• Title/Summary/Keyword: RAPD

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Totipotential, Morphological, Biochemical Comparisons between Nonembryogenic Callus and Embryogenic Callus in Water Dropwort(Oenanthe stolonifera DC) (미나리에서 비배발생캘러스와 배발생캘러스간의 분화능력 및 해부학적, 생화학적 특성비교)

  • 빈철구;김병동
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.167-173
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    • 1997
  • The embryogenic callus (EC), from which somatic embryos could be induced, was compared with nonembryogenic callus(NE) to study the origin and features of totipotent cell in water dropwort (Oenanthe stolonifera DC). To induce and maintain of EC and the NE, meristematic stem and immature floret were inoculated in MS media supplemented with 1 mg/L 2,4-D, and with 2.5 mg/L NAA and 5mg/L BA, respectively, The EC was not induced from the NE even after subculturing in MS medium supplemented with 1 mg/L 2,4-D. Plantlets were not regenerated from the NE in hormone-free medium. In histochemical comparison of the EC with the NE by light microscopy, the EC had smaller cells in size, dense cytoplasm, and more starch granules of cells compared to the NE cells. The cell from the EC, as observed by transmission electron microscopy, had smaller vaculoes, well developed ribosomes, mitochondria, and endoplasmic reticulum, whereas the cells from the NE had larger vacuoles and underdeveloped organelles. In protein pattern from NE, EC and Somatic embryo (SE), as analyzed by SDS polyacrylamide gel electrophoresis, different proteins specific for tissue were observed: 17 and 28 KD for NE, 50, 52, 57, 66, 68 KD for EC and 20 KD for SE. DNA polymorphism was also observed between EC and NE as analyzed by RAPD (randomly amplified polymorphic DNA) method. The origin of totipotent stem cell and the relationship between irreversible genomic change arose in differentiation and the loss of totipotency in plant were discussed.

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Ginseng Conservation Program in Russian Primorye: Genetic Structure of Natural and Cultivated Populations

  • Yu.N. Zhuravlev;O.G. Koren;G.D. Reunova;E.V Artyukova;M.M. Kozyrenko;T.I. Muzarok;I.L. Kats
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.509-521
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    • 2002
  • 'The Regional complex long-term program of restoration (reintroduction) of Primoryes ginseng population up to 2005' elaborated by Primorye governor administration, Regional Committee of Natural Resources and Russian Academy of Sciences operates in Russian Primorye. The Institute of Biology and Soil Science (IBSS) provides the scientific implementation of this program including the genetic analysis of extant ginseng populations, plant reproduction and offspring identification. According to our investigations, the genetic resource of P. ginseng in Primorye is represented by three populations of wild-growing ginseng and a few private plantations. The results obtained by RAPD allowed concluding that this resource is dispersed among the wild and cultivated ginseng sub-populations in such a way that each of sub-populations studied has to be represented in living plant collection as a stock material to maintain species genetic variability. The allozyme analyses also showed that the small sub-populations of natural ginseng are characterized by unique genetic diversity and, therefore, they all need to be represented in reintroduction centers. Additionally the allozyme analysis discovered that the Blue Mountain and Khasan populations possess the most genetic diversity. So, at least one more reproductive ginseng unit has to be created besides two already existing reintroduction centers representing the Sikhote-Alin and the Blue Mountain populations.

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Small-scale spatial genetic structure of Asarum sieboldii metapopulation in a valley

  • Jeong, Hyeon Jin;Kim, Jae Geun
    • Journal of Ecology and Environment
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    • v.45 no.3
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    • pp.97-104
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    • 2021
  • Background: Asarum sieboldii Miq., a species of forest understory vegetation, is an herbaceous perennial belonging to the family Aristolochiaceae. The metapopulation of A. sieboldii is distributed sparsely and has a short seed dispersal distance by ants as their seed distributor. It is known that many flowers of A. sieboldii depend on self-fertilization. Because these characteristics can affect negatively in genetic structure, investigating habitat structure and assessment of genetic structure is needed. A total of 27 individuals in a valley were sampled for measuring genetic diversity, genetic distance, and genetic differentiation by RAPDPCR. Results: The habitat areas of A. sieboldii metapopulation were relatively small (3.78~33.60 m2) and population density was very low (five to seven individuals in 20×20 m quadrat). The habitat of A. sieboldii was a very shady (relative light intensity = 0.9%) and mature forest with a high evenness value (J = 0.81~0.99) and a low dominance value (D = 0.19~0.28). The total genetic diversity of A. sieboldii was quite high (h = 0.338, I = 0.506). A total of 33 band loci were observed in five selected primers, and 31 band loci (94%) were polymorphic. However, genetic differentiation along the valley was highly progressed (Gst = 0.548, Nm = 0.412). The average genetic distance between subpopulations was 0.387. The results of AMOVA showed 52.77% of variance occurs among populations, which is evidence of population structuring. Conclusions: It is expected that a small-scale founder effect had occurred, an individual spread far from the original subpopulation formed a new subpopulation. However, geographical distance between individuals would have been far and genetic flow occurred only within each subpopulation because of the low density of population. This made significant genetic distance between the original and new population by distance. Although genetic diversity of A. sieboldii metapopulation is not as low as concerned, the subpopulation of A. sieboldii can disappear by stochastic events due to small subpopulation size and low density of population. To prevent genetic isolation and to enhance the stable population size, conservative efforts such as increasing the size of each subpopulation or the connection between subpopulations are needed.

Biotransformation of Reactive Red 141 by Paenibacillus terrigena KKW2-005 and Examination of Product Toxicity

  • Sompark, Chalermwoot;Singkhonrat, Jirada;Sakkayawong, Niramol
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.967-977
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    • 2021
  • A total of 37 bacterial isolates were obtained from dye-contaminated soil samples at a textile processing factory in Nakhon Ratchasima Province, Thailand, and the potential of the isolates to decolorize and biotransform azo dye Reactive Red 141 (RR141) was investigated. The most potent bacterium was identified as Paenibacillus terrigena KKW2-005, which showed the ability to decolorize 96.45% of RR141 (50 mg/l) within 20 h under static conditions at pH 8.0 and a broad temperature range of 30-40℃. The biotransformation products were analyzed by using UV-Vis spectrophotometry and Fourier-transform infrared spectroscopy. Gas chromatography-mass spectroscopy analysis revealed four metabolites generated from the reductive biodegradation, namely sodium 3-diazenylnaphthalene-1,5-disulfonate (I), sodium naphthalene-2-sufonate (II), 4-chloro-1,3,5-triazin-2-amine (III) and N1-(1,3,5-triazin-2-yl) benzene-1,4-diamine (IV). Decolorization intermediates reduced phytotoxicity as compared with the untreated dye. However, they had phytotoxicity when compared with control, probably due to naphthalene and triazine derivatives. Moreover, genotoxicity testing by high annealing temperature-random amplified polymorphic DNA technique exhibited different DNA polymorphism bands in seedlings exposed to the metabolites. They compared to the bands found in seedlings subjected to the untreated dye or distilled water. The data from this study provide evidence that the biodegradation of Reactive Red 141 by P. terrigena KKW2-005 was genotoxic to the DNA seedlings.

Isolation of 2 Bacillus Strains with Strong Fibrinolytic Activities from Kimchi

  • Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Afifah, Diana Nur;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.439-446
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    • 2020
  • Two Bacillus strains, K3 and K208, both demonstrating strong fibrinolytic activities were isolated from Kimchi, a traditional Korean preparation of fermented vegetables. Isolates were subjected to various molecular biology based identification methods including RAPD-PCR and identified as B. subtilis and B. velezensis, respectively. Tryptic soy broth (TSB) was found to best maintain both the growth and the fibrinolytic activity of these strains. Culture supernatants were analyzed by SDS-PAGE and fibrin zymography, and the results indicate that a 40 and 27 kDa band seem to be responsible for the fibrinolytic activities of these two isolates and the 27 kDa band was subsequently identified as the mature form of AprE, the major fibrinolytic enzyme. Thus the aprE genes were cloned and the translated amino acid sequences demonstrated 99.3% identity with each other, and 86.5% identity with BsfA, a fibrinolytic enzyme from B. subtilis ZA400 also isolated from Kimchi, and AprE2, a fibrinolytic enzyme from B. subtilis CH3-5 isolated from Cheonggukjang, a traditional Korean fermented soy. Given this B. subtilis K3 and B. velezensis K208 may be promising starter cultures in the production of fermented foods.

Molecular methods for diagnosis of microbial pathogens in muga silkworm, Antheraea assamensis Helfer (Lepidoptera: Saturniidae)

  • Gangavarapu Subrahmanyam;Kangayam M. Ponnuvel;Kallare P Arunkumar;Kamidi Rahul;S. Manthira Moorthy;Vankadara Sivaprasad
    • International Journal of Industrial Entomology and Biomaterials
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    • v.47 no.1
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    • pp.1-11
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    • 2023
  • The Indian golden muga silkworm, Antheraea assamensis Helfer is an economically important wild silkworm endemic to Northeastern part of India. In recent years, climate change has posed a threat to muga silk production due to the requirement that larvae be reared outdoors. Since the muga silkworm larvae are exposed to the vagaries of nature, the changing climate has increased the incidence of microbial diseases in the rearing fields. Accurate diagnosis of the disease causing pathogens and its associated epidemiology are prerequisites to manage the diseases in the rearing field. Although conventional microbial culturing methods are widely used to identify pathogenic bacteria, they would not provide meaningful information on a wide variety of silkworm pathogens. The information on use of molecular diagnostic tools in detection of microbial pathogens of wild silk moths is very limited. A wide range of molecular and immunodiagnostic techniques including denaturing gradient gel electrophoresis (DGGE), random amplified polymorphism (RAPD), 16S rRNA/ITSA gene sequencing, multiplex polymerase chain reaction (M-PCR), fluorescence in situ hybridization (FISH), immunofluorescence, and repetitive-element PCR (Rep-PCR), have been used for detecting and characterizing the pathogens of insects with economic significance. Nevertheless, the application of these molecular tools for detecting and typing entomopathogens in surveillance studies of muga silkworm rearing is very limited. Here, we discuss the possible application of these molecular techniques, their advantages and major limitations. These methods show promise in better management of diseases in muga ecosystem.

Molecular Identification and Effects of Temperature on Survival and Growth of Hybrids between Haliotis gigantea Gmelin (♀) and Haliotis discus hannai Reeve (♂)

  • An, Hye Suck;Han, Jong Won;Hwang, Hyun-Ju;Jeon, Hancheol;Jung, Seung-Hyun;Jo, Seonmi;Choi, Tae-Young;Hyun, Young Se;Song, Ha Yeun;Whang, Ilson
    • Journal of Marine Life Science
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    • v.2 no.2
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    • pp.83-89
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    • 2017
  • In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.

Antimicrobial Resistance of Seventy Lactic Acid Bacteria Isolated from Commercial Probiotics in Korea

  • Eunju Shin;Jennifer Jaemin Paek;Yeonhee Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.500-510
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    • 2023
  • In this study, lactic acid bacteria were isolated from 21 top-selling probiotic products on Korean market and their antimicrobial resistance were analyzed. A total 152 strains were claimed to be contained in these products and 70 isolates belonging to three genera (Bifidobacterium, Lactobacillus, and Lactococcus) were obtained from these products. RAPD-PCR showed diversity among isolates of the same species except for two isolates of Lacticaibacillus rhamnosus from two different products. The agar dilution method and the broth dilution method produced different MICs for several antimicrobials. With the agar dilution method, five isolates (three isolates of Bifidobacterium animalis subsp. lactis, one isolate of B. breve, one isolate of B. longum) were susceptible to all nine antimicrobials and 15 isolates were multi-drug resistant. With the broth microdilution method, only two isolates (one isolate of B. breve and one isolate of B. longum) were susceptible while 16 isolates were multi-drug resistant. In this study, only two AMR genes were detected: 1) lnu(A) in one isolate of clindamycin-susceptible and lincomycin-resistant Limosilactobacillus reuteri; and 2) tet(W) in one tetracycline-susceptible isolate of B. longum B1-1 and two tetracycline-susceptible isolates and three tetracycline resistant isolates of B. animalis subsp. lactis. Transfer of these two genes via conjugation with a filter mating technique was not observed. These results suggest a need to monitor antimicrobial resistance in newly registered probiotics as well as probiotics with a long history of use.

Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

Clinical aspects of an outbreak of Serratia marcescens infections in neonates (신생아 중환자실에서 Serratia marcescens에 의한 집단 감염 발생에 대한 고찰)

  • Sung, Min-Jung;Chang, Chul-Hun;Yoon, Yeon-Kyong;Park, Su-Eun
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.500-506
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    • 2006
  • Purpose : We evaluated an outbreak of Serratia marcescens infections in 24 neonates in a neonatal intensive care unit(NICU). Methods : From January to August, 2004 a nosocomial outbreak of S. marcescens occurred in our NICU. We describe the clinical characteristics of the outbreak and analyse the risk factors for infections with S. marcescens. After the outbreak stopped, 7 isolates from blood were typed using rapid amplified polymorphic DNA analysis(RAPD). Results : S. marcescens was isolated from 24 neonates, 19 infected and 5 colonized. Seven out of nineteen neonates had bacteremia, 4 had ventilator associated pneumonia, 4 had purulent conjunctivitis, 2 had UTI, 1 had meningitis and 1 had a wound infection. Three neonates died due to S. marcescens infection, 2 of 3 had ventilator associated pneumonia, 1 had meningitis complicated with abscess. The mortality rate of S. marcescens infection was 15.8%. Factors associated with S. marcescens infections were previous antibiotic therapy, indwelling catheter and use of ventilators. The isolated strains were resistant to most antibiotics, but frequently sensitive to imipenem, bactrim and amikacin. RAPD typing results show that at least 3 epidemic strains were related with this outbreak. But one genotype was predominant type in this outbreak. The control measures were instituted and the outbreak stopped within 2 months. Conclusion : S. marcescens can cause rapidly spreading outbreaks associated with fatal infections in neonates. If S. marcescens is isolated from clinical specimens, meticulous infection control measures and epidemiologic investigations should be done at an early stage of the outbreak.