• Title/Summary/Keyword: environmental DNA

Search Result 1,807, Processing Time 0.041 seconds

Cloning of a Chitinase Gene of Xanthomonas sp. Isolated from Soil and its Expression in E. coli. (토양에서 분리된 Xanthomonas sp.의 Chitinase 유전자 cloning과 E.coli에서의 발현)

  • Kim, Ho-Sang;Seong, Ki-Young;Eun, Moo-Young;Hwang, Cher-Won
    • Applied Biological Chemistry
    • /
    • v.41 no.2
    • /
    • pp.125-129
    • /
    • 1998
  • Xanthomonas sp. isolated from soil exhibited cell wall lytic activity of Candida albicans and secreted chitinase in chitin media. Especially, the chitinase activity was induced by chitin and reached a maximum level at 3 days culture in chitin media. We constructed genomic library of Xanthomonas sp. using cosmid vector in E. coli. Oligonucleotide probe was synthesized from the consensus sequence corresponding to chitinase active site, which was derived from the comparison of amino acid sequences of bacterial chitinase genes. Using this oligonucleotide probe, we screened the genomic library. By restriction enzyme mapping of the positive clones, we identified 4 independent clones which may contain the chitinase gene. One of the clones, named pXCH1 (1.2 kb insert), was further analyzed. Northern blot analysis indicated that is transcripts, 1 kb and 0.8 kb, were induced by chitin. When the cloned gene was induced by IPTG in E.coli cell, chitinase activity which was secreted onto culture media was not observed. However, when the cell was disrupted by using sonicator and then centrifuged, the supernatant exhibited chitinase activity. SDS-PAGE of the supernatant indicated that about 35 kDa protein was induced by IPTG. From these results, it was concluded that the cloned DNA was one of the chitinase genes of Xanthomonas sp.

  • PDF

Improvement of Degrading Activity of Poly(butylene succinateco-butylene adipate)-Degrading Strains Isolated from Soils (토양에서 분리한 Poly(butylene succinate-co-butylene adipate) 분해균의 분해활성 증진)

  • Joo, Hyun-Jin;Kim, Mal-Nam
    • Korean Journal of Environmental Biology
    • /
    • v.27 no.2
    • /
    • pp.198-204
    • /
    • 2009
  • From leaf mold and reclamation site soil of the Capital area of Korea, 3 poly(butylene succinate-co-butylene adipate: PBSA)-degrading strains were isolated through the clear zone test. The PBSA-degrading activities of the strains were assessed by means of a modified Sturm test using 0.01% of PBSA film as a sole carbon source. After the modified Sturm tests for 40 days at the respective isolation temperatures, the 3 strains degraded 30%, 55% and 43% of PBSA, respectively. The isolated strains were identified to be Burkholderia cepacia PBSA-4, Bacillus licheniformisPBSA-5 and Burkholderia sp. PBSA-6 through the 16S rDNA gene sequence analysis. Among them, PBSA-5 degraded both PBSA and Poly(vinyl alcohol). The degradation activity of the PBSA degrading strains appeared to be high at moderate temperatures such as $27^{\circ}C$ and $37^{\circ}C$, and initial inoculum size of $10^{10}cfu\;mL^{-1}$ degraded PBSA 1.2~1.3 more times than that $10^9cfu\;mL^{-1}$. Addition of 0.1 or 0.5% (w/w) of gelatin, yeast extract and ammonium sulfate raised the PBSA degrading activity, and especially addition of 0.1% (w/w) of gelatin enhanced the PBSA degrading activity by more than 33%. The mixed strains degraded PBSA faster than the single strain.

Development of a PCR Assay for Detection of the Protozoan Parasite Perkinsus (PCR 기법을 이용한 바지락포자충 Perkinsus 진단 기술개발)

  • 박경일;박영미;이제희;최광식
    • Korean Journal of Environmental Biology
    • /
    • v.20 no.1
    • /
    • pp.109-109
    • /
    • 2002
  • Detection of protozoan parasites Perkinsus sp. and P. atlanticus was developed in this study using a specific polymerase chain reaction (PCR) to diagnose the presence of those organisms that causes extensive mortalities of marine shellfishes. The PCR was conducted together with fluid thioglycollate medium (FTM) method and 2 M NaOH lysis method. For the test, Manila clams, Ruditapes philippinarum, were collected from four coastal locations in Korea including Wando Island, Gimnyeong, Sungsan and Sogwipo in Jeju. In addition, trophozites of Perkinsus sp. cultivated in vitro and the granular ark clam, Tegillarca granosa, taken from Gangjin on the south coast of Korea, were used as positive and negative controls, respectively. Expected DNA bands were detected in the samples from Wando Island, Sungsan and the in vitro cultured Perkinsus sp. when the probes specific for the genus Perkinsus and P. atlanticus were used. The samples were also positively diagnosed by the FTM and 2 M NaOH methods. In contrast, the Manila clams from Gimnyeong and Sogwipo, and the granular arks clams from Gangjin showed no detectable signs of infection with the PCR, the FTM method and the 2 M NaOH lysis method. On the other hand, being amplified by p. atlanticus specific primer, it is suggested that the protozoan parasite Perkinsus sp. found in the Korean Manila clam is P. atlanticus. Finally the PCR- based assay developed in the present study can be used in detection of Perkinsus infection and discrimination of Peykinsus species in quarantine stations or laboratories due to the high sensitivity and specificity as well as its rapid detection.

Isolation and Characterization of Phosphate Solubilizing Bacteria Pantoea Species as a Plant Growth Promoting Rhizobacteria (식물 생장 촉진 활성을 가진 인산분해 미생물 Pantoea 종의 분리 및 특성 규명)

  • Yun, Chang Yeon;Cheong, Yong Hwa
    • Journal of Life Science
    • /
    • v.26 no.10
    • /
    • pp.1163-1168
    • /
    • 2016
  • Plant growth-promoting rhizobacteria (PGPR) have gained worldwide importance and acceptance due to their agricultural benefits. These microorganisms are potential tools for sustainable agriculture, with effects on plant growth, biofertilization, induced systemic resistance, and biocontrol of plant pathogens. In this study, four different Pantoea species were isolated from field soil, and their plant growth-promoting characteristics were studied. Based on 16S rDNA gene sequencing analyses, the se were grouped into Pantoea ananatis, Pantoea citrea, Pantoea dispersa, Pantoea vagans and named as Pa1, Pc1, Pd1, Pv1, respectively. All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH 5 at 1days after inoculation and production of plant hormone indole acetic acid (IAA) with 85.3±16.3 μg/ml of Pa1, 183.9±16.8 μg/ml of Pc1, 28.8±17.3 μg/ml of Pd1 and 114.1±16.5 μg/ml of Pv1, respectively. Pa1, Pc1 and Pd1 also have high activity for production of gibberellin (GA3) hormone with 331.1±19.2 μg/ml of Pa1, 288.5±16.8 μg/ml of Pc1, 309.2±18.2 μg/ml of Pd1, but Pv1 does not. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants at the range around 32~37% for fresh weight and 10~15% for shoot length enhancement, so that these microbe could be used as a potential bio-fertilizer agents.

Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
    • /
    • v.27 no.12
    • /
    • pp.1415-1420
    • /
    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

The Genetic Approach on Analyzing the Habitat Characteristics of Fairy Pitta Pitta Nympha Inhabiting Jeju Island, the Korean Peninsula and Taiwan (제주도, 한반도 및 대만 내 팔색조의 생태적 서식특성 분석을 위한 유전적 접근)

  • Kim, Eun-Mi;Jeon, Yeon-Seon;Kim, Se-Jae;Kang, Chang-Wan;Won, Hyun-Kyu;Jeong, Gil-Sang
    • Journal of the Korean Society of Environmental Restoration Technology
    • /
    • v.17 no.1
    • /
    • pp.81-90
    • /
    • 2014
  • A Fairy Pitta is a bird known to breed only in mainland China, Taiwan, Japan and Korea and is listed as Vulnerable in the IUCN Red List. We carried out a DNA analysis to contribute to conserve the genetic diversity of Fairy Pitta. 32 samples were collected at Jeju Island, the Korean Peninsula and Taiwan from 2004 to 2013 and DNA was extracted from them and several sequences were amplified-it through PCR. And then we performed the population genetic analysis. We found there was a transversion between nucleotide sequences at CO1 gene, while there was no changes at Cyt-b gene. And we confirmed the polymorphism from two genes was caused from genetic drift not from selection. Through this analysis, the group within the Peninsula was found bigger than other two groups based on the analysis of CO1 gene, and the group from Taiwan was found bigger than other two groups through the analysis of Cyt-b gene. The population genetic structure of mitochondria gene of three group was showing CO1 gene had 5 haplotypes and Cyt-b gene had 6 haplotypes. Haplotype 2 in CO1 gene was found in three group and many individuals of samples had this haplotype. Like CO1 gene, haplotype 2 in Cyt-b gene was found in three group and was included in plenty of individuals. Other haplotypes were not overlaped and broke off among the three groups. To prevent from the extinction of Fairy Pitta and to obtain the genetic diversity, we need to compare with other regional group such as Japan, China and perform additional research in the non-breeding area.

Molecular and Ecological Analyses of Microbial Community Structures in Biofilms of a Full-Scale Aerated Up-Flow Biobead Process

  • Ju, Dong-Hun;Choi, Min-Kyung;Ahn, Jae-Hyung;Kim, Mi-Hwa;Cho, Jae-Chang;Kim, Tae-Sung;Kim, Tae-San;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.2
    • /
    • pp.253-261
    • /
    • 2007
  • Molecular and cultivation techniques were used to characterize the bacterial communities of biobead reactor biofilms in a sewage treatment plant to which an Aerated Up-Flow Biobead process was applied. With this biobead process, the monthly average values of various chemical parameters in the effluent were generally kept under the regulation limits of the effluent quality of the sewage treatment plant during the operation period. Most probable number (MPN) analysis revealed that the population of denitrifying bacteria was abundant in the biobead #1 reactor, denitrifying and nitrifying bacteria coexisted in the biobead #2 reactor, and nitrifying bacteria prevailed over denitrifying bacteria in the biobead #3 reactor. The results of the MPN test suggested that the biobead #2 reactor was a transition zone leading to acclimated nitrifying biofilms in the biobead #3 reactor. Phylogenetic analysis of 16S rDNA sequences cloned from biofilms showed that the biobead #1 reactor, which received a high organic loading rate, had much diverse microorganisms, whereas the biobead #2 and #3 reactors were dominated by the members of Proteobacteria. DGGE analysis with the ammonia monooxygenase (amoA) gene supported the observation from the MPN test that the biofilms of September were fully developed and specialized for nitrification in the biobead reactor #3. All of the DNA sequences of the amoA DGGE bands were very similar to the sequence of the amoA gene of Nitrosomonas species, the presence of which is typical in the biological aerated filters. The results of this study showed that organic and inorganic nutrients were efficiently removed by both denitrifying microbial populations in the anaerobic tank and heterotrophic and nitrifying bacterial biofilms well-formed in the three functional biobead reactors in the Aerated Up-Flow Biobead process.

Occurrence of Gray Mold Caused by Botrytis cinerea on Okra in Korea (Botrytis cinerea에 의한 오크라 잿빛곰팡이병)

  • Choi, JangNam;Choi, InYoung;Lee, KuiJae;Lee, JungNo;Cho, SeongWan;Shin, HyeonDong;Galea, Victor
    • Research in Plant Disease
    • /
    • v.24 no.4
    • /
    • pp.302-307
    • /
    • 2018
  • From 2014 to 2016, approximately 5% of okra fruit were observed displaying gray mold symptoms at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. The symptoms observed were water-soaked, brown or gray spots, and abundant mycelial with conidia appearing on the infected fruit. Initial infection commenced from the base of fruit and gradually moved to the pod, where it finally resulted in collapse. Colonies on potato dextrose agar were gray to grayish brown, felted and cottony expanding 65-80 mm after one week. The fungus formed several black sclerotia ranging $1.0-3.5{\times}0.5-3.0mm$ on the Petri dish after two weeks. The conidia were one-celled, ellipsoidal or ovoid, colorless or pale brown, and $6.2-15.4{\times}5.0-10.4{\mu}m$. Conidiophores arose solitary or in groups, straight or flexuous, septate, with an inflated basal cell brown to light brown, and measured $85-450{\times}10.0-40.0{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of internal transcribed spacer rDNA, the fungus was identified as Botrytis cinerea Pers. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of B. cinerea on okra in Korea.

First Report of an Unrecorded Nematode-trapping Fungus, Arthrobotrys sinensis in Korea (국내 미기록 선충포식성 곰팡이 Arthrobotrys sinensis의 형태 및 분류)

  • Ha, Jihye;Kang, Heonil;Kang, Hangwon;Kim, Donggeun;Lee, Dongwoon;Kim, Yongchul;Choi, Insoo
    • Korean journal of applied entomology
    • /
    • v.58 no.1
    • /
    • pp.9-13
    • /
    • 2019
  • Nematode-trapping fungi use various specialized traps to capture nematodes. A fungus that can capture nematodes in three dimensional adhesive networks was isolated from the soil around the root of Cucumis melo L. (Oriental melon) in Seongju, Korea. The conidiophores were found to be septate, hyaline, erect and $290-528(342.8){\mu}m$ high. It produces obovoid shape and 1-3 septate (commonly 2-septate) conidia with a size of $30.5{\times}20.3{\mu}m$. Molecular analysis of 5.8 S rDNA displayed 99% similarity to Arthrobotrys sinensis. On the basis of morphological, morphometric and molecular studies, the fungus was identified as A. sinensis. It is the first report in Korea which can be one of biological control resource of plant-parasitic nematode.

Distribution and Species Diversity of Wild Yeasts Isolated from Flowers in Korea (국내에 서식하는 꽃에서 분리한 야생 효모 분포 및 종 다양성)

  • Kim, Jeong-Seon;Lee, Miran;Kim, Jae Yoon;Heo, jun;Kwon, Soon-Wo;Yun, Bong Sik;Kim, Soo-Jin
    • The Korean Journal of Mycology
    • /
    • v.48 no.4
    • /
    • pp.475-484
    • /
    • 2020
  • Various indigenous yeasts were isolated and obtained from flowers in the Republic of Korea, and their distribution and species diversity were studied. Seventy-seven flowers were collected from 25 areas in Korea, and 502 yeast strains were isolated from these flowers. A total of 50 species were identified by comparing large subunit rDNA gene sequence homology with the type strains of yeasts. The analysis of yeast distribution showed that the dominant yeast species were Aureobasidium pullulans, A. leucospermi, and Filobasidium magnum in each region and flower samples. Except for the above three yeast species, no species of yeasts showed any meaningful distribution among the habitat regions and sources. In conclusion, 50 species of indigenous yeasts were obtained from flowers that can be used as industrial resources, and the data could be used for further research on yeast diversity and interactions between yeast and its host.