• Title/Summary/Keyword: species-specificity

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Isolations of the Orchid mycorrhizal Fungi from the Roots of the Korean native Orchids and Inoculations of the Isolates to Four different Orchids (한국 자생란에 난 균근균의 분리와 유묘난에 접종)

  • 이상선;오창호;백기엽;이태수
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.536-542
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    • 1998
  • Ten isolates of the orchid mycorrhizal fungi were isolated from the roots of Korean native orchid plants (Cymbidium goeringii) which inhabitate mainly in southern and western areas of Korea. The growth rates and color of the isolates in potato dextrose agar (PDA) were various. Microscopic observations of the hyphae isolated were identified as Rhizoctonia repens and R. endophytica var endophytica or their related species. R. repens was isolated from the roots of the Korean native orchids, but R. endophytica var endophyica was only isolated from the roots of the commercial orchids introduced from foreign countries. Also, the polymorephic patterns of genomic DNA extracted from selected isolates were compared with those of DNA extracted from the orchid mycorrhizal fungi isolated previously and similar band patterns were observed among those isolates. Five isolates of R. repens were selected and cultured at the oatmeal agar for investigating their symbiosis with orchid plants. The symbiotic specificity between orchid plants and isolated orchid mycorrhizal fungi was observe by growing orchids about six months in the greenhouse. The symbiotic responses of the commercial orchid plants with selected isolates were quite different form different isolates due to the genetic variations.

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Improved Detection of Viable Escherichia coli O157:H7 in Milk by Using Reverse Transcriptase-PCR

  • Choi, Suk-Ho;Lee, Seung-Bae
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.158-165
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    • 2011
  • A sensitive reverse transcriptase-PCR (RT-PCR) method to detect viable Escherichia coli O157:H7 in milk was established. The primer sets were designed based on the nucleotide sequences of the rfbE (per) and wbdN genes in the O157 antigen gene cluster of E. coli O157:H7. RT-PCR using five different primer sets yielded DNA with sizes of 655, 518, 450, and 149-bp, respectively. All five of the E. coli O157:H7 strains were detected by RT-PCR, but 11 other bacterial species were not. The sensitivity of RT-PCR was improved by adding yeast tRNA as a carrier to the crude RNA extract. The RT-PCR amplifying the 149-bp DNA fragment was the most sensitive for detecting E. coli O157:H7 and the most refractory to the bactericidal treatments. Heat treatment at $65^{\circ}C$ for 30 min was the least inhibitory of all bactericidal treatments. Treatment with RNase A strongly inhibited the RT-PCR of heated milk but not unheated milk. This study described RT-PCR methods that are specific and sensitive with a detection limit of 10 E. coli O157:H7 cells, and showed that pre-treating milk samples with RNase A improved the specificity to detect viable bacteria by RT-PCR.

Detection of Vibrio vulnificus by Real-Time PCR targeted to rpoS gene (rpoS 유전자를 대상으로 하는 Real-Time PCR에 의한 Vibrio vulnificus 검출)

  • Kim, Dong-Gyun;Ahn, Sun-Hee;Bae, Ju-Yoon;Kong, In-Soo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.263-266
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    • 2007
  • Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

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Fungal Endophytes from Three Cultivars of Panax ginseng Meyer Cultivated in Korea

  • Park, Sang-Un;Lim, Hyoun-Sub;Park, Kee-Choon;Park, Young-Hwan;Bae, Han-Hong
    • Journal of Ginseng Research
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    • v.36 no.1
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    • pp.107-113
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    • 2012
  • In order to investigate the diversity of endophytes, fungal endophytes in Panax ginseng Meyer cultivated in Korea were isolated and identified using internal transcribed spacer (ITS) sequences of ribosomal DNA. Three cultivars of 3-year-old ginseng roots (Chunpoong, Yunpoong, and Gumpoong) were used to isolate fungal endophytes. Surface sterilized ginseng roots were placed on potato dextrose agar plates supplemented with ampicilin and streptomycin to inhibit bacterial growth. Overall, 38 fungal endophytes were isolated from 12 ginseng roots. According to the sequence analysis of the ITS1-5.8S-ITS2, 38 fungal isolates were classified into 4 different fungal species, which were Phoma radicina, Fusarium oxysporum, Setophoma terrestris and Ascomycota sp. 2-RNK. The most dominant fungal endophyte was P. radicina in 3 cultivars. The percentage of dominant endophytes of P. radicina was 65.8%. The percentage of colonization frequency of P. radicina was 80%, 52.9%, and 75% in Chunpoong, Yunpoong, and Gumpoong, respectively. The second most dominant fungal endophyte was F. oxysporum. The diversity of the fungal endophytes was low and no ginseng cultivar specificity among endophytes was detected in this study. The identified endophytes can be potential fungi for the production of bioactive compounds and control against ginseng pathogens.

Bacteriophage Usage for Bacterial Disease Management and Diagnosis in Plants

  • Vu, Nguyen Trung;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • v.36 no.3
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    • pp.204-217
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    • 2020
  • In nature, plants are always under the threat of pests and diseases. Pathogenic bacteria are one of the major pathogen types to cause diseases in diverse plants, resulting in negative effects on plant growth and crop yield. Chemical bactericides and antibiotics have been used as major approaches for controlling bacterial plant diseases in the field or greenhouse. However, the appearance of resistant bacteria to common antibiotics and bactericides as well as their potential negative effects on environment and human health demands bacteriologists to develop alternative control agents. Bacteriophages, the viruses that can infect and kill only target bacteria very specifically, have been demonstrated as potential agents, which may have no negative effects on environment and human health. Many bacteriophages have been isolated against diverse plant-pathogenic bacteria, and many studies have shown to efficiently manage the disease development in both controlled and open conditions such as greenhouse and field. Moreover, the specificity of bacteriophages to certain bacterial species has been applied to develop detection tools for the diagnosis of plant-pathogenic bacteria. In this paper, we summarize the promising results from greenhouse or field experiments with bacteriophages to manage diseases caused by plant-pathogenic bacteria. In addition, we summarize the usage of bacteriophages for the specific detection of plant-pathogenic bacteria.

Cancer Treatment Using Multiphoton Photodynamic Therapy

  • Zakir Hossain, S.M.;Golam Azam, S.M.;Enayetul Babar, S.M.
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.1-6
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    • 2006
  • Photodynamic therapy (PDT), a newly established treatment for solid tumors, involves the systemic administration of a tumor localizing photosensitizer that is only activated when exposed to light of appropriate wavelength. Photoactivation of photosensitizer in the presence of oxygen results in the formation of highly cytotoxic molecular species, which precipitates necrosis. PDT has now become a promising means for the treatment of cancer due to its specificity, relatively minimal side effects, and inexpensive. However, the application of PDT has been restricted to the treatment of superficial lesions or the use of interstitial light delivery. A single photon generally activates the photochemical reaction in traditional PDT. However the use of multi photon excitation, where two or more photons simultaneously excite a photosensitizer, allows for the use of wavelengths twice as long. Such wavelengths exhibit better transmittance through tissue and thereby deeper penetration is achieved. This paper will review theoretical principles of multi photon excitation, challenges associated with multi photon PDT and update the current and future role of multi photon PDT in cancer.

A comparative study of ectoparasites occurrence between grass carp and silver carp in guilan province culture ponds, Iran

  • Asgharnia, Mehrdad;Ghasemi, Mohaddes
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.169-176
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    • 2021
  • Parasitic infection is among the most common problems for carp cultivation. They are also important for the principal entrance of other hazardous infections as well. This study was carried out for determining of parasitic fauna of two major carp known as silver and grass carp with the comparison of prevalence value and intensity rate of parasites among them, alongside the relationship between the biometric characteristics and host sex with the infection level. For this purpose, a total of 94 fish samples were caught randomly using a fishing net, from Guilan ponds during spring and summer of the year 2018 and transported alive to the laboratory. Upon arriving, the biometric characteristics and genus of each carp were measured individually. Specimens were then acquired from the skin, gills, and eyes of the carp and examined according to standard parasitology methods. Recovered parasites were observed under a light microscope and then fixed for identification. As the result, the occurrence and intensity in the higher length group were comparatively greater than the lower one. Also, the prevalence and intensity of total parasites in male carp were higher than in females. In this research, Dactylogyrus hypophthalmichthys and Dactylogyrus aristhichtys were observed in silver carp and Dactylogyrus lamellatus was detected in grass carp. In the paper below, we found that the host specificity varies in different species of Dactylogyrus isolated from grass carp and silver carp.

Strain-specific Detection of Kimchi Starter Leuconostoc mesenteroides WiKim33 using Multiplex PCR

  • Lee, Moeun;Song, Jung Hee;Park, Ji Min;Chang, Ji Yoon
    • Journal of the Korean Society of Food Culture
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    • v.34 no.2
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    • pp.208-216
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    • 2019
  • Leuconostoc spp. are generally utilized as kimchi starters, because these strains are expected to have beneficial effects on kimchi fermentation, including improvement of sensory characteristics. Here, we developed a detection method for verifying the presence of the kimchi starter Leuconostoc mesenteroides WiKim33, which is used for control of kimchi fermentation. A primer set for multiplex polymerase chain reaction was designed based on the nucleotide sequence of the plasmids in strain WiKim33, and their specificity was validated against 45 different strains of Leuconostoc spp. and 30 other strains. Furthermore, the starter strain consistently tested positive, regardless of the presence of other bacterial species in starter kimchi during the fermentation period. Our findings showed that application of a strain-specific primer set for strain WiKim33 presented a rapid, sensitive, and specific method for detection of this kimchi starter strain during natural kimchi fermentation.

Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.155-159
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    • 2021
  • This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Flavonoids Biotransformation by Human Gut Bacterium Dorea sp. MRG-IFC3 Cell-Free Extract

  • Huynh Thi Ngoc Mi;Heji Kim;Jong Suk Lee;Bekir Engin Eser;Jaehong Han
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1270-1275
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    • 2024
  • Human gut bacterium Dorea sp. MRG-IFC3 is unique in that it is capable of metabolizing puerarin, an isoflavone C-glycoside, whereas it shows broad substrate glycosidase activity for the various flavonoid O-glycosides. To address the question on the substrate specificity, as well as biochemical characteristics, cell-free biotransformation of flavonoid glycosides was performed under various conditions. The results showed that there are two different enzyme systems responsible for the metabolism of flavonoid C-glycosides and O-glycosides in the MRG-IFC3 strain. The system responsible for the conversion of puerarin was inducible and comprised of two enzymes. One enzyme oxidizes puerarin to 3"-oxo-puerarin and the other enzyme converts 3"-oxo-puearin to daidzein. The second enzyme was only active toward 3"-oxo-puerarin. The activity of puerarin conversion to daidzein was enhanced in the presence of Mn2+ and NAD+. It was concluded that the puerarin C-deglycosylation by Dorea sp. MRG-IFC3 possibly adopts the same biochemical mechanism as the strain PUE, a species of Dorea longicatena.