초록 ▼

□ 과제명 : 농업용 미생물의 현장적용기술 개발
▶ 연구목적 :
○ 지자체 및 민간에서 활용하고 있는 미생물제의 효능검증 및 활용증진
○ 미생물비료 및 생물활성제용 토착미생물 선발하여 미생물유래 기능성
물질 탐색 및 활용방법 개발 및 농업용 미생물자원 인벤토리 구축
▶ 주요연구내용 :
○ 제1세부과제 : 유용미생물의 효능평가 및 현장활용 증진기술 개발
농업미생물의 현장활용성 증대를 위하여 현장 핵심인력을 대상으로 워크숍을 실시하여 현장 기술수요를 파악하고, 지자체에서 생산보급하고 있는 미생물(B

□ 과제명 : 농업용 미생물의 현장적용기술 개발
▶ 연구목적 :
○ 지자체 및 민간에서 활용하고 있는 미생물제의 효능검증 및 활용증진
○ 미생물비료 및 생물활성제용 토착미생물 선발하여 미생물유래 기능성
물질 탐색 및 활용방법 개발 및 농업용 미생물자원 인벤토리 구축
▶ 주요연구내용 :
○ 제1세부과제 : 유용미생물의 효능평가 및 현장활용 증진기술 개발
농업미생물의 현장활용성 증대를 위하여 현장 핵심인력을 대상으로 워크숍을 실시하여 현장 기술수요를 파악하고, 지자체에서 생산보급하고 있는 미생물(Bacillus subtilis, 유산균, 광합성균, 효모 등)을 수집하여 항균활성, 질소고정능력, 인산가용화 능력, 오옥신 생성여부 및 효소활성을 분석하고 그 결과를 정리하여 지자체 미생물 담당자 대상 워크숍에서 발표하였고, 지자체 생산 보급 미생물의 저장조건(온도, 기간)에 따른 밀도 변화를 구명하였다. 또한 오이, 고추 등 작물에 대한 생장촉진효과를 구명하는 선발된 균주를 공시하여 한편 고추 및 토마토 흰가루병에 대한 방제효과를 조사하였다. 포장 조건에서 우수한 방제효과를 보인 Bacillus subtilis R2-1을 흰가루병 방제제로 특허출원하였다(10-2011-0077891)
○ 제2세부과제 : 다기능 미생물의 농가실용화 기술 개발
유기물 배지로부터 다양한 미생물을 분리하여 다양한 기능을 갖는 미생물을 선발하였고 그 중 2종의 미기록종(Pandoraea thiooxydans sp. nov. 와 Burkholderia kukuriensis subsp. thiooxydans) NCBI에 등재하는 한편 국외논문에 게재하였다. 다기능 미생물의 대량배양을 위하여 저렴하고 미생물생육에 우수한 유기물원을 선발하여 3종의 미생물 배양배지를 개발하였다. 그람양성균 배양을 위해서는 생선아미노산액비, 휴믹산 및 당밀을 첨가한 배지와 혼합곡, 천일명 및 당밀을 첨가한 배지를 선발하였고, 그람음성균 배양을 위해서는 유채박, 휴믹산과 당밀을 첨가한 배지를 개발하였다. 또한 상추, 배추, 고추 생산성을 촉진하는 다기능 미생물로 Bacillus subtilis R2-1과 Pantea ananatis를 선발하였다. 또한 Bacillus subtilis R2-1을 처리할 경우 토마토 흰가루병 방제효과가 우수하였다.
○ 제3세부과제 : 미생물비료자원 개발을 위한 농경지미생물의 다양성 평가
유기농경지 군집의 다양성 평가를 위한 균특이적 프라이머를 활용한 DGGE 분석법을 개발하였다. 이 분석법을 활용하면 유기물 처리간 유전적 차이(DNA 밴드패턴)를 확인할 수 있었다. 유기물 장기연용포장의 토양미생물 다양성을 분석한 결과 녹비와 같은 유기물을 처리한 토양에서 미생물 다양성이 향상되었으며 유기물 처리별 미생물 군집의 종 다양성 및 풍부도가 증가되었다. 유기물 처리에 따른 세균 군집 cluster를 그룹화가 가능하였다. 세균 군집 cluster는 녹비, 가축분, 볏짚퇴비처리구가 제1그룹으로, N-P-K+가축분, 무비, NPK 처리구가 제2그룹으로 그리고 유박처리구가 제3그룹으로 구분되었다.
○ 제4세부과제 : 선발 미생물 자원의 기능평가 및 인벤토리 구축
농경지 토양미생물 기능평가에 의한 농업환경보전 및 미생물비료 개발을 위해 논토양, 밭토양, 과수원토양 등으로부터 세균 500여 균주를 분리하고 12가지 탄소원에 대한 대사능력을 검정하고 지방, 단백질, 키틴, 섬유질 분해효소에 대한 활성과 인산가용화 능력을 평가하였다. 분리된 균의 동정을 위해 지방산분석과 16S rRNA 유전자를 이용하였으며, 이들을 종합하여 인벤토리를 구축하였다. 더불어 비료연용지 토양에 대한 분석결과, 3요소(질소, 인산, 칼리)로 처리된 토양이 무처리된 토양보다 미생물활성이 더 높게 나타났다. 또한 3요소로 처리된 토양이 규소, 석회 또는 퇴비로 처리된 토양보다 낮은 대사능력을 보이는 것으로 볼 때 토양개량제의 추가적인 사용이 미생물의 활성을 더 높게 하는 것으로 판단된다. 시설재배지 토양을 분석한 결과, 유기농 토양 미생물군집이 관행농 토양보다 더 높은 탄소대사능력을 보였으며 보다 더 다양한 속의 세균을 분리할 수 있었다. 추후 선발한 자원에 대한 인벤토리는 유용미생물의 검정 등에 활용할 계획이며, 배양법이 아닌 비배양법으로 유기농과 관행농 토양 미생물에 대한 다양성을 평가할 계획이다.
○ 제5세부과제 : 농업미생물 이용 유기농가의 경영성과 분석
2009년 말 현재 우리나라에서 생물농약으로 등록된 자재는 살균제, 살충제, 제초제를 합해 모두 31종류이다. 이 가운데 수입된 자재가 12종류, 국내에서 제조된 자재가 19종류로 최근 수입 생물농약이 증가하고 있는 추세에 있다. 종류별로 보면 살균제가 17개, 살충제 13, 제초제 1종류로 살균제가 가장 많은 수를 차지하고 있다. 한편 2008년도에 국내에서 등록된 생물농약의 출하량은 49MT, 출하금액은 1,780백만원으로 전체 농약출하금액의 0.15%를 차지하고 있다. 2006년도 생물농약의 국내 시장규모는 약 370억원 정도로 추산되며, 전체 친환경농업의 재배면적과 생물농약의 살포 회수 등을 고려해 볼 때 2010년도 생물농약 시장규모는 약 820억원 정도가 될 것으로 예측되고 있다.

Abstract ▼

In order to improve practical utility of agro-microorganisms(AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antif

In order to improve practical utility of agro-microorganisms(AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and pectinase. In addition, their plant promotion effect was evaluated on tomato and pepper. As a result of analysis of 16S-rRNA sequences and fatty acid by MIDI system on 47 AMs, we found that 8 isolates of them were mis-identified. It was confirmed that Bacillus species and Paenibacilus species had antifungal activity against several plant pathogens and activity of siderophore formation, IAAs formation and nitrogen fixation.
Lactic acid bacteria most had mostly strong activity of phosphorus solubilization and pectinase and partly activity of antagonism, and siderophore and IAAs formation. Yeasts had partly antifungal activity, IAAs formation and pectinase activity. Photosynthetic bacteria showed a little activity of siderophore formation and nitrogen fixation. Growth promoting effect of 47 AMs were evaluated on cucumber and tomato. In cucumber, 23AMs promoted root growth, but did not promote shoot growth. In tomato, 24AMs promoted shoot growth and 5AMs did root growth. As a result of investigation of root colonization of 47AMs on tomato using root imprint method, 22 AMs including GH-LAB-1 showed high colonization activity. Control effect of AMs on red pepper powdery mildew and cucumber powdery mildew were investigated.
KP-Y-1 and CW-PBS showed high control effect against red pepper powdery mildew and BS R2-1 did comparative high control effect against tomato powdery mildew. BS R2-1 isolate showed high control effect on organically cultivated red pepper in open field. In vitro test suppressive effect of AMs on red pepper gray mold using wounded fruits and tomato gray mold using cut stems, 11 AMs including GH-PSB-1 showed control effect against tomato gray mold and 22 AMs including GH-LAB-1 did control effect against red pepper gray mold. To develop the functional media for promoting plant growth, several organic matter media including Panchakavyia were evaluated for plant promotion effect on radish and Chinese cabbage and multi-function like siderophore formation, nitrogen fixaton, phosphorous solubilization, IAA formation, enzyme activity, etc. 33 times-diluted Panchakavyia promoted root growth of Chinese cabbage and shoot growth of radish, showed siderophore formation, nitrogen fixation, phosphorous solubilization, ACC deaminase activity, IAA formation, enzyme activity(pectinase, cellulase). Among several organic matter media(OMM), OMM supplemented with Bacillus subtilis R2-1 promoted the highest growth of Chinese cabbage and radish, showed siderophore formation, nitrogen fixation, phosphorous solubilization, ACC deaminase activity, IAA formation, enzyme activity(pectinase, cellulase). Fourteen bacteria with multi-functions were finally selected from OMM and recorded on NCBI(National Center for Biotechnology Information) list.
Two mass-culturing media of agro-microorganisms were developed for culture Gram positive bacterium, Bacillus subtilis. The composition of the two media were as follows: Aminoacid liquid manure medium(amino acid manure 0.1%, humic acid 1.0% and molasses 1.0%) and mixed grain-natural salt medium(mixed grain 0.5%, natural salt 1.0% and molasses 1.0%). One mass-culturing media of agro-microorganisms were developed for culture Gram negative bacterium, Pantoea ananatis. The composition of the media was as follows: Vegetable seed cake medium(vegetable seed cake 1.0%, humic acid 1.0%, molasses 1.0%).
As a result of seed germination test by treatment of the two selected bacteria, Bacillus subtilis R2-1 and Pantoea ananatis B1-9, with multi-functions, while seed germination of red pepper were suppressed a little by Bacillus subtilis R2-1, the other seeds were germinated well.
Treatment of Bacillus subtilis R2-1 did not have affect on growth of lettuce, Chinese cabbage and red pepper. However treatment(106cfu/㎖) of Pantoea ananatis B1-9 promoted the growth of lettuce(leaf length and fresh weight) and Chinese cabbage(leaf length) and did not do the growth of red pepper in the green house. Bacillus subtilis R2-1 did not promote growth of lettuce, but Pantoea ananatis B1-9 promoted remarkably that of lettuce. There is a difference in plant promotion effect Bacillus subtilis R2-1 and Pantoea ananatis B1-9 according to kind of red pepper cultivars. While Bacillus subtilis R2-1 and Pantoea ananatis B1-9 promoted plant growth of red pepper in PR Daechon, only Pantoea ananatis B1-9 did plant growth of red pepper in Shindokbuljanggun.
The two microorganisms, Bacillus subtilis R2-1 and Pantoea ananatis B1-9, suppressed powdery mildew of tomato. Control effect of Bacillus subtilis R2-1 depended on cell concentration. We thought that mode of action of Bacillus subtilis R2-1 for controlling powdery mildew of tomato be antibiosis. Meanwhile control effect of Pantoea ananatis B1-9 was not correlated with cell concentration. We thought that mode of action of Pantoea ananatis B1-9 for controlling powdery mildew of tomato to be antibiosis and need further study.
Organic and chemical fertilizer amendments are an important agricultural practice for increasing crop yields. In order to maintain the soil sustainability, it is important to monitor the effects of fertilizer applications on the shift of soil microorganism, which control the cycling of many nutrients in the soils. Here, culture-dependent and culture- independent approaches were used to analyze the soil microorganism and community structure under six fertilization treatments, including green manure, rice straw compost, rapeseed cake, pig mature compost, NPK +pig mature compost, NPK and control. Both organic and chemical fertilizers caused a shift of the cultural microorganism CFUs after treatments. Bacterial CFUs of the organic fertilization treatments were CFUs after treatments. Bacterial CFUs of the organic fertilization treatments were significantly higher than that of chemical fertilization treatments. The DGGE profiles of the bacterial communities of the samples showed that the green manure treatment was a distinct difference in bacterial community, with a greater complexity of the band pattern than other treatments. Cluster analyses based on the DGGE profile showed that rice straw compost and pig mature compost had a similar banding pattern and clustered together firstly. Rapeseed cake, NPK, NPK + pig manure compost and control clustered together in other sub-cluster and clearly distinguished from green manure. Bacterial community composition were analyzed using 16S rRNA gene-based pyrosequencing for long term application of organic material in upland soil. The relative ration of the order members was different for each treatment.
Microorganism is known to play a useful role in various fields of agriculture. Especially, it was used to promote growth and development of plants such as nitrogen fixation, nutrients supply, hormone production, and organic material degradation, and applied as one of solutions of eco-friendly agricultural management for sustainable crop production. We then isolated about 500 strains to secure available bacteria by serial dilution method on R2A agar plate from arable soils such as paddy soil, orchard soil and green house soil. These strains were identified on the basis of 16S rRNA gene sequence and assayed metabolic ability on 12 carbon sources and activities of lipase, protease, chitinase, cellulase and phosphate solubilization. We listed the inventory on biological activity of these isolates. In soil under long term fertilizer practices, the soil treated with 3 components (nitrogen, phosphate, potassium) has higher microbial activity than the soil treated no fertilizer. We considered that the use of soil conditioner gives some additive effects to soil microbial activity because it was showed that the soil treated with 3 components has lower microbial activity than the soil treated with silicate, lime and compost. With eleven soil samples (6 organic and 5 conventional farming soils) collected from 10 pepper-cultivating plastic film houses, the average well color development (AWCD) was measured for 6 days using ECO plate to estimate carbon substrate utilization of bacterial communities. It was observed that organic farming soils had more diverse bacterial genera than conventional farming soils. The AWCDs of organic farming soils were higher than those of conventional farming soil. It was also revealed that bacterial communities of organic farming soils had higher carbon utilization rate than those of conventional farming soils.
This study indicates that organic farming soils have more diverse bacterial community and higher metabolic potential than conventional farming soils. Currently, we are carrying out pyrosequencing, which can analyze diversity of unculturable bacteria for overcoming drawback of culturable method.
This study was conducted to aim at investigating laws and legal systems related to bio-pesticides management and grasping bio-pesticide market scale. Global market scale of bio-pesticides is increasing by about 60% every year, and by 2010, will be estimated to increase from 672 to 1,075 Million US$. The other side, Global market scale of chemical pesticides is about 26 billion US$ and reducing by 7.2% every year. Of chemical pesticides, the portion of herbicides, insecticides and fungicides is 44%, 28% and 19%, respectively. In 2010, global market scale of pesticides will be estimated to be 25billion US$ and of total of the pesticide market, bio-pesticides will estimated to be occupied 4.25%. In other hand, in 2008, production volume and marketing money of bio-pesticides was 49M/T and 1,780million won(0.15% of total pesticide market scale) in Korea, respectively. However, actual market scale of bio-pesticides will be estimated to be 82billion won in 2010.

목차 Contents

• 제출문 ... 1
• 요약문 ... 2
• SUMMARY ... 6
• 제1장 서 론 ... 9
• 제2장 국내외 기술개발 현황 ... 11
• 제3장 연구개발수행 내용 및 결과 ... 14
• 제1절 유용미생물의 효능평가 및 현장활용 증진기슬 개발 ... 14
• 제2절 다기능 미생물의 농가실용화기술 개발 ... 18
• 제3절 미생물 비료자원개발을 위한 농경지 미생물의 다양성 평가 ... 24
• 제4절 선발 미생물 자원의 기능평가 및 인벤토리 구축 ... 30
• 제5절 농업용 미생물 이용 유기농가의 경영성과분석 ... 37
• 제4장 연구개발목표 달성도 및 대외기여도 ... 40
• 제1절 목표대비 달성도 ... 40
• 제2절 정량적 성과 ... 41
• 제5장 연구개발결과의 활용계획 ... 45
• 제6장 중요 변동사항 ... 46
• 제7장 참고문헌 ... 47
• 주요 결과 요약서 ... 60