• Title/Summary/Keyword: phylogenetic study

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Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.

Biochemical and cultural characteristics of mineral-solubilizing Acinetobacter sp. DDP346 (미네랄 가용화능을 갖는 Acinetobacter sp. DDP346의 생화학적 및 배양학적 특성)

  • Kim, Hee Sook;Lee, Song Min;Oh, Ka-Yoon;Kim, Ji-Youn;Lee, Kwang Hui;Lee, Sang-Hyeon;Jang, Jeong Su
    • Journal of Applied Biological Chemistry
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    • v.64 no.4
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    • pp.333-341
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    • 2021
  • In this study, to select strains suitable as microbial agent from among rhizosphere microorganisms present in rhizosphere soil and roots, the mineral solubilization ability, antifungal activity against 10 types of plant pathogenic fungi, and plant growth-promoting activity of rhizosphere microorganisms were evaluated. As a result, DDP346 was selected because it has solubilization ability of phosphoric acid, calcium carbonate, silicon, and zinc; nitrogen fixing ability; production ability of siderophore, indole-3-acetic acid, and aminocyclopropane-1-carboxylate deaminase; and antifungal activity against seven types of plant pathogenic fungi. DDP346 showed a 99.9% homology with Acinetobacter pittii DSM 21653 (NR_117621.1); phylogenetic analysis also revealed a close relationship with Acinetobacter pittii based on the 16S rRNA base sequence. The growth conditions of DDP346 were identified as temperatures in the range of 10-40 ℃, pH in the range of 5-11, and salt concentrations in the range of 0-5%. In addition, a negative correlation coefficient (r2 = -0.913, p <0.01) was shown between pH change and the solubilized phosphoric acid content of Acinetobacter sp. DDP346, and this is assumed to be due to the organic acid generated during culture. Consequently, through the evaluation of its mineral solubilization ability, antifungal activity against plant pathogenic fungi, and plant growth-promoting activity, the potential for the utilization of Acinetobacter sp. DDP346 as a multi-purpose microbial agent is presented.

Occurrence of a Natural Intergeneric Hybrid between a Female Tanakia lanceolata and a Male Rhodeus pseudosericeus (Cypriniformes: Cyprinidae) in Daecheoncheon Stream Flowing into the Yellow Sea in the Republic of Korea (서해안 독립 하천 대천천에서 납자루 Tanakia lanceolata (♀)와 한강납줄개 Rhodeus pseudosericeus(♂)의 자연 속간잡종 출현)

  • Kim, Yong Hwi;Sung, Mu Sung;Yun, Bong Han;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.33 no.2
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    • pp.45-56
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    • 2021
  • A male, presumed to be an intergeneric hybrid between Tanakia lanceolata and Rhodeus pseudosericeus, was collected in the Boryeong Daecheoncheon Stream flowing into the Yellow Sea in the Republic of Korea. Morphological and molecular phylogenetic analyses were performed to discriminate the definite origin of the estimated natural hybrid. As a result of the morphological analysis, the color of the dorsal and anal fin rays edges of the natural hybrid individual, the upper and lower body colors followed the morphological characteristics of T. lanceolata, and that blue longitudinal stripe in the center of the caudal peduncle, the incomplete lateral line, and the barbels absent followed the morphological characteristics of R. pseudosericeus. In addition, as a result of the cytochrome b (cytb) gene analysis of mitochondrial DNA (mtDNA), the natural hybrid showed a nucleotide sequence similarity of 99.82 to 100% with T. lanceolata, and the maternal species was identified as T. lanceolata. As a result of the recombination activating gene 1 (rag1) gene analysis of nuclear DNA (nDNA), the natural hybrid showed double peaks pattern reflecting both the single nucleotide polymorphism sites (38 bp) between T. lanceolata and R. pseudosericeus, and the paternal species was identified as R. pseudosericeus. Therefore, a natural hybrid estimated male of Acheilognathinae analyzed in this study was found to be an intergeneric hybrid between a female T. lanceolata and a male R. pseudosericeus.

Generation of Bacterial Blight Resistance Rice with Transcription Factor OsNAC69-overexpressing (전사인자 OsNAC69-과발현을 통한 흰잎마름병 저항성 벼 제작)

  • Park, Sang Ryeol;Cha, Eun-Mi;Moon, Seok Jun;Shin, Dongjin;Hwang, Duk-Ju;Ahn, Il-Pyung;Bae, Shin-Chul
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.457-463
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    • 2011
  • Plant specific gene family, NAC (NAM, ATAF, and CUC) transcription factors have been characterized for their roles in plant growth, development, and stress tolerance. In this study, we isolated OsNAC69 gene and analyzed expression level by inoculation of bacterial leaf blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo). NAC transcription factor family can be divided into five groups (I-V). On the basis of phylogenetic analysis, OsNAC69 was fall into group II. OsNAC69 was strongly induced 1 hr after infected with Xoo. To investigate its biological function in the rice, we constructed vector for overexpression in rice, and then generated transgenic rice lines. Gene expression of OsNAC69-overexpressed transgenic rice lines were analyzed by northern blot. Analysis of disease resistance to pathogen Xoo, nine OsNAC69-overexpressed transgenic rice lines showing high expression level of OsNAC69 were shown more resistant than wild type. These results suggest that OsNAC69 gene may play regulatory role during pathogen infection.

Exocyclic GpC DNA methyltransferase from Celeribacter marinus IMCC12053 (Celeribacter marinus IMCC12053의 외향고리 GpC DNA 메틸트랜스퍼라아제)

  • Kim, Junghee;Oh, Hyun-Myung
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.103-111
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    • 2019
  • DNA methylation is involved in diverse processes in bacteria, including maintenance of genome integrity and regulation of gene expression. CcrM, the DNA methyltransferase conserved in Alphaproteobacterial species, carries out $N^6$-adenine or $N^4$-cytosine methyltransferase activities using S-adenosyl methionine as a co-substrate. Celeribacter marinus IMCC12053 from the Alphaproteobacterial group was isolated from a marine environment. Single molecule real-time sequencing method (SMRT) was used to detect the methylation patterns of C. marinus IMCC12053. Gibbs motif sampler program was used to observe the conversion of adenosine of 5'-GANTC-3' to $N^6$-methyladenosine and conversion of $N^4$-cytosine of 5'-GpC-3' to $N^4$-methylcytosine. Exocyclic DNA methyltransferase from the genome of strain IMCC12053 was chosen using phylogenetic analysis and $N^4$-cytosine methyltransferase was cloned. IPTG inducer was used to confirm the methylation activity of DNA methylase, and cloned into a pQE30 vector using dam-/dcm- E. coli as the expression host. The genomic DNA and the plasmid carrying methylase-encoding sequences were extracted and cleaved with restriction enzymes that were sensitive to methylation, to confirm the methylation activity. These methylases protected the restriction enzyme site once IPTG-induced methylases methylated the chromosome and plasmid, harboring the DNA methylase. In this study, cloned exocyclic DNA methylases were investigated for potential use as a novel type of GpC methylase for molecular biology and epigenetics.

Selection of Entomopathogenic Fungus Isaria javanica FT333 for Dual Control of Thrips and Anthracnose (총채벌레 및 고추탄저병의 동시 방제를 위한 곤충병원성 곰팡이 Isaria javanica FT333 선발)

  • Lee, Moran;Jeong, Hyeju;Kim, Jaeyoon;Kim, Dayeon;Ahn, Seung Ho;Lee, SangYeob;Han, Ji Hee
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.479-490
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    • 2018
  • Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.

Geographic variation of Grey-capped Greenfinch (Chloris sinica) in Korea (한국에서 방울새(Grey-capped Greenfinch, Chloris sinica)의 지리적 변이에 대한 연구)

  • Park, Jong-Gil;Kim, Joo-Eun;Jin, Kyoung-Soon;Park, Chungoo;Nam, Dong-Ha
    • Korean Journal of Ornithology
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    • v.25 no.2
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    • pp.117-125
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    • 2018
  • The Grey-capped greenfinch (Chloris sinica) is a polytypic species that exhibits complicated geographical variation in morphology. This study provides an intraspecific phylogeographic variability of C. sinica populations in Korea with their morphometric data. The observed morphometric variations were that Ulleung island population was morphologically distinct in bill length and depths as compared to the mainland populations. Phylogenetic relationships among mitochondrial COX1 regions provided evidence for genetic differentiation between Ulleung and mainland populations. However, their genetic distances and nucleotide diversities were very low, highlighting their recent divergence. The needs for additional research is heightened to substantiate if the genetic clines in different localities may arise in C. sinica subspecies, each of which could have different breeding and wintering habitats, distribution patterns, and migration pathways.

Characterization of Miniimonas sp. S16 isolated from activated sludge (활성슬러지로부터 분리된 Miniimons sp. S16 세균의 특성)

  • Koh, Hyeon-Woo;Kim, Hongik;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.242-247
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    • 2019
  • Biological factors (e.g. microorganism activity) in wastewater treatment plant (WWTP) play essential roles for degradation and/or removal of organic matters. In this study, to understand the microbial functional roles in WWTP, we tried to isolate and characterize a bacterial strain from activated sludge sample. Strain S16 was isolated from the activated sludge of a municipal WWTP in Daejeon metropolitan city, the Republic of Korea. The cells were a Gram-stain-positive, non-motile, facultative anaerobe, and rod-shaped. Strain S16 grew at a temperature of $15{\sim}40^{\circ}C$ (optimum, $30^{\circ}C$), with 0~9.0% (w/v) NaCl (optimum, 1.0~2.0%), and at pH 5.5~9.0 (optimum, pH 7.0~7.5). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S16 was most closely related to the unique species Miniimonas arenae NBRC $106267^T$ (99.79%, 16S rRNA gene sequence similarity) of the genus Miniimonas. The cell wall contained alanine, glutamic acid, serine, and ornithine. Although the isolation source of the type strain NBRC $106267^T$ which considered as a marine microorganism is sea sand, that of strain S16 is terrestrial environment. It might raise an ecological question for habitat transition. Therefore, comparative genome analysis will be valuable investigation for shedding light on their potential metabolic traits and genomic streamlining.

Development of a bioassay for screening of resistance to Tomato spotted wilt virus isolate from Korea (국내 분리 토마토반점위조바이러스의 저항성 판별을 위한 생물검정법 개발)

  • Kwak, Hae-Ryun;Choi, Hyeon-Yong;Hong, Su-Bin;Hur, On-Sook;Byun, Hee-Seong;Choi, Hong-Soo;Kim, Mikyeong
    • Korean Journal of Environmental Biology
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    • v.39 no.3
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    • pp.319-328
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    • 2021
  • Tomato spotted wilt virus (TSWV) is one of the most destructive viruses worldwide, which causes severe damage to economically important crops, such as pepper and tomato. In this study, we examined the molecular and biological characterization of a TSWV isolate (SW-TO2) infecting tomato and compared it to the recently reported isolates from boxthorn, butterbur, and angelica plants. The phylogenetic analysis based on the complete genome sequences confirmed that SW-TO2 was clustered with those of isolates from boxthorn and pepper in Korea with the maximum nucleotide identities ranging from 98% to 99%. We developed the bioassay method for screening TSWV resistance and tested some commercial pepper and tomato cultivars for resistance evaluation of four isolates of TSWV. TSWV resistance was evaluated as TSWV resistance when all the following three conditions were satisfied: first, when symptoms of necrotic spots or no symptoms were present in the inoculated leaves; second, when there were no symptoms in the upper leaves; and third, when the upper leaves were negative as a result of RT-PCR diagnosis.

Transcriptional regulation of chicken leukocyte cell-derived chemotaxin 2 in response to toll-like receptor 3 stimulation

  • Lee, Seokhyun;Lee, Ra Ham;Kim, Sung-Jo;Lee, Hak-Kyo;Na, Chong-Sam;Song, Ki-Duk
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1942-1949
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    • 2019
  • Objective: Leukocyte cell-derived chemotaxin 2 (LECT2) is associated with several physiological processes including inflammation, tumorigenesis, and natural killer T cell generation. Chicken LECT2 (chLECT2) gene was originally identified as one of the differentially expressed genes in chicken kidney tissue, where the chickens were fed with different calcium doses. In this study, the molecular characteristics and gene expression of chLECT2 were analyzed under the stimulation of toll-like receptor 3 (TLR3) ligand to understand the involvement of chLECT2 expression in chicken metabolic disorders. Methods: Amino acid sequence of LECT2 proteins from various species including fowl, fish, and mammal were retrieved from the Ensembl database and subjected to Insilco analyses. In addition, the time- and dose-dependent expression of chLECT2 was examined in DF-1 cells which were stimulated with polyinosinic:polycytidylic acid (poly [I:C]), a TLR3 ligand. Further, to explore the transcription factors required for the transcription of chLECT2, DF-1 cells were treated with poly (I:C) in the presence or absence of the nuclear factor ${\kappa}B$ ($NF{\kappa}B$) and activated protein 1 (AP-1) inhibitors. Results: The amino acid sequence prediction of chLECT2 protein revealed that along with duck LECT2 (duLECT2), it has unique signal peptide different from other vertebrate orthologs, and only chLECT2 and duLECT2 have an additional 157 and 161 amino acids on their carboxyl terminus, respectively. Phylogenetic analysis suggested that chLECT2 is evolved from a common ancestor along with the actinopterygii hence, more closely related than to the mammals. Our quantitative polymerase chain reaction results showed that, the expression of chLECT2 was up-regulated significantly in DF-1 cells under the stimulation of poly (I:C) (p<0.05). However, in the presence of $NF{\kappa}B$ or AP-1 inhibitors, the expression of chLECT2 is suppressed suggesting that both $NF{\kappa}B$ and AP-1 transcription factors are required for the induction of chLECT2 expression. Conclusion: The present results suggest that chLECT2 gene might be a target gene of TLR3 signaling. For the future, the expression pattern or molecular mechanism of chLECT2 under stimulation of other innate immune receptors shall be studied. The protein function of chLECT2 will be more clearly understood if further investigation about the mechanism of LECT2 in TLR pathways is conducted.