• Title/Summary/Keyword: sequence diversity

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Isolation and Characterization of Fungal Diversity from Crop Field Soils of Nigeria

  • Yadav, Dil Raj;Kim, Sang Woo;Adhikari, Mahesh;Babu, Anam Giridhar;Um, Yong Hyun;Gim, Eun Bi;Yang, Jae Seok;Lee, Hyug Goo;Lee, Youn Su
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.49-49
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    • 2014
  • In order to find indigenous beneficial fungal species from crop field soils of Nigeria, 23 soil samples were collected from various places of Nigeria in June, 2013 and fungi were isolated through serial dilution technique. Isolated fungi were purified and differentiated according to their morphological and microscopic characteristics. In total, 38 different representative isolates were recovered and the genomic DNA of each isolates was extracted using QIAGEN$^{(R)}$ Plasmid Mini Kit (QIAGEN Sciences, USA) and the identification of fungi was carried out by sequence analysis of internal transcribed spacer (ITS) region of the 18S ribosomal DNA (18S rDNA). Recovered isolates belonged to 9 fungal genera comprising Fusarium, Aspergillus, Chaetomium, Coniothyrium, Dipodascaceae, Myrothecium, Neosartorya, Penicillium and Trichoderma. Aspergillus spp., Penicillium spp. and Trichoderma spp. were the most dominant taxa in this study. The antagonistic potentiality of species belonged to Trichoderma against 10 phytopathogenic fungi (F. oxysporum, C. gloesporoides, P. cytrophthora, A. alternata, A. solani, S. rolfsii, F. solani, R. solani, S. sclerotiorum and P. nicotiana) was assessed in vitro using dual culture assay. The dual culture assay results showed varied degree of antagonism against the tested phytopathogens. The potential Trichoderma spp. will be further evaluated for their antagonistic and plant growth promotion potentiality under in vivo conditions.

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Analysis of Genetic Diversity in Echinochloa Species Using Random Amplified Polymorphic DNAs(RAPDs) Markers (RAPD Marker를 이용한 피 수집종의 유연관계 분석)

  • Kim, Kil-Ung;Sohn, Jae-Keun;Shin, Dong-Hyun;Kim, Kyung-Min;Kim, Hak-Yoon;Lee, In-Jung
    • Korean Journal of Weed Science
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    • v.18 no.1
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    • pp.76-83
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    • 1998
  • Echinochloa species maintained by selling for more than 10 years were classified using random amplified polymorphic DNAs(RAPDs) analysis. Seventy-four decamer of randomly sequence markers were used to classify intraspecific variation irt Echinochloa species. The number of amplification products increased with increasing GC content of the primer in the range between 60% and 70% GC. Single-base substitutions of a primer altered amplification, providing new polymorphisms. The size of amplified DNA was mostly between 0.40kbp and 1.4kbp with the most common bands at 1.1kbp. Echinochloa species were detected with 6 primers which generated 26 polymorphic amplified DNAs. By hierarchical cluster analysis, Echinochloa species collected in Korea were divided into three groups. These results revealed that RAPD markers are useful tools for the determination of genetic variations in Echinochloa species.

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Molecular Profiling of Rhizosphere Bacterial Communities Associated with Prosopis juliflora and Parthenium hysterophorus

  • Jothibasu, K.;Chinnadurai, C.;Sundaram, S.P.;Kumar, K.;Balachandar, D.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.301-310
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    • 2012
  • Prosopis juliflora and Parthenium hysterophorus are the two arid, exotic weeds of India that are characterized by distinct, profuse growth even in nutritionally poor soils and environmentally stressed conditions. Owing to the exceptional growth nature of these two plants, they are believed to harbor some novel bacterial communities with wide adaptability in their rhizosphere. Hence, in the present study, the bacterial communities associated with the rhizosphere of Prosopis and Parthenium were characterized by clonal 16S rRNA gene sequence analysis. The culturable microbial counts in the rhizosphere of these two plants were higher than bulk soils, possibly influenced by the root exudates of these two plants. The phylogenetic analysis of V1_V2 domains of the 16S rRNA gene indicated a wider range of bacterial communities present in the rhizosphere of these two plants than in bulk soils and the predominant genera included Acidobacteria, Gammaproteobacteria, and Bacteriodetes in the rhizosphere of Prosopis, and Acidobacteria, Betaproteobacteria, and Nitrospirae in the Parthenium rhizosphere. The diversity of bacterial communities was more pronounced in the Parthenium rhizosphere than in the Prosopis rhizosphere. This culture-independent bacterial analysis offered extensive possibilities of unraveling novel microbes in the rhizospheres of Prosopis and Parthenium with genes for diverse functions, which could be exploited for nutrient transformation and stress tolerance in cultivated crops.

Diversity of Fungal Endophytes in Various Tissues of Panax ginseng Meyer Cultivated in Korea

  • Park, Young-Hwan;Lee, Soon-Gu;Ahn, Doek-Jong;Kwon, Tae-Ryong;Park, Sang-Un;Lim, Hyoun-Sub;Bae, Han-Hong
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.211-217
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    • 2012
  • Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

Microsatellite Analysis of Silkworm Strains (Bombyx mori) of Japan Origin Preserved in Korea

  • Kim, Kee Young;Kang, Pil Don;Kim, Mi Ja;Ryu, Kang Sun;Park, Jeong Sun;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.28 no.2
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    • pp.39-50
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    • 2014
  • In order to understand the diversity and genetic relationships of silkworm strains preserved in Korea, we genotyped 78 Bombyx mori strains (Bombycidae: Lepidoptera) originating from Japan, using eight polymorphic microsatellite loci. We obtained per-locus allele numbers ranging from 5 to 16 (with an average value of 9.1), per-locus observed heterozygosity ranging from 0.13 to 1.00, and per-locus polymorphic information content ranging from 0.36 to 0.77, indicating that some loci are highly variable. Phylogenetic analysis with the eight concatenated microsatellite loci showed no clustering based on known strain characteristics and origin. Nineteen strain-specific apomorphic alleles, which discriminated 16 of the 78 silkworm strains, were obtained from eight loci. These strain-specific alleles can thus be utilized for routine discrimination of strains from Japan, without any further typing of other loci. Homozygotes were also observed at some loci (27 of 118 genotypes), which can also be used to discriminate several strains by typing a few loci. These results showed that eight microsatellite loci described herein were sufficiently variable to discriminate among the 78 silkworm strains we examined, and may be useful for future investigations of this economically important species.

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

  • Alam, Nuhu;Kim, Jeong-Hwa;Shim, Mi-Ja;Lee, U-Youn;Lee, Tae-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.89-96
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    • 2010
  • This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at $25^{\circ}C$ and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Cultivar Discrimination of Korean and Chinese Boxthorn (Lycium chinense Mill. and Lycium barbarum L.) using SSR Markers (SSR 마커를 이용한 한국산과 중국산 구기자의 품종 판별)

  • Chung, Jong-Wook;Lee, Gi-An;Lee, Sok-Su;Bang, Kyong-Hwan;Park, Chung-Berm;Park, Yong-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.6
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    • pp.445-451
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    • 2009
  • This study was undertaken to develop a technique of discrimination using SSR makers in boxthorn cultivars. Forty one boxthorn cultivars, which were collected from Korea and China, were evaluated by 10 SSR markers. Total of 61 alleles were detected, ranging from 3 to 13 with an average of 6.1 alleles per locus. The averages of gene diversity and PIC values were 0.482 and 0.428, with a range from 0.25 (GB-LCM-022 and GB-LCM-087) to 0.83 (GB-LCM-167) and from 0.24 (GB-LCM-022 and GB-LCM-087) to 0.81 (GB-LCM-167), respectively. Five markers out of 10 markers, GB-LCM-022, GB-LCM-075, GB-LCM-104, GB-LCM-167 and GB-LCM-217, were selected as key markers for discrimination in boxthorn cultivars. All of boxthorn cultivars were individually distinguished by the combination of five SSR markers.

Characterization of Protocatechuate 4,5-Dioxygenase Induced from p-Hydroxybenzoate -Cultured Pseudomonas sp. K82

  • Yun, Sung-Ho;Yun, Chi-Young;Kim, Seung-Il
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.152-155
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    • 2004
  • Pseudomonas sp. K82 has been reported to be an aniline-assimilating soil bacterium. However, this strain can use not only aniline as a sole carbon and energy source, but can also utilize benzoate, p-hydroxybenzoate, and aniline analogues. The strain accomplishes this metabolic diversity by using dif-ferent aerobic pathways. Pseudomonas sp. K82, when cultured in p-hydroxybenzoate, showed extradiol cleavage activity of protocatechuate. In accordance with those findings, our study attempted the puri-fication of protocatechuate 4,5-dioxygenase (PCD 4,5). However the purified PCD 4,5 was found to be very unstable during purification. After Q-sepharose chromatography was performed, the crude enzyme activity was augmented by a factor of approximately 4.7. From the Q-sepharose fraction which exhibited PCD 4,5 activity, two subunits of PCD4,5 (${\alpha}$ subunit and ${\beta}$ subunit) were identified using the N-terminal amino acid sequences of 15 amino acid residues. These subunits were found to have more than 90% sequence homology with PmdA and PmdB of Comamonas testosteroni. The molecular weight of the native enzyme was estimated to be approximately 54 kDa, suggesting that PCD4,5 exists as a het-erodimer (${\alpha}$$_1$${\beta}$$_1$). PCD 4,5 exhibits stringent substrate specificity for protocatechuate and its optimal activity occurs at pH 9 and 15 $^{\circ}C$. PCR amplification of these two subunits of PCD4,5 revealed that the ${\alpha}$ subunit and ${\beta}$ subunit occurred in tandem. Our results suggest that Pseudomonas sp. K82 induced PCD 4,5 for the purpose of p-hydroxybenzoate degradation.

A Study on the Community Structure of Vegetation Landscape in Naejangsan National(I) (내장산국립공원 식물경관의 군집구조에 관한 연구(I))

  • 이규완;심경구
    • Journal of the Korean Institute of Landscape Architecture
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    • v.21 no.2
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    • pp.50-67
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    • 1993
  • This study analyzed the distribution and structure of the vegetation landscape in Naejangsan National Park. The plant distribution was investigated using a field survey. There were 72 sites sampled using the clumpled sampling method. The data derived from the investigation was analyzed using the quantitative analysis of Bray-Curtis method. The analysis was performed by the classification of TWINSPAN. The ordination of DCA and RA was used for the species composition and successional trends. The results are as follows; 1. Quercus. serrata-Q. variabilis community as 21.76(16.49$\textrm{km}^2$), was the largest community in the actual vegetation of the Naejangsan National Park. The degree of green naturality index 8 area covered 64.8% of the study area and the index 6 area did 16%. 2. Classified by the ordination of DCA and environmental variables, such as the plant community was divided into seven groups according to the altitude and forming groups; Chamaecy Paris. pisifera-P.densiflora community, P.densiflora community, Q.variailis community, T.nuciofera-A.palmatum community. 3. Ther species diversity and evenness indices of C.pisifera-P.densiflora community, P.densiflora community appeared low but C.laxiflora communitywhich was the most stable community in the study area. 4. The similarity indices between Q.variabilis community and Q.serrata-Q.variabilis community were calculated 58.84%, but those between other communities were comparatively low level. 5. The successional trends of DBH class seem to be from C. pisifera-P.densiflora community, P.densiflora community to Quercus species community and from Q.variabilis community, Q.serrata-Q.variabilis community to Carpinus species community. 6. Results suggested that the successional trends in Naejangsan National Park; P.densiflora community\longrightarrowQ.variablilis community, Q.serrata-Q.variabilis community\longrightarrowC.laxiflora community in sequence.

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A report of 17 unrecorded bacterial species of Korea belonging to the phylum Bacteroidetes

  • Cho, Jang-Cheon;Seong, Chi Nam;Joh, Kiseong;Cha, Chang-Jun;Bae, Jin-Woo;Yi, Hana;Lee, Soon Dong;Kim, Myung Kyum;Yoon, Jung-Hoon
    • Journal of Species Research
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    • v.7 no.3
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    • pp.231-239
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    • 2018
  • While screening indigenous prokaryotic species in Republic of Korea in 2017, a total of 17 bacterial strains assigned to the phylum Bacteroidetes were isolated from a variety of environmental habitats including water of fountain, tidal flat, plant root, soil, the gut of Russian grayling butterfly, ginseng field, seawater, lagoon and seashore sand. From the 16S rRNA gene sequence similarity of more than 98.7% and the formation of a robust phylogenetic clade with the closest species, it was found that the 17 strains belong to independent and recognized bacterial species. There has been no official report that the identified 17 species have been previously isolated in the Republic of Korea. Thus, 15 species in 10 genera of one family in the order Flavobacteriales, one species in one genus of one family in the order Cytophagales, and one species in one genus of one family in the order Sphingobacteriales are proposed as unrecorded species of the phylum Bacteroidetes found in the Republic of Korea. Gram reaction, colony and cell morphology, basic phenotypic characteristics, isolation source, taxonomic status, strain ID and other information are described in the species descriptions.