• Title/Summary/Keyword: ribosomal DNA.

Search Result 650, Processing Time 0.024 seconds

Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
    • /
    • v.9 no.2
    • /
    • pp.156-165
    • /
    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

  • PDF

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
    • /
    • v.42 no.4
    • /
    • pp.317-323
    • /
    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

Isolation and Identification of a Streptomyces sp. that Produces Antibiotics Against Multidrug - Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii의 생장을 억제하는 항생물질을 생산하는 방선균의 분리.동정 및 항균효과)

  • Rhee, Ki-Hyeong
    • Microbiology and Biotechnology Letters
    • /
    • v.39 no.1
    • /
    • pp.37-42
    • /
    • 2011
  • I isolated the actinomycete strain KH223 from soil samples collected from the Kye Ryong mountain area. This strain is antagonistic to the multidrug-resistant Acinetobacter baumannii. KH223 was confirmed as belonging to the genus Streptomyces based on the scanning electronmicroscopy(SEM) observations of the diaminopimelicacid(DAP) type and morphological and physiological characteristics. Comparison of the 16S rDNA nucleotide sequences revealed that KH223 has a relationship with Streptomyces galbus. Production of antibiotics by KH223 was most favorable when cultured on a glucose, polypeptone, and yeast extract(PY) medium for 6 days at 27$^{\circ}C$. The supernatant was found to exhibit an antimicrobial effect on various kinds of bacteria and fungi. Particularly, butanol and ethylacetate extracts of KH223 and cyclo(trp-trp) exhibited significant activity against A. baumannii at concentration ranges of 0.8-12.5 ${\mu}g$/mL, 5.0-25 ${\mu}g$/mL and 12.5${\rightarrow}$100 ${\mu}g$/mL, respectively. Moreover, in contrast to cyclo(trp-trp) had shown to activity against Micrococcus luteus JCM 1464 at the concentration of 12.5 ${\mu}g$/mL, the butanol extract of KH223 showed significant activity against Bacillus subtilis IAM 1069 and Micrococcus luteus JCM 1464 at the concentration of 0.4 and 0.8 ${\mu}g$/mL, respectively. These results suggest that KH223 may have a great potential in the production of new antibiotics to combat multidrug-resistant pathogens and further studies may be warranted for the same.

Probiotic Potential of Lactobacillus Isolates (Lactobacillus 분리균주의 프로바이오틱스로서의 가능성 검토)

  • Bang, Ji-Hun;Shin, Hwa-Jin;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
    • /
    • v.22 no.2
    • /
    • pp.251-258
    • /
    • 2012
  • The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from animal feces. BCNU 9041 and BCNU 9042 isolates were assigned to Lactobacillus brevis on the basis of their physiological properties and 16S ribosomal DNA sequences analysis. They were confirmed as safe bioresources because of their non-hemolytic activities and non-production of harmful ${\beta}$-glucosidase, ${\beta}$-glucuronidase, tryptophanase, or urease. These isolates were also highly resistant to acid (at pH 2.5) and bile acids (at concentration of 0.3%, 0.6%, and 1% oxgall). In addition, they exhibited good antibacterial activity against food-borne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Shigella sonnei. Furthermore, it was demonstrated that they have the highest levels of hydrophobicity and that they showed bile salt hydrolytic and cholesterol assimilation activity. These results suggest that BCNU 9041 and 9042 have good potential for application in functional foods and health-related products.

Characteristics of Brown Rot Caused by Monilinia fructicola on Stone Fruit in Korea (핵과류 잿빛무늬병을 일으키는 Monilinia fructicola 병해 특성)

  • Oh, Hun-Tak;Choi, In-Young;Kim, Ju;Na, Young-Eun;Lee, Wang-Hyu;Lee, Kui-Jae;Shin, Hyeon-Dong
    • Research in Plant Disease
    • /
    • v.23 no.4
    • /
    • pp.322-333
    • /
    • 2017
  • In June and July 2015 and 2017, typical signs and symptoms of brown rot were observed on the fruit of Japanese apricot, peach, apricot, Japanese plum, and sweet cherry with incidence levels of 2-5% in Jeonju and Imsil, Korea. Early symptoms were small, circular, light brown spots that eventually destroyed entire fruit. Small sporodochia later appeared on the surface. Conidia isolated from each host were one-celled, hyaline, lemon-shaped and borne in branched monilioid chains. The optimal temperature range for hyphal growth of all the isolates was $20-25^{\circ}C$. The growth of hyphae was faster on potato dextrose agar and oatmeal agar than others. Multiple alignments using the ITS sequences from different host showed that they matched each other (100%). The ITS sequences showed 100% identity to those of M. fructicola. Based on the morphological characteristics and phylogenetic analysis via internal transcribed spacer (ITS), all the isolate was identified as M. fructicola. Pathogenicity of representative isolates was proved by artificial inoculation, fulfilling Koch's postulates. This is the first confirmed report on brown rot caused by M. fructicola on stone fruit in Korea.

Dermatophytosis of the Four-toed Hedgehog Caused by Trichophyton erinacei

  • Yoon, Ji-Seon;Lee, Jong-Hyun;Yu, Do-Hyeon;Li, Ying-Hua;Lee, Mi-Jin;Iwasaki, T.;Park, Jin-Ho
    • Journal of Veterinary Clinics
    • /
    • v.25 no.3
    • /
    • pp.207-210
    • /
    • 2008
  • Trichophyton erinacei is a dermatophyte pathogen that infects both humans and hedgehogs. A two-month old female four-toed hedgehog presented to the Chonbuk Animal Medical Center with pruritus, excoriation and crust on her face for ten days. The owner of the hedgehog also exhibited the clinical signs of scaly erythema with fine vesicles on her neck. A presumptive diagnosis of dermatophytosis was made based on the results of an acetate tape preparation in which hyphae and chains of arthroconidia were observed. The crusts from the lesions were then cultured on Sabouraud Dextrose Agar for identification. After 10 days of incubation, downy colored colonies that had a central umbo with a white granular surface and a yellow pigment ring in the reverse were observed. Microscopic analysis revealed the presence of numerous teardrop shaped microconidia singly attached to the sides of the hyphae. In addition, 2-6 roomed macroconidia that were somewhat irregular in shape and size were present, and abundant intermediate sized spores were observed between the micro and macro conidia. To confirm that the culture was T. erinacei, the internal transcribed spacer region of the 5.8S phase of the ribosomal RNA gene (ITS1-5.8S-ITS2 rDNA) was amplified by PCR and then sequenced. A 679-base pair fragment of DNA was then compared with sequences in GenBank and found to be 99% homologous with sequences of T. erinacei (Z97997 and Z97996. The clinical signs were resolved after four weeks of treatment with oral and topical ketoconazole and chlorhexidine. To the best of our knowledge, this represents the first case of T. erinacei isolated from a four-toed hedgehog in Korea.

Chitinase Production and Isolation of Serratia plymuthica AL-1 Antagonistic to White Rot Fungi from Allium fistulosum Roots. (대파 뿌리로부터 흑색썩음균핵병균에 길항하는 Serratia plymuthica AL-1의 분리 및 Chitinase의 생산)

  • 주길재;이익희;김진호
    • Microbiology and Biotechnology Letters
    • /
    • v.30 no.2
    • /
    • pp.135-141
    • /
    • 2002
  • This study was carried out to isolate antagonistic bacterium against Sclerotium cepivorum causing Allium fistulosum white rot. Total of 146 strains were isolated from A. fistulosum roots. The isolates were screened for antagonism to S. cepivorum and the isolated strain No. AL-1 was selected among these bacteria. It was identified as Serratia plymuthica based on morphological and physiological characteristics according to the Bergey's mannual of systematic bacteriology and 16S rDNA sequences methods. Serratia plymuthica AL-1 showed broad spectrum of antifungal activities against plant pathogenic fungi Alternaria altrata, Colletotrichum gleosporioids, Phoma sp., Rhizoctonia solani, Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium niveum but not inhibited Didymella bryoniae. When S. plymuthica AL-1 cultivated in the TSB medium containing 1% colloidal chitin, the high molecular fraction (>10 kDa) have chitinase activity (3.2 units/ml) and the low molecular fraction (<10 kDa) have not chitinase activity. Oppositely, after heat treatment (80℃ for 30 min) of the cultivation supernatant, the high molecular fractions have not antifungal activity but the low molecular fractions have antifungal activity.

Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-1 Gene (Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정)

  • Yoon, Sang-Hong;Kim, Dong-Gwan;Lee, Young-Han;Shin, Soon-Young;Kwon, Soon-Woo;Lee, Chang-Muk;Kang, Han-Chul;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
    • /
    • v.39 no.2
    • /
    • pp.119-125
    • /
    • 2011
  • The Egr-1 gene is known to be a transcription factor for activating the expression of many tumor-repressing genes. In this study, three strains activating the promoter of the Egr-1 gene were selected, through the use of Egr-1 luciferase reporter assay and western blotting, from amongst approximately 3,800 oligotrophic bacteria isolated from the cultivated soils of various regions within Korea. These strains were identified as Pseudomonas koreensis on the basis of phylogenetic tree analysis of their 16S ribosomal DNA sequences and biochemical characteristics analyses using a variety of commercial kits (API 20NE, ID 32GN, API ZYM kits). In addition, we discovered that these strains produced anti-bacterial activity against Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes.

Molecular phylogeny of Astilbe: Implications for phylogeography and morphological evolution (노루오줌속(Astilbe)의 분자 계통: 계통지리 및 형질 진화에 대한 고찰)

  • Kim, Sang-Yong;Kim, Sung-Hee;Shin, Hyunchur;Kim, Young-Dong
    • Korean Journal of Plant Taxonomy
    • /
    • v.39 no.1
    • /
    • pp.35-41
    • /
    • 2009
  • Astilbe (Saxifragaceae) is a genus well known for its disjunctive distribution in Asia and eastern North America. In this study, we reconstructed a molecular phylogeny of the genus using the sequences of ITS regions of nuclear ribosomal DNA. A total of 17 species representing major lineage of Astilbe and closely related taxa were included in the phylogenetic analyses. We obtained a Bayesian phylogenetic tree in which Saxifragopsis was positioned as a sister group to Astilbe. The Japanese endemic species, A.platyphylla was the most basal lineage within the genus. This species is well known for its distinct morphological features such as unisexual flowers, apetaly, and calyx with 7-11 lobes. Two species, A. biternata, a New World representative of the genus, and A. rivularis widely distributed in S. Asia, branched off early in the evolution of Astilbe. The remaining species formed a strongly supported core clade, which diverged into two robust geographical lineages: the first ("Japonica" clade) of species distributed in Japan, Taiwan, and Philippines and the other ("Rubra" clade), of taxa in China and Korea. The ITS phylogeny indicates that the Bering land bridges were the major route for the origin and dispersal of A. biternata. The two Taiwanese taxa and A. philippinensis were found to derive from the Japanese member, as the genus advanced southwards. The ITS phylogeny suggests that apetaly originated independently at least two times within the genus. Our results do not support Engler's classification system of the genus based on the leaf type (simple vs. compound), but reaffirm Hara's taxonomic idea which primarily considered the features of calyx.

Effect of Disodium Fumarate on In vitro Rumen Fermentation of Different Substrates and Rumen Bacterial Communities as Revealed by Denaturing Gradient Gel Electrophoresis Analysis of 16S Ribosomal DNA

  • Mao, S.Y.;Zhang, G.;Zhu, W.Y.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.4
    • /
    • pp.543-549
    • /
    • 2007
  • Two experiments were conducted to investigate the effects of disodium fumarate on the in vitro rumen fermentation profiles of different substrates and microbial communities. In experiment 1, nine diets (high-forage diet (forage:concentrate, e.g. F:C = 7:3, DM basis), medium-forage diet (F:C = 5:5, DM basis), low-forage diet(F:C = 1:9, DM basis), cracked corn, cracked wheat, soluble starch, tall elata (Festuca elata), perennial ryegrass and rice straw) were fermented in vitro by rumen microorganisms from local goats. The results showed that during 24 h incubations, for all substrates, disodium fumarate increased (p<0.05) the gas production, and tended to increase (p<0.10) the acetate, propionate and total VFA concentration and decrease the ratio of acetate to propionate, whereas no treatment effect was observed for the lactate concentration. The apparent DM loss for tall elata, perennial ryegrass and rice straw increased (p<0.05) with the addition of disodium fumarate. With the exception of tall elata, perennial ryegrass and rice straw, disodium fumarate addition increased the final pH (p<0.05) for all substrates. In experiment 2, three substrates (a high-forage diet, a medium-forage diet and a high concentrate diet) were fermented by mixed rumen microbes in vitro. A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique was applied to compare microbial DNA fingerprints between substrates at the end of 24 h incubation. The results showed that when Festuca elata was used as substrate, the control and disodium fumarate treatments had similar DGGE profiles, with their similarities higher than 96%. As the ratio of concentrate increased, however, the similarities in DGGE profiles decreased between the control and disodium fumarate treatment. Overall, these results suggest that disodium fumarate is effective in increasing the pH and gas production for the diets differing in forage: concentrate ratio, grain cereals and soluble starch, and in increasing dry matter loss for the forages (tall elata, perennial ryegrass and rice straw) in vitro, whereas its effect on changes of ruminal microbial community may largely depend on the general nature of the substrate.