• Title/Summary/Keyword: 185 rDNA

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Leaf variants of Pinus and their ITS DNA sequences (소나무속 잎 변이와 그의 ITS DNA 염기서열)

  • Koo, JaChoon;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.43 no.1
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    • pp.63-68
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    • 2013
  • ITS DNA sequences of two variants of Pinus spp. having single fasciculate leaf or two to three fasciculate leaves within an individual were analysed in order to determine their origin. Also, the same DNA locus of P. densiflora, P. rigida and P. koraiensis, distributed at the same region together with the OTUs having leaf variations, were analysed to compare with each other. Aligned sequences including ITS1, 5.8S and ITS2 were 580~584 bp in length. The 5.8S region was well conserved among all the OTUs we tested except for P. koraiensis, which has two nucleotide substitutions. The partial ITS1 region upstream of the 5.8S region showed relatively high sequence diversity compare to the ITS2 and has 181~185 bp in length. In this region, the sequences of the two variants were identical to that of P. densiflora. The ITS2 has identical for all OTUs in length and the two variants also have same sequences compare to that of P. densiflora. These two variants and samples of P. densiflora were grouped together in the UPGMA tree with 100% similarity level. The result strongly suggest that these two variants were originated from P. densiflora.

Identification of Yeasts Isolated from Wild Flowers Collected in Coast Areas of Korea Based on the 26S rDNA Sequences (우리나라 일부 해안 지역 야생화들로부터 분리한 효모들의 분자 생물학적 동정)

  • Min, Jin Hong;Lee, Hyang Burm;Lee, Jong Soo;Kim, Ha Kun
    • The Korean Journal of Mycology
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    • v.41 no.3
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    • pp.185-191
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    • 2013
  • Several yeast colonies were isolated from wild flowers collected from East, West and South coast areas of Korea by plating of flower suspensions on the YPD plates containing antibiotics, streptomycin and ampicillin. Polymerase chain reactions (PCR) were performed for the amplification of D1/D2 region of 26S rDNA for those colonies. PCR-amplified nucleotide sequences were compared using BLAST for their identification. As results, 27 yeast strains belonged to 15 species were isolated from wild flowers collected at Donghae, where is located in eastern coast of Korea. Also, 34 strains belonged to 17 species were isolated from wild flowers of Daecheon, where is located in western coast of Korea. In addition, 22 strains belonged to 13 species were isolated from wild flowers collected at Wando, where is located in southern coast of Korea. Among those 45 species isolated from 3 different collection sites, only 4 species including Cryptococcus laurentii, Metschnikowia koreensis, Pseudozyma rugulosa, and Rhodotorula mucilaginosa were found from all 3 different collection sites. And 5 species including Cryptococcus aureus, Cryptococcus flavus, Hanseniaspora uvarum, Pichia guilliermondii, and Rhodosporidium fluviale were overlapped from the at least 2 different collection sites. Other 23 species were found only in a specific collection sites implying that each area has distinctive yeast flora.

Genetic analysis of clubroot resistance in Chinese cabbage using single spore isolate of Plasmodiophora brassicae and development of RAPD marker linked to its resistance gene

  • Cho, Kwang-Soo;Hong, Su-Young;Han, Young-Han;Yoon, Bong-Kyeong;Ryu, Seoung-Ryeol;Woo, Jong-Gyu
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.101-106
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    • 2008
  • To identify inheritance of clubroot disease resistance genes in Chinese cabbage, seedling tests of $BC_1P_1,\;BC_1P_2$, and $F_2$ populations derived from $F_1$ hybrid(var. CR Saerona) using single spore isolate(race 4 identified with William's differential host) from Plasmodiophora brassciae were conducted. Resistance(R) and susceptible(S) plants segregated to 1:0 in backcross to the resistant parent. The $F_2$ population segregated in a 3(R):1(S) ratio. This result implied that the resistance of clubroot disease is controlled by a single dominant gene to the race 4 of P. brassicae in CR Saerona. To develop DNA markers linked to clubroot resistance genes, 185 plants of CR Saerona among $F_2$ populations were used. A total of 300 arbitrary decamer was applied to $F_2$ population using BSARAPD(Bulked segregant analysis-Randomly amplified polymorphic DNA). One RAPD marker linked to clubroot resistance gene in CR Saerona($OPJ_{1100}$) was identified. This marker was 3.1 cM in distance from resistance gene in $F_2$ population. This marker may be useful for a marker-assisted selection(MAS) and gene pyramiding of the clubroot disease resistant gene in Chinese cabbage breeding programs.

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Antioxidant Capacity of Crude Extract and Its Solvent Fractions of Arctic Terrestrial Plant Ranunculus heperporeus (북극식물 Ranunculus hyperboreus의 추출물과 용매분획물의 항산화 활성)

  • Lee, Jung Im;Kim, Hojun;Seo, Hyo-Won;Kong, Chang-Suk;Seo, Youngwan
    • Ocean and Polar Research
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    • v.38 no.3
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    • pp.185-193
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    • 2016
  • In this study, antioxidative potentials of the crude extract and its four solvent fractions from the Arctic terrestrial plant Ranunculus heperporeus were evaluated by using four different activity tests, including the inhibition of intracellular reactive oxygen species (ROS) and lipid peroxidation in Raw 264.7 cells as well as determining the extent of both the scavenging of peroxynitrite ($ONOO^-$) and the oxidative damage of genomic DNA purified from Raw 264.7 cells. Based on a comparative analysis, n-BuOH, and 85% aq.MeOH solvent fractions showed good scavenging effects on the production of intracellcular ROS and inhibited membrane lipid peroxidation and DNA oxidation. In addition, n-BuOH and 85% aq.MeOH fractions exhibited good scavenging effects on both authentic peroxynitrite and one generated from SIN-1. Among the samples tested, the n-BuOH fraction revealed the strongest antioxidant effect.

Isolation and development of Bacillus subtilis S1-0210 as a biocontrol agent of gray mold of strawberry

  • Nguyen, Hang T.T.;Oh, S.O.;Hur, J.S.;Koh, Y.J.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.98.1-98
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    • 2003
  • Antagonistic effect of bacterial strains isolated from phylloplane of strawberry plants grown In greenhouse was tested on Botrytis cinerea Among the promising bacterial strains, Bacillus sp. S1-0210 showed highest inhibition of mycelial growth of B. cinerea and a broad spectrum of antifungal activities against many plant pathogenic fungi in vitro. Bacillus sp. S1-0210 was identified as Bacillus subtilis based on the analysis of 185 rDNA as well as its biochemical characteristics. Application of wettable powder formulation of B. subtiiis S1-0210 significantly reduced the incidence of gray mold on trawberry fruits during storage. Results showed that treatment of B. subtilis S1-0210 decreased the incidence of gray mold by 4.8% whereas the incidence in control was 77.9%, indicating that the formulation of B. subtilis S1-0210 will be practically applied on strawberry fruits as a biocontrol agent of gray mold during storage.

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Monitoring of Microorganisms Added into Oil-Contaminated Microenvironments by Terminal-Restriction Fragment Length Polymorphism Analysis

  • JUNG SEONG-YOUNG;LEE JUNG-HYUN;CHAI YOUNG-GYU;KIM SANG-JIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1170-1177
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    • 2005
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

Cloning and Characterization of a Gene Cluster for Cyclohexanone Oxidation in Rhodococcus sp. TK6

  • Choi Jun-Ho;Kim Tae-Kang;Kim Young-Mog;Kim Won-Chan;Park Kunbawui;Rhee In-Koo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.511-518
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    • 2006
  • A gene cluster for cyclohexanone oxidation was cloned from Rhodococcus sp. TK6, which is capable of growth on cyclohexanone as the sole carbon source. The 9,185-bp DNA sequence analysis revealed seven potential open reading frames (ORFs), designated as ssd-chnR-chnD-chnC-chnB-chnE-partial pcd. The chnBCDE genes encode enzymes for the four-step conversion of cyclohexanone to adipic acid, catalyzed by cyclohexanone monooxygenase (ChnB), $\varepsilon-caprolactone$ hydrolase (ChnC), 6-hydroxyhexanoate dehydrogenase (ChnD), and 6-oxohexanoate dehydrogenase (ChnE). Furthermore, the presence of a regulatory element in the downstream region of the chnD gene supports the notion that chnR is a putative regulatory gene. Among them, the activity of ChnB was confirmed and characterized, following their expression and purification in Escherichia coli harboring the modified chnB gene (chnB gene with 6 successive codons for His at the 3' terminus).

Isolation and Characterization of Surfactin-producing Bacillus amyloliquefaciens YJ07 from Spent Mushroom (Pleurotus eryngii) Substrates (새송이버섯 수확 후 배지로부터 surfactin 생성 Bacillus amyloliquefaciens YJ07의 분리 및 특성)

  • Shin, Pyung Gyun;Yoo, Young Bok;Cho, Yong Un;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.180-185
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    • 2011
  • Spent mushroom substrates (SMS) is a by-product remaining after a crop of mushrooms. About four surfactin-producing strains were isolated from SMS (Pleurotus eryngii). Among of them, one isolate, which designated to YJ07, potentially showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium. The biochemical characteristics of the strain YJ07 was similar with Bacillus subtilis and Bacillus amyloliquefaciens by Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis of the strain YJ07 also showed that the strain YJ07 was most closely related to Bacillus amyloliquefaciens with sequence similarity of 99.5%. On the basis of their biochemical characteristics and phylogenetic distinctiveness, the strain YJ07 was classified within the genus Bacillus as Bacillus amyloliquefaciens YJ07. The antifungal compound from B. amyloliquefaciens YJ07 was similar to lipopeptide surfactin from Bacillus subtilis by TLC and HPLC analysis.

Fungal Diversity in Composting Process of Pig Manure and Mushroom Cultural Waste Based on Partial Sequence of Large Subunit rRNA

  • Cho, Kye-Man;Kwon, Eun-Ju;Kim, Sung-Kyum;Kambiranda, Devaiah M;Math, Reukaradhya K;Lee, Young-Han;Kim, Jung-Ho;Yun, Han-Dae;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.743-748
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    • 2009
  • Fungal diversity during composting was investigated by culture-independent rDNA sequence analysis. Composting was carried out with pig manure and mushroom cultural waste using a field-scale composter (Hazaka system), and samples were collected at various stages. Based on partial sequence analysis of large subunit (LSU) ribosomal RNA (rRNA) and sequence identity values, a total of 12 different fungal species were found at six sampling sites; Geotrichum sp., Debaryomyces hansenii, Monographella nivalis, Acremonium strictum, Acremonium alternatum, Cladosporium sphaerospermum, Myriangium durosai, Pleurotus eryngii, Malassezia globosa, Malassezia restricta, Rhodotorula glutinis, and Fusarium sporotrichioides. Geotrichum sp. of the class Saccharomycetes was the most predominant fungal species throughout the composting process (185 out of a total of 236 identified clones, or 78.4%), followed by Acremonium strictum (7.6%), Monographella nivalis (5.1%), and Pleurotus eryngii (3.8%). The prevalence of Geotrichum sp. was the lowest (61.1%) at the beginning of composting, and then gradually increased to 92.5% after 10 days of composting.

Identification of a Newly Isolated Protease-producing Bacterium, Bacillus subtilis FBL-1, from Soil (토양으로부터 새로이 분리된 단백질 분해효소 생산 미생물 Bacillus subtilis FBL-1의 동정)

  • Kim, Mina;Si, Jin-Beom;Wee, Young-Jung
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.185-193
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    • 2016
  • A novel proteolytic bacterium was isolated from soil at Yeungnam University, South Korea. The strain, named FBL-1, was rod-shaped with a smooth surface. Biolog and API 50CHB test results revealed that strain FBL-1 was a Bacillus species. Based on 16S rDNA sequencing and chemotaxonomic characterization, the strain was identified as Bacillus subtilis because it had the highest homology with Bacillus subtilis subsp. subtilis NCIB 3610 (99.5%). In liquid culture at 37℃ with shaking at 200 rpm, fructose and yeast extract were found to be the best carbon and nitrogen sources, respectively, for cell growth and protease production. The highest protease activity (451.640 U/ml) was obtained when the strain was cultured in medium containing 20 g/l of fructose and 5 g/l of yeast extract. Although further studies are needed to characterize the protease and enhance its activity, the newly isolated protein-degrading B. subtilis FBL-1 can be applicable for the production of peptides and for the degradation of proteins in various industries.