• Journal of Microbiology and Biotechnology
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• v.7 no.3
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• pp.161-166
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• 1997

Nuclei were isolated from the protoplasts of Trichoderma harzianum T95 and treated with colchicine, a polyploid inducer. The nuclei were transferred into the protoplast of multi-auxotrophic Gliocladium virens G88 which cannot grow in minimal medium. The protoplast of G. virens G88 carrying the transferred nuclei were regenerated in a regeneration minimal medium containing $17{\mu}g/ml$ of chloroneb as a haploid inducer. Six intergeneric hybrids between G. virens and T. harzianum were isolated from the regeneration minimal medium. The hybrids could be classified into three types according to morphology, those with an isozyme pattern, those with an protein band and those with an randomly amplified polymorphic DNA(RAPD) pattern produced by random primers and repetitive sequences. The first group was identified to be a haploid recombinant, the second group a heterokaryon, and the third appeared to be petite.

• Journal of Microbiology
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• v.38 no.1
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• pp.1-7
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• 2000

Eight strains of Aeromonas hydrophila isolated from diseased trout in Korea were characterized and compared with an American type strain by various methods including biochemical and physiological tests, PCR, randomly amplified polymorphic DNA (RAPD), plasmid profiling, and gel electrophoresis of total, membrane, and extracellular proteins. Virulence factors such as surface array proteins, cytotoxin, hemolysin, haemagglutinin, and protease were also investigated. The Korean strains showed heterogeneity in Iysine decarboxylase production, utilization of various carbon sources, and production of acetoin. Five strains had the same profiles of total and membrane proteins. Six strains haemagglutinated with trout red blood cells (RBCs) which was inhibited by fucose, galactose, and mannose, except for No. 1 where haemagglutination was inhibited by only galactose and mannose, but not by fucose. Four isolates haemagglutinated with human RBCs which was inhibited by fucose and mannose yet not by galactose. The type strain haemagglutinated only with trout RBCs which was inhibited by fucose, galactose, and mannose. Every isolate secreted protease, hemolysin, cytotoxin, and siderophore, but no enterotoxin. Results showed that the Korean isolates, except for No.7, had very different biochemical and molecular characteristics from those of the American type strain.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.603-607
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• 2004

Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out with a battery of 11 random decamer primers to study band frequency (BF), genetic identity index (I) and mean average percentage difference (MAPD) between Bhadawari and Murrah breeds of buffalo. The primers OPA04 and BG15 resolved a band of 460 bp, which was present only in animals of Bhadawari breed. Whereas, the primers OPA14, BG27 and BG28 produced Murrah specific fragments of sizes 730 bp and 1,230 bp, respectively. The estimate of genetic identity index was highest (0.845) with the primer OPA01 and the lowest (0.479) with the primer BG27. The genetic identity index pooled over the primers was 0.596${\pm}$0.037 between these two breeds. The highest MAPD estimate (53.9) between the two breeds was obtained with the primer BG27 and the lowest (14.3) with the primer OPA01. It might be concluded that the genetic identity index between these two breeds calculated on the basis of BF showed moderate level of genetic identity with the primers employed. MAPD calculated on the basis of uncommon bands also demonstrated lower to medium level of genetic difference between Bhadawari and Murrah breeds of buffalo.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.124-129
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• 2009

In general, the root color of Codonopsis lanceolata is white, but red or blue-colored root is found at a low frequency in nature. Red or blue-colored roots have scarcity value, thus farmers wish to produce colored roots. The factors for determining the color of roots are unclear whether the color is controlled by genetically or simply by environmentally such as soil environment. Using in vitro culture system which is advantageous for setting of the same culture condition, we analyzed the physiological and morphological characteristics and genetic differences among red-, blue- and white lines of C. lanceolata. In the red colored roots, stems of in vitro cultured plantlet were colored in dark red pigment. Histological analysis revealed that the red pigment was accumulated in the outer cortex layer of the stem and determined as anthocyanin. Chlorophyll contents in red root lines were higher than those in white- and blue root lines. Plantlets from red roots were smaller in both shoot length and total leaf area than those from white- and blue roots. Genetic differences among the three different colored C. lanceolata were determined by RAPD (Randomly Amplified Polymorphic DNA) analysis. Each line of colored roots had clear DNA polymorphism. These results indicate that the occurrence of red- and blue colored roots in nature was determined by genetic factors rather than soil enviromental conditions.

• 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.

• Korean Journal of Medicinal Crop Science
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• v.7 no.2
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• pp.94-100
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• 1999

To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.