• Journal of Soil and Groundwater Environment
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• v.14 no.3
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• pp.57-67
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• 2009

In soil and sediment environment, microorganisms play major roles in biochemical cycles of ecological significant elements. Because of its ecological significance, microbial diversity and community structure information are useful as indexes for assessing the quality of subsurface ecological environment and bioremediation. To achieve more accurate assessment, it is requested to gain sufficient yield and purity of DNA extracted from various soil and sediment samples. Although there have been a large number of basic researches regarding soil and sediment DNA extraction methods, little guideline information is given in literature when choosing optimal DNA extraction methods for various purposes such as environmental ecology impact assessment and bioremediation capability evaluation. In this study, we performed a thorough literature review to compare the characteristics of the current DNA extraction methods from soil and sediment samples, and discussed about considerations when selecting and applying DNA extraction methods for environmental impact assessment and bioremediation capability evaluation. This review suggested that one approach is not enough to gain the suitable quantity and yield of DNA for assessing microbial diversity, community structure and population dynamics, and that a careful attention has to be paid for selecting an optimal method for individual environmental purpose.

• Applied Biological Chemistry
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• v.39 no.5
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• pp.403-408
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• 1996

Total bacterial community DNA, which was extracted from microcosm soil and field soil after 2,4-D amendments, was analyzed on Southern blots, using the tfdA gene probe derived from plasmid pJP4 and the Spa probe from Sphingomonas paucimobilis. Southern blot analyses with total bacterial DNA extracted from soils Inoculated with Pseudomonas cepacia/pJP4 revealed that DNA probe method could detect the 2,4-D degrading bacteria down to $10^5\;cells/g$ dry soil. In the microcosm experiment, there was a good correlation between 2,4-D degradation and banding patterns in hybridization analyses performed after each 2,4-D treatment using the two probes. When bacterial DNA extracted from microcosm soil was hybridized with the Spa probe, a change in the position of hybrid bands was observed over time in a Southern blot, suggesting that population change or possibly genetic rearrangement in 2,4-D degrading microbial populations occurred in this soil. With the Spa probe, one hybrid DNA band was persistently observed throughout the five 2,4-D additions. When bacterial DNA isolated from the field soil was probed with the tfdA and Spa, strong hybridization signal was observed in the 100 ppm-treated subplot, weak signal In the 10 ppm-treated subplot, and no significant signal in the 1 ppm-treated and control subplots. The data show that DNA probe analyses were capable of detecting and discriminating the indigenous 2,4-D degrading microbial populations in soil amended with 2,4-D under laboratory and field conditions.

• Korean Journal of Environmental Biology
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• v.17 no.3
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• pp.279-284
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• 1999

To investigate soil bacterial diversity according to vegetation types, directly extracted DNA from 5 different soils were cross-hybridized with each other as a probe and target. Pinus densiflora soil was shown the highest value then agricultured soil＞naked soil＞grass soil＞Quercus mongolicas soil in the order of diversity. Cluster analysis by similarity showed that soil microbial communities were categorized into three groups.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.72-77
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• 1999

To investigate soil bacterial diversity according to vegelalioo types, utilizing ability of sole carbon sources and restriction enzyme patterns of 16s rDNA were analyzed. From the both results; five kinds of soil microbial communities were grouped as forest soil (Quercus mongolica and Pinus densi&ra vegetation), grass-agricultured soil and microbial communities of naked soil. But, both soil microbial communities of directily exlracted from ths soil and indirectly extracted from heterotrophic bacteria that cultured soil in LB medium showed very different similarity.

• Korean Journal of Microbiology
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• v.39 no.3
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• pp.187-191
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• 2003

In an attempt to isolate myxobacteria from soil samples, we isolated swarm and fruiting body forming bacteria that have bacteriolytic activity on Coli-spot agar plate. For the classification of myxobacteria, 16S rDNA RFLP patterns were analyzed. Amplified 16S rDNAs of myxobacteria type strains (Family I, II, III and IV), negative control strains and soil-isolates were restricted with HaeIII, EcoRI and EcoRV, respectively. We found that the soil-isolates belongs to myxobacteria Family I, II, III.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.405-408
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• 2010

Diversity of myxobacteria in five soil samples from Asansi and Uponeup in Korea was explored by means of polymerase chain reaction (PCR) using primers that specifically bind 16S rDNA of myxobacteria. DNA sequence analysis of 76 PCR fragments containing myxobacterial 16S rDNA revealed five putative novel myxobacterial genera whose 16S rDNA sequences shared <95% sequence identity with those of the type strains. This finding indicates the presence of many uncultured and unidentified myxobacterial species in Korean soil.

• Proceedings of the Korean Society of Organic Agriculture Conference
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• 2009.12a
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• pp.302-302
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• 2009

유기농업에서 유기물은 양분의 공급, 토양의 이화학성 개선, 토양의 생물학적 건전성 유지 등 중요한 역할을 한다. 토양의 생물학적 건전성은 토양의 생태계적 기능을 지속적으로 유지시키는 토양미생물이 관여하고 있다. 따라서 본 연구는 유기물의 장기연용에 따른 밭토양 미생물의 다양성을 비교 분석하였다. 여러 가지 유기자원을 동일한 기준으로 매년 동일 장소에 처리하였다. 사용된 유기자원은 가축분퇴비, 채종유박인 유기질비료, 볏짚으로만 퇴비화한 볏짚퇴비와 겨울철 휴한기에 헤어리베치를 재배하여 이듬해 봄에 예취한 후 토양에 환원한 녹비처리구, NPK구, 가축분퇴비를 혼용처리한 NPK퇴비군, 양분을 전혀 시용하지 않은 무비구 등 총 7처리구였다. 각각의 처리구에서 토양(0-20 cm)을 채취하여 배양성 토양미생물은 희석평판법으로 해당 선백배지에 시료를 도말 하여 조사하였고 비배양성 미생물은 토양으로부터 genomic DNA를 추출하여 세균의 16S rDNA를 증폭시킨 후 denaturing gradient gel electrophoresis (DGGE)를 수행하여 분석하였다. 주요결과를 요약하면 밭토양에 서식하는 토양미생물의 균수는 처리별간의 차이를 보였으며 유기물처리구가 화학비료처리구보다 높았다. DGGE 분석을 통해 유기물 처리에 따른 군집의 다양성을 살펴본 결과 Fig. 1에서 보는바와 같이 Gel 상에서 다양한 위치의 밴드를 확인할 수 있었고 처리별로 특이 밴드가 있음을 확인할 수 있었다. Fig. 1에서 얻은 DGGE profile상의 밴드 강도와 수를 비교하여 Fig 2와 같은 dendrogram을 나타낼 수 있었다.

• Journal of Applied Biological Chemistry
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• v.56 no.2
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• pp.95-100
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• 2013

Soil microbes are important integral components of soil ecosystem which have significant and diverse role in organic matter decomposition, nitrogen cycling, and nitrogen fixation. In this study an effective denaturing gradient gel electrophoresis (DGGE) method was employed for paddy soil microbial diversity survey. For optimum paddy soil microbial DNA extraction, different methods such as Lysis buffer, skim milk bead, sodium phosphate buffer, Epicentre Soil Master DNA extraction kit (Epicentre, USA) and Mo Bio Power Soil DNA kit (MO BIO, USA) methods were utilized. Among all the method, using Mo Bio Power Soil kit was most effective. DGGE analysis of Bacteria was carried out at 6% polyacylamide gel and 45-60% denaturing gradient in the optimal conditions. Whereas DGGE analysis of fungi was done at 6% polyacrylamide gel and 45-80% denaturing gradient in the optimal conditions. By applying the above assay, it was found that variation within the microbial community of paddy soil occurs by a factor of time. DGGE assay used in this study through for a variety of soil microbial analysis suggests the potential use of this method.

• Korean Journal of Soil Science and Fertilizer
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• v.33 no.4
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• pp.266-274
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• 2000

Understanding of microbial community structure in soil-root system is necessary to use beneficial soil and rhizosphere microbes for improvement of crop production and biocontrol. The knowledge of behavior and function of microbes in soil-root system plays a key role for the application of beneficial inocula. Because the majority of the intact bacteria in soil are unable to grow on nutrient media, both culturable and nonculturable bacteria have to be studied together. In our study, culture-independent survey of bacterial community in the soil-root system of red pepper fields was conducted by the sequence analysis of three universal clone libraries of genes which code for small-subunit rRNA (rDNA). Universal small subunit rRNA primers were used to amplify DNA extracted from each sample and PCR products were cloned into pGEM-T. Out of 27 clones sequenced, 25 clones were from domain bacteria. Two of the rDNA sequences were derived from eukaryotic organelles. Within the domain bacteria, several kingdoms were represented : the Proteobacteria (16 clones). Cytophyga-Flexibacter-Bacteroides group (2 clones). the high G+C content gram-positive group(1 clone) and 4 unknown clones.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1996.11a
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• pp.118-121
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• 1996

환경이 나이어린 학문 정도로 생각하는 것은 환경분야의 눈부신 발전과 연구를 모르는 데서 비롯된 오해이다. 산업화에 따른 산업재앙의 위기에. 처한 환경 선진국들은 이미 막대한 재정지원을 통해서 이미 오래전부터 최신 기법들을 이용한 실험들을 성공리에 마치고 있고 또 실제 생활에 활용하고 있다. 특히 환경오염에 대한 처리방법에 있어서 미국을 비롯한 선진국들은 유전공학을 이용한 최신기법들을 연구개발하고 있으며 이것은 매우 놀랍고 고무적인 일이다. 그런 의미에서, 이 글은 환경오염의 생물학적 처리복구기술에 있어서 DNA probe(DNA 탐침자)의 역할과 이 연구를 통한 미래환경발전의 전망에 대해 살펴보고자 한다.