, fruits, shoots, bud, flower bud of walnut, and cultured onto yeast dextrose carbonate agar plates. Isolated bacteria with bacterial blight symptoms were characterized for their nutrient utilization profiles using Biolog GN2 and Vitek 2. In addition, isolates were subjected to physiological, biochemical, and morphological characterizations. Furthermore, isolates were identified using 16S rDNA sequence analysis, and multi-locus sequence analysis using atpD, dnaK, efp, and rpoD. To confirm pathogenicity, leaves, fruits, and stems of 3-year-old walnut plants were inoculated with bacterial pathogen suspensions as a foliar spray. One week after inoculation, the gray spots on leaves and yellow halos around the spots were developed. Fruits and stems showed browning symptoms. The pathogen Xaj was re-isolated from all symptomatic tissues to fulfill Koch's postulates, while symptoms were not appeared on control plants. On the other hand, the symptoms were very similar to the symptoms of anthracnose caused by Colletotrichum gloeosporioides. When walnut plants were inoculated with combined pathogens of Xaj and C. gloeosporioides, disease symptoms were greater in comparison with when inoculated alone. Xaj population size was more in the month of April than March due to their dormancy in March, and sensitive to antibiotics such as oxytetracycline and streptomycin, while resistant to copper sulfate.

Inhibitory Effects of Pepper Mild Mottle Virus Infection by Supernatants of Five Bacterial Cultures in Capsicum annuum L.

  • Venkata Subba Reddy, Gangireddygari;In-Sook, Cho;Sena, Choi;Ju-Yeon, Yoon
    • The Plant Pathology Journal
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    • 제38권6호
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    • pp.646-655
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    • 2022
  • Pepper mild mottle virus (PMMoV), one of the most prevalent viruses in chili pepper (Capsicum annuum L.) is a non-enveloped, rod-shaped, single-stranded positive-sense RNA virus classified in the genus Tobamovirus. The supernatants of five bacterial cultures (Pseudomonas putida [PP], Bacillus licheniformis [BLI], P. fluorescens [PF], Serratia marcescens [SER], and B. amyloliquifaciens [BA]) were analyzed to find novel antiviral agents to PMMoV in chili pepper. Foliar spraying with supernatants (1:1, v/v) obtained from Luria-Bertani broth cultures of PP, BLI, PF, SER, and BA inhibited PMMoV infection of chili pepper if applied before the PMMoV inoculation. Double-antibody sandwich enzyme-linked immunosorbent assay showed that treatments of five supernatants resulted in 51-66% reductions in PMMoV accumulation in the treated chili pepper. To identify key compounds in supernatants of PP, BLI, PF, SER, and BA, the supernatants were subjected to gas chromatography-mass spectrometry. The 24 different types of compounds were identified from the supernatants of PP, BLI, PF, SER, and BA. The compounds vary from supernatants of one bacterial culture to another which includes simple compounds-alkanes, ketones, alcohols, and an aromatic ring containing compounds. The compounds triggered the inhibitory effect on PMMoV propagation in chili pepper plants. In conclusion, the cultures could be used to further conduct tissue culture and field trial experiments as potential bio-control agents.

해양퇴적토내 다환방향족탄화수소 생분해 증진 조건 연구 (Condition of ex situ Bioremediation of Polycyclic Aromatic Hydrocarbons in Marine Sediments)

  • 정홍배;;이희순;권개경;김상진
    • 한국해양환경ㆍ에너지학회지
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    • 제8권4호
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    • pp.179-185
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    • 2005
  • 다환방향족 탄화수소는 주요한 독성환경오염물질풍의 하나이며 해양퇴적물에 축적이 된다. 다환방향족 탄화수소가 가지고 있는 위해성으로 인해 해양퇴적물내의 다환방향족 탄화수소 제거는 많은 관심사가 되어오고 있다. 본 연구에서는 다환방향족 탄화수소로 오염된 해양퇴적물의 생물정화를 위해 bioslurry reactor를 사용하기 위한 전단계로 ex situ bioremediation을 위한 최적 조건을 찾기 위해 멸균 유무의 퇴적물에서 균주접종과 혼합 cyclodextrin (M-CD)공급의 효과를 알아보았다. 비멸균상태의 퇴적물에서 분해균주의 접종여부에 상관없이 M-CD가 첨가된 실험구의 전자전달계활성이 증가되었고 멸균상태의 퇴적물에서는 분해균주가 접종되고 M-CD가 첨가된 실험구의 전자전달계활성이 증가되었다. 멸균상태의 퇴적물에서 M-CD와 함께 단일 균주가 접종되었을 때 퇴적물내의 다환방향족 탄화수소의 분해율이 9-20%로 나타났으며 동일한 조건에서 혼합균주가 접종되었을 때는 24-37%로 나타났다. 비멸균상태의 퇴적물에서 동일한 조건으로 실험하였을 때는 유의한 분해가 일어나지 않았으며 군집분석 결과 접종 균주도 감소하였다. 또한 M-CD 첨가 없이는 2일후 접종균주를 찾아볼 수 없었다. 결론적으로 퇴적물내의 다환방향족 탄화수소 제거를 위해 퇴적물내의 토착미생물과 포식자가 제거된 상태에서 적절한 탄소원과 함께 균주를 접종하는 것이 효과적인 것으로 나타났다.

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