• Title/Summary/Keyword: Phylogenetic

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Analysis of Species Variety and Physiological Characteristics of Denitrifying Oligotrophic Bacteria Isolated from the Specific Environment in Korea (국내 특수 생태환경의 탈질 저영양 세균의 종 다양성 및 생리적 특성 분석)

  • Lee, Chang-Muk;Weon, Hang-Yeon;Kwon, Soon-Wo;Kang, Han-Chul;Koo, Bon-Sung;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.210-217
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    • 2011
  • In an effort to isolate novel bacteria for the bioremediation of over-fertilized soils, we identified 135 denitrifying cells out of 3,471 oligotrophic bacteria pools (3.9%) using a denitrification medium supplemented with potassium nitrate as the sole nitrogen source. Soil samples were taken from ecologically well-conserved areas, including a mountain swamp around the demilitarized zone (Yongneup), two ecoparks (Upo and the Mujechi bog), and ten representative islands around the Korean peninsula (Jejudo, Daecheongdo, Socheongdo, Baekryeongdo, Ulrungdo, Dokdo, Geomundo, Hongdo, Huksando and Yeonpyeongdo). All of the 135 bacteria produced nitrogen gas from the denitrification medium, and were proved to be nitrate reductase positive by API-BioLog tests. Phylogenetic analysis using 16S rDNA sequences revealed that the 135 bacteria consisted of 44 different genera. Along with the most prominent, Proteobacteria (87.4%), we identified denitrifying bacteria from Firmicutes (9.4%), Actinobacteria (2.4%), and Bacteroidetes (0.8%). Physiological analyses of the 44 representative denitrifying bacteria, under various pH levels, growth temperatures and salt stresses, revealed 12 favorable denitrifying strains for soil bioremediation.

Biochemical Characterization of Recombinant L-Asparaginase (AnsA) from Rhizobium etli, a Member of an Increasing Rhizobial-Type Family of L-Asparaginases

  • Moreno-Enriquez, Angelica;Evangelista-Martinez, Zahaed;Gonzalez-Mondragon, Edith G.;Calderon-Flores, Arturo;Arreguin, Roberto;Perez-Rueda, Ernesto;Huerta-Saquero, Alejandro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.292-300
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    • 2012
  • We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

Salinity Stress Resistance Offered by Endophytic Fungal Interaction Between Penicillium minioluteum LHL09 and Glycine max. L

  • Khan, Abdul Latif;Hamayun, Muhammad;Ahmad, Nadeem;Hussain, Javid;Kang, Sang-Mo;Kim, Yoon-Ha;Adnan, Muhammad;Tang, Dong-Sheng;Waqas, Muhammad;Radhakrishnan, Ramalingam;Hwang, Young-Hyun;Lee, In-Jung
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.893-902
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    • 2011
  • Endophytic fungi are little known for their role in gibberellins (GAs) synthesis and abiotic stress resistance in crop plants. We isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (CF) on the GAs biosynthesis mutant rice line - Waito-C. CF bioassay showed that endophyte GMH-1B significantly promoted the growth of Waito-C compared with controls. GMH-1B was identified as Penicillium minioluteum LHL09 on the basis of ITS regions rDNA sequence homology and phylogenetic analyses. GC/MS-SIM analysis of CF of P. minioluteum revealed the presence of bioactive $GA_4$ and $GA_7$. In endophyte-soybean plant interaction, P. minioluteum association significantly promoted growth characteristics (shoot length, shoot fresh and dry biomasses, chlorophyll content, and leaf area) and nitrogen assimilation, with and without sodium chloride (NaCl)-induced salinity (70 and 140 mM) stress, as compared with control. Field-emission scanning electron microcopy showed active colonization of endophyte with host plants before and after stress treatments. In response to salinity stress, low endogenous abscisic acid and high salicylic acid accumulation in endophyte-associated plants elucidated the stress mitigation by P. minioluteum. The endophytic fungal symbiosis of P. minioluteum also increased the daidzein and genistein contents in the soybean as compared with control plants, under salt stress. Thus, P. minioluteum ameliorated the adverse effects of abiotic salinity stress and rescued soybean plant growth by influencing biosynthesis of the plant's hormones and flavonoids.

Phylogeny of the Polygonatum odoratum Complex Inferred from Multiple cpDNA and Nuclear RNApol2_i23 Sequence Data (Ruscaceae) (엽록체 DNA 및 핵 DNA RNApol2_i23에 근거한 둥굴레복합체 (Ruscaceae)의 계통 연구)

  • Park, Jeong-Mi;Chung, Kyong-Sook;Oh, Byoung-Un;Jang, Chang-gee
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.353-360
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    • 2011
  • The sequence data of the plastid DNA (trnL-F IGS, trnL intron, and trnH-psbA) and nuclear DNA (RNApol2_i23) markers were utilized to study phylogenetic relationships among the taxa in the Polygonatum odoratum complex (Ruscaceae). European P. odoratum individuals form a clade with a high bootstrap value, which is a sister to the clade of Korean P. odoratum var. odoratum, P. odoratum var. pluriflorum and P. robustum. The formation of the clade with P. odoratum var. odoratum, P. robustum, and one accession of P. odoratum var. pluriflorum indicates geological speciation in isolated populations in the islands following dispersal events from the mainlands. All data sets form two major clades, which are congruent with the subgroups divided by the basic chromosome numbers (x = 9 and x = 10). Although it is not easy to test the hypothesis of the decrease in the basic chromosome number due to scatter taxon sampling in this study, the molecular data strongly suggested that aneuploidy plays an important role in lineage diversification in the genus Polygonatum. The cytological data was not strongly supported by the cpDNA sequences. Further investigations of the cytological, morphological, and geographical characteristics with comprehensive sampling are desired to understand the evolution and lineage diversification in the genus.

Molecular Identification and Distribution Pattern of Fish Eggs Collected around Jejudo Island (제주 연안해역 어란의 분자동정과 분포양상)

  • Han, Song-Hun;Kim, Maeng Jin;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.284-292
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    • 2015
  • Monthly variation of species composition and abundance of fish eggs were examined to know the spawning time and location of the fishes inhabiting the coastal region of Jejudo Island. Samplings had been performed at the four locations (Jeju port, Seongsanpo, Seogwipo port and Chagwido) with a bongo net which was towed monthly at the sea surface from August 2006 to July 2007. The fish eggs were identified based on phylogenetic analyses with the DNA sequences generated through PCR-amplification and sequencing of the mitochondrial cytochrome b gene. Among a total of 43 taxa classified during the study period, 34 were identified to species, 4 to families, and the remaining 5 were unidentified. Of them, 23 taxa were occurred at Jeju port, 21 at Seongsanpo, 19 at Seogwipo port and 18 at Chagwido, whereas 15 taxa were found in September 2006, 12 in June 2007, 6 to 8 in August 2006 and January~May 2007, 5 in each October and November 2006, 3 in each December 2006 and July 2007. Among 34 species of fish eggs, Engraulis japonicus and Callanthias japonicus most frequently appeared at 16 times out of 48 observations in total. When those eggs were collected, the surface seawater temperature ranged $14.0{\sim}28.6^{\circ}C$ for E. japonicus and $14.9{\sim}20.5^{\circ}C$ for C. japonicus. The success rates of PCR-amplification and species identification in this study were 68.3% and 79.1%, respectively.

Isolation and Characterization of Comamonase sp. and Microbacterium sp. from Deep Blue Sediment Dye of Polygoum tinctoria, Niram (쪽 염료 니람으로부터 Comamonas sp.와 Microbacterium sp.의 분리 및 특성분석)

  • Jang, Seong Eun;Lee, Nam Keun;Lee, Yuri;Choi, Mee-Sung;Jeong, Yong-Seob
    • KSBB Journal
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    • v.28 no.1
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    • pp.60-64
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    • 2013
  • Two strains were isolated from the traditional Deep Blue Sediment Dye of Polygoum tinctoria, Niram, and temporarily named Niram A and Niram B, respectively. The phylogenetic analysis revealed that strain Niram A and B were closely related to the members of the genus Comamonas and Microbacterium, respectively. Strain Niram A exhibited the highest 16S rRNA gene sequence similarity to C. aquatica LMG $2370^T$ (98.06%). Strain Niram B showed 100% homology with M. oxydans DSM 20578T and M. maritypicum DSM $12512^T$. The growth of the strain Niram A and B was not inhibited in Niram medium containing high calcium concentration without free sugar as carbon source. The reducing Niram is greenish. Therefore, the reducing ability on the Niram of the strains Niram A and B were determined with the color difference of the $a^*$ values of Niram fermented-fluids. The $a^*$ value indicates the level of redness (positive value) or greenness (negative value). The green color is increasing towards the negative value. In all samples fermented for 10 days, the $a^*$ values among samples were no significant difference. However, samples fermented for 15 days have an appreciable change. After fermentation for 15 days, the control Niram sample had $-3.96{\pm}0.02$ of the $a^*$ value. On the other hand, the Niram samples fermented with the strain Niram A and B showed $-4.20{\pm}0.02$ of the $a^*$ value and $-7.86{\pm}0.03$ of the $a^*$ value, respectively. In the reducing ability on the Niram, the strain Niram B was significantly better than the strain Niram A.

Phylogenic Analysis of Locusta migratoria (Orthoptera: Acridae) in Haenam-gun and Muan-gun, Jeollanam-do, Korea Using Mitochondrial NADH dehydrogenase subunits (전라남도 해남과 무안의 풀무치 개체군에 대한 마이토콘드리아 NADH dehydrogenase subunit 들을 이용한 계통분석)

  • Lee, Gwan Seok;Kim, Young Ha;Jung, Jin Kyo;Koh, Young Ho
    • Korean journal of applied entomology
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    • v.56 no.4
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    • pp.371-376
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    • 2017
  • In a nationwide survey of the occurrence and density of the migratory locust (Locusta migratoria), high density was continuously observed in the reclaimed areas of Mangun-myeon in Muan-gun, Jeollanam-do, and Sanye-myeon in Haenam-gun, Jeollanam-do, Korea. We have analyzed the nucleotide sequences of NADH dehydrogenase subunit (NAD) 2, NAD4, and NAD5 genes in order to determine the origins of the migratory locusts at two sites. According to the analysis, the migratory locusts in Haenam-gun were closely related with those in Liaoning Province and Heilongjiang Province in the northeast China. In contrast, the migratory locusts in Muan-gun were most similar to those in Japan. Because Korean migratory locusts were not included in the previous global study on the evolution and migration of migratory locusts, we did not know the origin of Korean migratory locusts, earlier. Phylogenetic analyses this study suggested that the migratory locusts from the northeast Chinese population might have migrated and settled in Haenam-gun in Korea. Moreover, another northeast Chinese population might have migrated to Muan-gun in Korea though Sakhalin, Russia and Hokkaido, Japan. However, the possibility that the migratory locusts moved from northeast China might be isolated from each other in Korea, and that the Muan population might migrate to Japan cannot be excluded.

First Report of Asplenium yoshinagae (Aspleniaceae) from Korea and Its Phylogenetic Position Based on Morphology (미기록 가거꼬리고사리(꼬리고사리과)의 계통학적 위치)

  • Lee, Chang Shook;Kim, Jin Ohk;Lee, Nam Sook
    • Korean Journal of Plant Taxonomy
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    • v.38 no.2
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    • pp.79-91
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    • 2008
  • Asplenium yoshinagae (Aspleniaceae), previously known only from Japan, southwest China to Himalaya, was found in Gageo-do, Heuksan-myeon, Sinan-gun, Jeollanam-do. This species is similar to A. trichomonas, A. tripteropus, A. boreale, A. normale and A. oligophlebium by having gemmae and auricle of pinna, and distinguished from the latters by distinct stipe length, stalk of pinna, acute apex of pinna, length of indusium and shape of sorus. The Local name, Ga-geo-kko-ri-go-sa-ri, was newly given considering the locality. To reveal the interspecific relationships within the genus Asplenium in Korea, cladistic analysis was performed for 22 taxa of Asplenium as ingroup and 2 taxa of Diplazium as outgroup from Korea based on 20 morphological characters. As the results, the genus Asplenium seperated strongly from outgroup, and divided into 4 clades. Asplenium yoshinagae belong to the third clade. A. hondoense N. Murtakami & S. I. Hatanaka, which contained in the second clade, had treated as Hymenasplenium, but this results supported that this taxon may be contained in Asplenium, and also, Asplenium ruprechtii, not in Comptosorus. The morphological characters and illustrations of the species are provided together with photographs of habitat.

Genetic differences between Korean-Japanese and Chinese-Taiwanese Dendrobium moniliforme (L.) Sw. (한국-일본과 중국-대만 석곡의 유전적 차이)

  • Kim, Young-Kee;Kang, Kyung-Won;Kim, Ki-Joong
    • Korean Journal of Plant Taxonomy
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    • v.45 no.2
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    • pp.145-157
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    • 2015
  • The nucleotide sequences of nuclear ribosomal ITS regions and chloroplast rbcL, matK and psbA-trnH regions of 30 individuals of Dendrobium moniliforme from several localities in four countries and 28 related species of Dendrobium were compared to investigate the genetic differences among Korean, Japanese, Taiwanese and Chinese D. moniliforme, and to verify the homogeneity of D. moniliforme, which is used as a traditional medicine in East Asia. A phylogenetic analysis showed that Korean D. moniliforme and Japanese D. moniliforme form a monophyletic group, with no significant differences between their nucleotide sequences. This confirms that they are the same species. However, the Chinese and Taiwanese D. moniliforme were polyphyletic. Various species related to D. moniliforme were located between the Korean-Japanese D. moniliforme and the Chinese-Taiwanese D. moniliforme, and other related species were found between individuals of Chinese-Taiwanese D. moniliforme. D. moniliforme is described in Japan, providing evidence that the Korean-Japanese D. moniliforme is the original species. In addition, our data suggest that the Chinese-Taiwanese D. moniliforme complex is a mixture of a range of other species. Further studies are required to understand the taxonomic identity of this species. In the Korean-Japanese D. moniliforme, there were almost no genetic differences among the localities, whereas the genetic heterogeneity was high among individuals of the Chinese-Taiwanese D. moniliforme.

Characteristics of Biocellulose by Gluconobacter uchimurae GYS15 (Gluconobacter uchimurae GYS15 균주로부터 생산되는 Biocellulose의 특성 확인)

  • Lee, Young Sun;Kim, Jae Young;Cha, Mi Yeon;Kang, Hee Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.247-255
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    • 2016
  • In order to select a strain that forms a Biocellulose (BC), strain producing acetic acid was selected from commercially available kombucha. Through SM broth it was confirmed that the strain is a gram negative bacteria in the form of rods having no motility through a phase contrast microscope. The result of phylogenetic inference analysis based on 16S rDNA sequence analysis for the identification of strains was most closely related to Gluconobacter uchimurae (G. uchimurae) and was named G. uchimurae GYS15 strain. The strain showed the highest degree of growth when cultured for 14 days under the conditions of pH 5 and $25^{\circ}C$. Moreover, it showed the highest degree of growth in a Glucose addition disaccharide as the optimum carbon source sucrose and fructose. Also, 0.5% NaCl, upon the addition of Malto extract, showed the highest degree of growth. Based on investigation by the optimum growth conditions to confirm the physical properties of BC obtained by culturing G. uchimurae GYS15 strains. The surface structure was observed through an scanning electron microscope (SEM) showed a high networks structure. It until $8.6{\pm}0.38$ times when the water holding capacity is re-absorbed and re-absorbed holding oil up to $6.6{\pm}0.51$ times confirmed. In conclusion, using these material properties, it was possible to confirm the possibility of a variety of cosmetic materials and mask pack materials.