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경작지토양에서 미생물제제가 미생물의 다양성과 고추의 생육에 미치는 영향
안창환 ( Chang Hwan An ),임종희 ( Jong Hui Lim ),김요환 ( Yo Hwan Kim ),정병권 ( Byung Kwon Jung ),김진원 ( Jin Won Kim ),김상달 ( Sang Dal Kim ) 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 2012 한국미생물·생명공학회지 Vol.40 No.1
We investigated the effects on soil microbial diversity and the growth promotion of red pepper resulting from inoculation with a microbial agent composed of Bacillus subtilis AH18, B. licheniformis K11 and Pseudomonas fluorescens 2112 in a red pepper farming field. Photosynthetic bacteria, Trichoderma spp., Azotobacter spp., Actinomycetes, nitrate oxidizing bacteria, nitrite oxidizing bacteria, nitrogen fixing bacteria, denitrifying bacteria, phosphate solubilizing bacteria, cellulase producing bacteria, and urease producing bacteria are all indicator microbes of healthy soil microbial diversity. The microbial diversity of the consortium microbial agent treated soil was seen to be 1.1 to 14 times greater than soils where other commercial agent treatments were used, the latter being the commercial agent AC-1, and chemical fertilizer. The yield of red pepper in the field with the treated consortium microbial agent was increased by more than 15% when compared to the other treatments. Overall, the microbial diversity of the red pepper farming field soil was improved by the consortium microbial agent, and the promotion of growth and subsequent yield of red pepper was higher than soils where the other treatments were utilized.
토양효소활성을 이용한 미생물제제 처리 고추경작지의 토양미생물군집 분석
김요환 ( Yo Hwan Kim ),임종희 ( Jong Hui Lim ),안창환 ( Chang Hwan An ),정병권 ( Byung Kwon Jung ),김상달 ( Sang Dal Kim ) 한국응용생명화학회(구 한국농화학회) 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.1
Increasing concerns over green farming technology, plant growth promoting rhizobacterium (PGRP) having growth promoting as well as plant disease suppressing properties was recently preferred to use for biological control of plant pathogens infecting plant. We measured the influence of the selected microbial consortium agents-a mixture of PGPR strains-, commercial biofungicide, and chemical pesticides on soil microbial community in red pepper field. The activities of soil enzyme such as dehydrogenase, urease, phosphatase, β-glucosidase, and cellulase were analyzed to investigate that of soil microbial community. We also measured plant length, main stem, stem diameter, number of branches and yields of red-pepper in order to observe the red pepper growth promotion. The results of measuring enzyme activities were dehydrogenase 3.5584 μg TPF g-1h-1, urease 15.8689 μg NH4 -N g-1h-1, phosphatase 0.5692 μg PNP g-1h-1, β-glucosidase 2.4785 μg PNP g-1h-1, and cellulase 86.1597 μg glucose g-1h-1 in the soil treated with the microbial consortium agents, so it came out to be very active in the soil. Observing the growth of red-peppers, the main-stem length and the stem diameter were 6.1% and 8.1% higher in the soil treated with the selected microbial consortium agent than the chemical pesticides. After harvesting, yields were 7.3% higher in the soil treated with selected microbial consortium agents than the chemical pesticides. These results showed that microbial consortium agents contribute to increasing soil microbial diversity, growth promoting, and yield of red pepper.
미생물제제시용 고추경작지로부터 식물생장홀몬과 항진균물질을 동시에 생산하는 식물생장촉진근권세균의 선발 및 동정
정병권 ( Byung Kwon Jung ),임종희 ( Jong Hui Lim ),안창환 ( Chang Hwan An ),김요환 ( Yo Hwan Kim ),김상달 ( Sang Dal Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2012 한국미생물·생명공학회지 Vol.40 No.3
In this study, a total of more than 1,000 bacteria, including 739 species of aerobic bacteria, 80 species of urease producing bacteria and 303 species of photosynthetic bacteria, were isolated from red-pepper field soils located in the Gyeongsan Province of the Republic of Korea. Amongst these, 158 species of aerobic bacteria, 70 species of urease producing bacteria and 228 species of photosynthetic bacteria were found to be auxin producing soil bacteria through quantification analysis with the Salkowski test. The latter groupings were then tested for antifungal activities to β-Glucanase and siderophore using CMC congo red agar and CAS blue agar media. In addition, the selected strains were examined for antifungal activity against various phytopathogenic fungi on PDN agar media. Six strains; BCB14, BCB17, C10, HA46, HA143, and HJ5, were noted for their ability to both produce auxin and act as antifungal substances. 16S rDNA sequence comparison analyses of these six strains identified them as Bacillus subtilis BCB14, B. methylotrophicus BCB17, B. methylotrophicus C10, B. sonorensis HA46, B. subtilis HA143, and B. safensis HJ5.
Denaturing gradient gel electrophoresis를 이용한 한국의 논 토양 미생물 다양성 분석 방법
최명은 ( Myeongeun Choe ),홍성준 ( Sung Jun Hong ),임종희 ( Jong Hui Lim ),곽윤영 ( Yun Young Kwak ),백창기 ( Chang Gi Back ),정희영 ( Hee Young Jung ),이인중 ( In Jung Lee ),신재호 ( Jae Ho Shin ) 한국응용생명화학회(구 한국농화학회) 2013 Journal of Applied Biological Chemistry (J. Appl. Vol.56 No.2
현재 토양 생태에서 토양미생물은 유기물 분해, 질소 순환, 식물의 질소 이용 등 중요한 역할을 하고 있어, 토양 내 미생물 다양성을 분석하기 위한 연구는 지속적으로 진행되어 오고 있다. 본 연구에서는 논 토양의 미생물 생태 다양성을 조사하기 위한 효과적인 방법으로 denaturing gradient gel electrophoresis (DGGE)를 적용하고자 본 연구를 수행하였다. 논 토양 미생물의 DNA를 분리하기 위하여 lysis buffer method, skim milk bead method, sodium phosphate buffer method, Epicentre SoilMaster DNA extraction kit (Epicentre, USA), Mo Bio PowerSoil kit (Mo Bio, USA)를 이용하여 토양 내 gDNA 최적 추출방법을 확인하였다. 그 결과 Mo Bio PowerSoil kit를 사용하였을 때 Shannon 다양성지수가 세균 3.3870, 진균 3.6254으로 미생물 다양성 분석시에 가장 효과적이었다. DGGE 분석을 위한 조건은 세균의 경우 6% polyacylamide gel, 45? 60% denaturing gradient였고, 진균의 경우 6% polyacrylamide gel, 45?80% denaturing gradient에서 최적 분석조건을 보였다. 위의 분석법을 적용하여 논 토양내의 미생물 군집의 변화를 살펴보면 시간의 변화 요인에 의해 미생물 변화가 일어나는 것을 알 수 있었다. 본 연구에서 사용된 DGGE 분석법을 통해 논토양 미생물의 분석 가능성을 제시 할 수 있었다. 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 say used in this study through for a variety of soil microbial analysis suggests the potential use of this method.
마늘 잎집썩음병과 마른썩음병을 길항하는 토착길항미생물의 선발 및 기작
김상달 ( Sang Dal Kim ),정희영 ( Hee Young Jeong ),김병극 ( Byung Keuk Kim ),임종희 ( Jong Hui Lim ),이중종 ( Jung Jong Lee ) 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 2010 한국미생물·생명공학회지 Vol.38 No.3
마늘병원균에 대한 토착길항미생물 선발을 위하여 경북 영천시 신덕리 및 구계리 일대 16개소의 토양 시료로부터 160균주의 세균들을 분리하였다. 이들 분리된 세균 중 마늘잎집썩음병의 원인균인 P. marginalis와 마른썩음병의 원인균인 F. oxysporum에 대하여 길항능이 매우 뛰어난 3종의 토착길항세균을 선발하였다. 선발 균주들을 16s rDNA sequencing과 Bergey`s manual을 이용한 방법에 의해 동정한 결과 B. subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240 균주들로 동정할 수 있었다. 선발된 3종의 토착길항세균들은 항세균 및 항진균 길항기작인 siderophore, β-glucanse, chitinase, 항생물질 생산능을 가진 균주들이었으며, 식물의 생장촉진에 도움이 되는 인산가용 화능과 식물생장촉진호르몬인 옥신 및 지베렐린을 생산하는 균주들로 확인되었다. 또한, 3종의 토착길항세균들은 in vivo포트실험을 통해 잎집썩음병원균인 P. marginalis 에 대한 방제율이 70% 이상이고 동시에 생장촉진능도 있다는 것을 검증하였다. 그리고 마른썩음병원균인 F. oxysporum에 대한 방제율도 B. subtilis YC82와 B. amyloliquefaciens Y240의 경우 60%, B. vallismortis YC84는 80% 이상으로 나타났다. 또한, 발아촉진능에서는 선발된 토착길항세균의 처리구가 무처리구에 비하여 20% 이상 향상된 우수한 발아촉진능을 나타내었으며, 생장촉진능 역시 초장 길이의 차이에서 B.subtilis YC82는 평균 49±1.53 mm, B. vallismortis YC84는 평균 47±1.15 mm, B. amyloliquefaciens YC240에서는 평균 80±1.21 mm로 무처리구의 평균 39±1.51mm 보다 매우 우수하였다. 이와 같은 결과는 선발된 마늘잎집썩음병 및마른썩음병 방제용 토착길항세균은 마늘병원균 방제능과 동시에 생장촉진능을 가지는 다기능 토착길항세균으로 향후추가적인 현장적용시험을 통하여 병방제능 및 생장촉진능 검증이 이루어 진다면 마늘병해방제 및 생장촉진용 미생물 제제 개발이 가능할 것이라 생각된다. Sheath rot and dry rot disease caused by Pseudomonas marginalis and Fusarium oxysporum were serious problems in garlic farmland. In this study, total of 160 indigenous antagonistic bacteria were isolated from 16 farmlands in Yeongcheon, Korea. Among these, 15 strains were able to inhibited P. marginalis and F. oxysporum. The 16s rDNA genes of the selected 15 strains were amplified and sequenced. The strains has strong antagonistic ability against garlic pathogens was achieved Bacillus subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240. The selected 3 strains tested for investigation of antifungal mechanisms further analyses; 3 strains of these validated for production of siderophore, β-glucanase and chitinase using CAS (chrome azurol S) blue agar, CMC-congo red agar and DNS method. The 3 strains were able to utilized insoluble phosphate as dertermined by vanado-molybdate method. The 3 strains verified for production of auxin and gibberellic acid using Salkowski test and holdbrook test. Also, 3 strains showed stimulation germination, stem growth promoting activity on the in vivo test. The 3 strains were able to effectively suppress P. marginalis and F. oxysporum causing sheath rot and dry rot diseases on the in vivo pot test.