초록 ▼

◦ 질소고정균 50균주 분리·동정
◦ 우수 질소고정균 13종 선발하여, 미생물은행에 11종 기탁
◦ 최적대량 배양 조건을 확립하여, 고활성 질소고정균 4종을 선발하였고 미생물 제제를 제조하였음
◦ 각각 다른 60개의 sampling site에서 질산화균 enrichment를 수행하여, 질산화균 증식을 PCR 등 방법으로 확인함
◦ 최적 배양법으로 질산화가 뛰어난 culture를 선택하고,RT-PCR을 활용하여 cell counting하여 유효성을 검증함
◦ kit를 이용하여 질산화를 확인하였고, 활성이 뛰

◦ 질소고정균 50균주 분리·동정
◦ 우수 질소고정균 13종 선발하여, 미생물은행에 11종 기탁
◦ 최적대량 배양 조건을 확립하여, 고활성 질소고정균 4종을 선발하였고 미생물 제제를 제조하였음
◦ 각각 다른 60개의 sampling site에서 질산화균 enrichment를 수행하여, 질산화균 증식을 PCR 등 방법으로 확인함
◦ 최적 배양법으로 질산화가 뛰어난 culture를 선택하고,RT-PCR을 활용하여 cell counting하여 유효성을 검증함
◦ kit를 이용하여 질산화를 확인하였고, 활성이 뛰어난 액상 농축 배양액으로 미생물 제제를 제조함
◦ 탈질균 30균주이상 분리·동정
◦ 분리한 균주 130여 종에서 탈질활성을 갖는 균주를 선별하기 위해, PCR, durham test, kit, RT-PCR 방법을 이용함
◦ 우수한 균주를 20종을 선발하여, 미생물은행에 기탁
◦ 저농도부터 고농도의 NO3 배지에서 탈질 테스트를 진행하여 최종 5균주를 선발하였고, 최적의 대량배양을 통해 균주를 회수하고 미생물 제제를 제조함

(출처: 요약서 4p)

Abstract ▼

□ Purpose&Contents
◦ Isolation and identification of 50 isolates of nitrogen-fixing bacteria
- Several different genus of nitrogen-fixing bacteria (Rhizobium, Mesorhizobium, Bradyrhizobium, Azotobacter, Beijerinckia, Bacillus etc.) and several types bacteria to be secured were grown in a nitro

□ Purpose&Contents
◦ Isolation and identification of 50 isolates of nitrogen-fixing bacteria
- Several different genus of nitrogen-fixing bacteria (Rhizobium, Mesorhizobium, Bradyrhizobium, Azotobacter, Beijerinckia, Bacillus etc.) and several types bacteria to be secured were grown in a nitrogen-free medium and identified as a 16S rDNA base sequence
◦ Isolation and identification of 30 isolates of denitrifying bacteria
◦ Selection of high activity strains through active measurement of nitrogen fixation ability, nitrifying ability and denitrifying ability
- Classification of novel strains of bacteria through polyphatic analysis(16S rDNA sequence, phylogenetic tree, G+C contents, whole cell fatty acid, quinone type, polar lipid, various substrate tests, SEM or TEM for shape observation)
◦ Validation of nitrogen-fixing bacteria and nitrous oxide bacteria through crop testing
◦ Exploring the Optimal Conditions of Mass Cultivation of Superior Nitrogen Cycle-related Bacteria
- Searching the optimal conditions for strains to grow best while controlling the nutrition concentration, pH condition, nutrient salts, and airvolume of the growth medium using a 10 L fermenter
◦ Development of Microbial Agent by mixing activated Nitrogen fixing bacteria
◦ Development of Microbial Agent through liquid mixture of nitrifying bacteria
◦ Development of Microbial Agent by mixing activated denitrifying bacteria

□ Results
◦ Isolation of 50 strains of nitrogen fixation
◦ Select 13 isolates of superior nitrogen fixation bacteria and deposit 11isolates with culture collection
◦ 4 strains of highly active nitrogen-fixing bacteria were selected and microbial agents were manufactured by establishing conditions for optimal mass incubation.
◦ While nitrifying bacteria are incubated, the growth of nitrifying bacteria were checked by PCR etc. in different 60 site of enrichment cultures.
◦ Select the enrichment culture with excellent activity as the optimum incubation method. Cell counting is performed using RT-PCR to verify validity.
◦ The nitrifying activity was checked using the kit, and microbial agents were prepared with a liquid enriched culture with high activity.
◦ Isolation of 30 or more denitrifying bacteria
◦ PCR, Durham test, Kit, and RT-PCR method are used to select strains that are denitrifying activated in over 130 isolates of separated strains.
◦ Select 20 activity strains and deposit them with culture collection
◦ The final five strains were selected through denitrification tests on low to high concentrations of NO3 growth media, and the strains wererecovered through optimum mass culture and microbial agents were manufactured.

□ Expected Contribution
◦ The efficacy evaluation of nitrogen fixation bacteria has been completed, and the effects of more various plants are further studied and used as microbial agents.
◦ The activity was confirmed by establishing the optimal incubation condition for the efficient incubation of nitrifying bacteria, and based on this, it will be evaluated by creating microbial agents for liquid nitrifying bacteria, such as sewage treatment plants.
◦ Using RT-PCR technology, which was introduced to verify the activity of enriched nitrifying bacteria culture, the company is planning to establish a service to detect target bacteria present in certain specimens,such as environmental samples and sludge, through cell counting.
◦ If there is a process that lacks nitrogenous bacteria during the sewage treatment plant process, a system is also planned to be prepared to supply bacteria through cell counting and optimum incubation conditions.

(출처 : SUMMARY 7p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 4
• 국 문 요 약 문 ... 5
• Summary ... 7
• 목차 ... 9
• 제 1 장 연구 개발 과제의 개요 ... 10
• 제1절 연구 개발 목적 ... 10
• 제2절 연구 개발의 필요성 ... 10
• 제3절 연구 개발 범위 ... 11
• 제 2 장 연구 수행 내용 및 결과 ... 12
• 제 3 장 목표달성도 및 관련분야 기여도 ... 46
• 제1절 : 목표대비 달성도 ... 46
• 제2절 : 정량적 성과(논문게재, 특허출원, 기타)를 기술 ... 47
• 제 4 장 연구 결과의 활용 계획 ... 48
• 제 5 장 연구 개발 결과의 보안 등급 ... 49
• 제 6 장 국가과학기술지식정보서비스에 등록한 연구시설·장비 현황 ... 50
• 제 7 장 연구개발과제의 대표적 연구실적 ... 51
• 제 8 장 기타사항 ... 53
• 제 9 장 참고문헌 ... 54
• 끝페이지 ... 57