• Title/Summary/Keyword: strains inoculation

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Cytotoxic, Antioxidative, and ACE Inhibiting Activities of Dolsan Leaf Mustard Juice (DLMJ) Treated with Lactic Acid Bacteria

  • Yoo Eun-Jeong;Lim Hyun-Soo;Park Kyung-Ok;Choi Myeong-Rak
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.60-66
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    • 2005
  • This study was performed to know whether there is any change of physiological activity in DLMJ which is inoculated by lactic acid bacteria. Lactic acid bacteria were isolated from Dolsan leaf mustard Kimchi (DLMK) at $20^{\circ}C$. In the optimum ripening period, the population of Leuconostoc and Lactobacilli in the DLMK were found to be high. The Leuconostoc, Lactobacilli and Lactococci strains were identified as Leuconostoc mesenteroides, Leuconostoc gelidum, Weissella confusa, Lactobacillus plantarum, Lactobacillus raffinolactis, Lactococcus lactis and Weissella confusa using the Biolog system. The most predominant strain which was isolated from DLMK was Weissella confusa. As the results of the phylogenetic analysis using 16s rDNA sequence, the Weissella confusa turned out to be Weissella kimchii, with 99.0% similarity. To investigated the change of physiological activity in DLMJ by lactic acid bacteria, 7 predominant strains inoculated to DLMJ (Dolsan Leaf Mustard Juice). The cytotoxicity was found to be under $19.55\%$ all cases. Also, the antioxidative activity of the DLMJ treated with lactic acid bacteria was very low, which might have been due to the reduced antioxidative phytochemicals during the preparation of the sterile sample. The ACE inhibiting activity of DLMJ by inoculation with Weissella kimchii was shown to be the highest ($94.0\%$). This could be that the degradation of sulfur containing materials in DLMJ by Weissella kimchii gave rise to ACE inhibiting activity.

Role of Two Sets of RND-Type Multidrug Efflux Pump Transporter Genes, mexAB-oprM and mexEF-oprN, in Virulence of Pseudomonas syringae pv. tabaci 6605

  • Ichinose, Yuki;Nishimura, Takafumi;Harada, Minori;Kashiwagi, Ryota;Yamamoto, Mikihiro;Noutoshi, Yoshiteru;Toyoda, Kazuhiro;Taguchi, Fumiko;Takemoto, Daigo;Matsui, Hidenori
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.148-156
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    • 2020
  • Pseudomonas syringae pv. tabaci 6605 has two multidrug resistance (MDR) efflux pump transporters, MexAB-OprM and MexEF-OprN. To understand the role of these MDR efflux pumps in virulence, we generated deletion mutants, ΔmexB, ΔmexF, and ΔmexBΔmexF, and investigated their sensitivity to plant-derived antimicrobial compounds, antibiotics, and virulence. Growth inhibition assays with KB soft agar plate showed that growth of the wild-type (WT) was inhibited by 5 μl of 1 M catechol and 1 M coumarin but not by other plant-derived potential antimicrobial compounds tested including phytoalexins. The sensitivity to these compounds tended to increase in ΔmexB and ΔmexBΔmexF mutants. The ΔmexBΔmexF mutant was also sensitive to 2 M acetovanillone. The mexAB-oprM was constitutively expressed, and activated in the ΔmexF and ΔmexBΔmexF mutant strains. The swarming and swimming motilities were impaired in ΔmexF and ΔmexBΔmexF mutants. The flood inoculation test indicated that bacterial populations in all mutant strains were significantly lower than that of WT, although all mutants and WT caused similar disease symptoms. These results indicate that MexAB-OprM extrudes plant-derived catechol, acetovanillone, or coumarin, and contributes to bacterial virulence. Furthermore, MexAB-OprM and MexEF-OprN complemented each other's functions to some extent.

Biological and Physicochemical Properties of Canine Parvovirus lated from the Dogs with Diarrhea (설사증 나환견으로 부터 분리한 Canine Parvovirus의 성상에 관한 연구)

  • 최해연;정운선;전무형;박성국;민원기
    • Korean Journal of Veterinary Service
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    • v.13 no.2
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    • pp.162-183
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    • 1990
  • From 1988 to 1989, 8 strains of canine parvovirus-2 (CPV-2) were isolated from the fecal specimens from the dogs that were clinically diagnosed as canine parvoviral enteritis in the veterinary hospitals located in the regions of Taejon and Chungbuk province. Studios on biological and physicochemical properties for the isolates were carried out. The results obtained by experiments are summarized as follows. 1. Among 62 fecal samples collected from the dogs with enteric diseases, 24 (38.7%) showed the haemagglutinating activity against porcine erythrocyte, ranging from 16 to 16, 384 of HA titers. 2. When 8 fecal specimens with high HA titer over 1, 000 were inocultated into CRFX cells, intranuclear inclusion bodies were obseverd in all of eight specimens, of w)lick three specimens showed cytoplasmic inclusions concurrently with the intranuclear inclusion bodies. 3. In study on species-specificity of haemagglutinating activity of the isolates, TJ-89-1 and TJ-89-2, it was found that the isolates revealed the highest haemagglutinating activity with porcine erythrocytes, showing the relatively lower haemagglutination titers with the erythrocytes from cat and rabbit. None of erythrocytes from other animals reacted with the isolates. 4. By the cross-haemagglutination inhibition test of the Isolates with reference viruses and sera, the Isolates were evidently identified as the strains of canine parvovirus-2. 5. In Physicochemical property test, it was evident that the isolates were stable in, lipid solvent, pH and heat treatment at $56^{\circ}C$ for 30 min. and contain DNA genome. 6. When seven puppies were inoculated intraorally with the isolate at HA titer of 8, 192, all of the puppies showed the symptoms of anorexia, vomiting, diarrhea and died at the 5th to 10th days post inoculation(pi). The fecal samples from all of the puppies revealed significantly high HA titers afterward the 5th days pi. Body temperature and the number of total leucocytes were slightly increased at the early stage of infection. but extremely decreased at the stage of collapse. HI titers of the sera started to increase at the 2nd to 3rd days pi reaching 512 to 1, 024 at the 4th to 5th day pi.

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Assessment of Root-Associated Paenibacillus polymyxa Groups on Growth Promotion and Induced Systemic Resistance in Pepper

  • Phi, Quyet-Tien;Park, Yu-Mi;Seul, Keyung-Jo;Ryu, Choong-Min;Park, Seung-Hwan;Kim, Jong-Guk;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1605-1613
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    • 2010
  • Twenty-nine P. polymyxa strains isolated from rhizospheres of various crops were clustered into five genotypic groups on the basis of BOX-PCR analysis. The characteristics of several plant growth-promoting factors among the isolates revealed the distinct attributes in each allocated group. Under gnotobiotic conditions, inoculation of pepper roots with P. polymyxa isolates significantly increased the biomass in 17 of total 29 treated plants with untreated plants. Experiments on induced systemic resistance (ISR) against bacterial spot pathogen Xanthomonas axonopodis pv. vesicatoria in pepper by P. polymyxa strains were conducted and only one isolate (KNUC265) was selected. Further studies into ISR mediation by the KNUC265 strain against the soft-rot pathogen Erwinia carotovora subsp. carotovora in tobacco demonstrated that the tobacco seedlings exposed to either bacterial volatiles or diffusible metabolites exhibited a reduction in disease severity. In conclusion, ISR and plant growth promotion triggered by P. polymyxa isolates were systemically investigated on pepper for the first time. The P. polymyxa KNUC265 strain, which elicited both ISR and plant growth promotion, could be potentially used in improving the yield of pepper and possibly of other crops.

In Situ Detection and Differential Counts of Bifidobacterium spp. Using Bromocresol Green, a pH-dependent Indicator

  • Kim, Ki-Hwan;Shin, Won-Cheol;Park, Young-Seo;Yoon, Sung-Sik
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.99-103
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    • 2007
  • The purpose of this study was to develop a simple detection method, possibly at the species-level, that allows for large-scale screening of bifidobacteria. Human fecal samples were plated on MRS-raffinose agar containing cysteine and neomycin sulfate, serving as selective pressure for bifidobacteria, and 0.003%(w/v) bromocresol green. All of the test strains grew well on this medium at $37{\pm}1^{\circ}C$, forming white colonies surrounded by yellow halos, which presented a sharp contrast against the green background. In this disc assay, the required incubation time to develop a yellowish zone varied with the species of Bifidobacterium that was tested, allowing for differential counts and easy identification at the species-level: 10-14 hr for B. bifidum, 20-22 hr for B. catenulatum and B. infantis. and 24-25 hr for B. longum and B. breve. No apparent color was observed for B. angulatum and B. adolescentis 28 hr after inoculation. To evaluate the results of pH indicator-based identification, individual isolates were subjected to a colony-PCR experiment with genus-specific primers. The amplified products from the isolates were in good accordance with those from the reference strains at a level of 95% agreement. These results suggest that the present method could be conveniently applied to cell counts, as well as to the preliminary identification of bifidobacteria from a variety of sample types including human feces, dairy products, and commercial probiotic supplements.

Plant Growth Substances Produced by Methylobacterium spp. and Their Effect on Tomato (Lycopersicon esculentum L.) and Red Pepper (Capsicum annuum L.) Growth

  • Ryu, Jeong-Hyun;Madhaiyan, Munusamy;Poonguzhali, Selvaraj;Yim, Woo-Jong;Indiragandhi, Pandiyan;Kim, Kyoung-A;Anandham, Rangasamy;Yun, Jong-Chul;Kim, Kye-Hoon;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1622-1628
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    • 2006
  • Bacteria from the Methylobacterium genus, called pink-pigmented facultative methylotrophic bacteria (PPFMs), are common inhabitants of plants, potentially dominating the phyllosphere population, and are also encountered in the rhizosphere, seeds, and other parts of plants, being versatile in nature. The consistent success of the Methylobacterium plant association relies on methylotrophy, the ability to utilize the one-carbon compound methanol emitted by plants. However, the efficiency of Methylobacterium in plant growth promotion could be better exploited and thus has attracted increasing interest in recent years. Accordingly, the present study investigated the inoculation effects of Methylobacterium sp. strains CBMB20 and CBMB 110 on seed imbibition to tomato and red pepper on the growth and accumulation of phytohormone levels under gnotobiotic conditions. Seeds treated with the Methylobacterium strains showed a significant increase in root length when compared with either the uninoculated control or Methylobacterium extorquens $miaA^-$ knockout mutanttreated seeds. Extracts of the plant samples were used for indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), and dihydrozeatin riboside (DHZR) assays by immunoanalysis. The treatment with Methylobacterium sp. CBMB20 or CBMB 110 produced significant increases in the accumulation of IAA and the cytokinins t-ZR and DHZR in the red pepper extracts, whereas no IAA was detected in the tomato extracts, although the cytokinin concentrations were significantly increased. Therefore, this study proved that the versatility of Methylobacterium as a plant-growth promoting bacteria could be better exploited.

Induced Systemic Resistance by Bacillus vallismortis EXTN-1 Suppressed Bacterial Wilt in Tomato Caused by Ralstonia solanacearum

  • Park, Kyung-Seok;Paul, Diby;Kim, Yong-Ki;Nam, Ki-Woong;Lee, Young-Kee;Choi, Hyo-Won;Lee, Sang-Yeob
    • The Plant Pathology Journal
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    • v.23 no.1
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    • pp.22-25
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    • 2007
  • Biocontrol activity of five strains of selected rhizo-bacteria were tested in tomato against bacterial wilt caused by Ralstonia solanacearum. After root bacterization the plants were grown in a perlite-hydroponic system. Upon challenge inoculation with the pathogen, all of the rhizobacterial strains efficiently suppressed the bacterial wilt in tomato in various rates, at maximum by the strain, Bacillus vallismortis strain EXTN-1. While the percent of infected plants in the non-bacterized control plants were 95%, it was only 65% in plants pre-treated with EXTN-1. It was also demonstrated that the movement of R. solanacearum within the stem was significantly hampered when the plants were root bacterized. As EXTN-1 has no antagonistic properties against R. solanacearum, the bacterial wilt was probably suppressed by a mechanism other than antibiosis. Previously, the strain had been proven to produce an efficient elicitor for inducing systemic resistance in many crops. As the present study confirmed that EXTN-1 has the ability for reducing the pathogen spread in tomato, the strain could be effectively used as a potential biocontrol agent against bacterial wilt.

Comparison of tissue tropism of Newcastle disease vaccine viruses by Immunohistochemistry techniques (면역조직화학기법을 이용한 뉴캣슬병 백신바이러스의 조직친화성 비교)

  • Kim, Min-Jeong;Kwon, Yong-Kuk;Seong, Hwan-Woo;Kang, Shien-Young;Mo, In-Pil
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.539-549
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    • 2004
  • Mean death time of inoculated embryonated egg is one of the methods to determine the virulence of the Newcastle disease viruses (NDV). Evaluation of tissue tropism of NDV in the host has been proposed as an another way to determine the pathogenicity of NDV based on the principal site of viral replication. To evaluate the tissue tropism among NDV, an immunohistochemistry(IHC) technique using monoclonal antibody was applied in one-day-old SPF chickens inoculated with different ND vaccine strains such as Ulster 2C, VG/GA and B1 viruses by eye drop instillation. The tissues used for this comparison were trachea, intestine, Harderian gland and cecal tonsil, which were collected at 0.5, 1, 2, 3, 5, 8, 10, 14 days post inoculation. Among test groups, chickens inoculated with B1 viurs, which is known to replicate in the respiratory system, have IHC positive staining mainly in the trachea and those inoculated with Ulster 2C have IHC positive staining mainly in the intestine. However, chickens inoculated with VG/GA strain have IHC positive staining in both the trachea and intestine. Therefore, a differences in tissue tropism among ND vaccine strains has been proved by the IHC technique. Based on this results, the IHC staining technique could be used to classify the NDV or to study the pathogenesis of NDV in chickens.

Nano-Encapsulation of Plant Growth-Promoting Rhizobacteria and Their Metabolites Using Alginate-Silica Nanoparticles and Carbon Nanotube Improves UCB1 Pistachio Micropropagation

  • Pour, Mojde Moradi;Saberi-Riseh, Roohallah;Mohammadinejad, Reza;Hosseini, Ahmad
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1096-1103
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    • 2019
  • UCB-1 is the commercial rootstock of pistachio. Reproduction of this rootstock by tissue culture is limited by low levels of proliferation rate. Therefore, any compound that improves the proliferation rate and the quality of the shoots can be used in the process of commercial reproduction of this rootstock. Use of plant growth-promoting bacteria is one of the best ideas. Given the beneficial effects of nanoparticles in enhancement of the growth in plant tissue cultures, the aim of the present study was to investigate the effects of nanoencapsulation of plant growth-promoting rhizobacteria (using silica nanoparticles and carbon nanotubes) and their metabolites in improving UCB1 pistachio micropropagation. The experiment was conducted in a completely randomized design with three replications. Before planting, treatments on the DKW medium were added. The results showed that the use of Pseudomonas fluorescens VUPF5 and Bacillus subtilis VRU1 nanocapsules significantly enhanced the root length and proliferation. The nanoformulation of the VUPF5 metabolite led to the highest root length (6.26 cm) and the largest shoot (3.34 cm). Inoculation of explants with the formulation of the metabolites (both bacterial strains) significantly elevated the average shoot length and the fresh weight of plant compared to the control. The explants were dried completely using both bacterial strains directly and with capsule coating after the three days.

Isolation and Characterization of Cold-Adapted PGPB and Their Effect on Plant Growth Promotion

  • Li, Mingyuan;Wang, Jilian;Yao, Tuo;Wang, Zhenlong;Zhang, Huirong;Li, Changning
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1218-1230
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    • 2021
  • Cold-adapted plant growth-promoting bacteria (PGPB) with multiple functions are an important resource for microbial fertilizers with low-temperature application. In this study, culturable cold-adapted PGPB strains with nitrogen fixation and phosphorus solubilization abilities were isolated. They were screened from root and rhizosphere of four dominant grass species in nondegraded alpine grasslands of the Qilian Mountains, China. Their other growth-promoting characteristics, including secretion of indole-3-acetic acid (IAA), production of siderophores and ACC deaminase, and antifungal activity, were further studied by qualitative and quantitative methods. In addition, whether the PGPB strains could still exert plant growth-promoting activity at 4℃ was verified. The results showed that 67 isolates could maintain one or more growth-promoting traits at 4℃, and these isolates were defined as cold-adapted PGPB. They were divided into 8 genera by 16S rRNA gene sequencing and phylogenetic analysis, of which Pseudomonas (64.2%) and Serratia (13.4%) were the common dominant genera, and a few specific genera varied among the plant species. A test-tube culture showed that inoculation of Elymus nutans seedlings with cold-adapted PGPB possessing different functional characteristics had a significant growth-promoting effect under controlled low-temperature conditions, including the development of the roots and aboveground parts. Pearson correlation analysis revealed that different growth-promoting characteristics made different contributions to the development of the roots and aboveground parts. These cold-adapted PGPB can be used as excellent strain resources suitable for the near-natural restoration of degraded alpine grasslands or agriculture stock production in cold areas.