보고서 정보
주관연구기관 |
동아대학교 Donga University |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2015-02 |
과제시작연도 |
2011 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201500010235 |
과제고유번호 |
1395022049 |
사업명 |
차세대바이오그린21 |
DB 구축일자 |
2015-07-11
|
DOI |
https://doi.org/10.23000/TRKO201500010235 |
초록
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Ⅳ. 연구개발결과
1. 작물 근권 미생물 군집 비교 분석: 토양 특성 파악, 토마토 품종 별 근권 미생물군집 비교 결과 군집구조의 차이 확인함, 각 품종별 근권의 우접 미생물 군 및 근권에서 밀도가 증가하는 Bacteroidetes 문의 미생물 확인
2. 작물근권 비교메타게놈 서열 해독: Hawaii 7996 과 Moneymaker 근권의 전 메타게놈을 각각 50 Gb 해독함
3. 작물근권 메타게놈 유래 유용 유전자원 확보 분석: 메타게놈 라이브러리 46,500 클론 확보, 지질대사 자원 확보, biocide 저항성
Ⅳ. 연구개발결과
1. 작물 근권 미생물 군집 비교 분석: 토양 특성 파악, 토마토 품종 별 근권 미생물군집 비교 결과 군집구조의 차이 확인함, 각 품종별 근권의 우접 미생물 군 및 근권에서 밀도가 증가하는 Bacteroidetes 문의 미생물 확인
2. 작물근권 비교메타게놈 서열 해독: Hawaii 7996 과 Moneymaker 근권의 전 메타게놈을 각각 50 Gb 해독함
3. 작물근권 메타게놈 유래 유용 유전자원 확보 분석: 메타게놈 라이브러리 46,500 클론 확보, 지질대사 자원 확보, biocide 저항성 자원 확보 분석
4. 항생활성 친환경 미생물 유용 유전자 분석: 친환경 미생물 Burkholderia pyrrocina CH-67의 항생활성 관련 유전자 집단 3종 확인
5. 식물 근권 우점 미생물 분리 및 식물관련 특성 규명: Flavobacterium 31종 및신종 미생물 분리 및 동정, 식물생육 촉진 활성 및 항진균 활성 확인함
6. 근권 미생물 군집이 풋마름병 저항성에 미치는 영향: 근권 미생물 군집 교환으로 풋마름병에 대한 품종 반응 변화 확인, 미생물 군집이 품종별 기능부여와 연관된확인
7. 작물 근권 비교메타게놈(whole metagenome) 정보 분석: 저항성 품종 근권에서 Bateroidetes 증가, 감수성 품종 근권에서 Betaproteobacteria 증가 확인
8. 전 메타게놈 염기서열의 조림 및 유전자 구성 분석: 유전자 구성 비에서 두 근권사이의 유의미한 차이를 파악하기 어려움, 유전자 구성 비율에서 일부 차이 확인
9. 전 메타게놈 분석 결과의 reference mapping 및 scaffolding: 6% 가량 mapping,미생물 별 mapping 비율은 군집구조와 유사함, scaffolding 결과의 분포 파악, 저항성 근권에서 신규 미생물 군 Formosa 종이 우점임을 확인
10. 친환경 미생물 유전체 분석: Burkholderia 2종, Bacillus 2종, Flavobacteria 5종 분석, 각 미생물로부터 작물 연관 유용 유전자 군 확보함(2차 대사산물 유전자 군,항생활성 유전자 군, 식물 생장 촉진 연관 유전자군)
Abstract
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Soil and plant rhizosphere are the important agricultural environments inhabiting an array of organisms. Various processes were achieved in soil, which include interactions between plants and microorganisms, metabolic reactions, and nutrient cycles. Soil microorganisms are responsible for the reacti
Soil and plant rhizosphere are the important agricultural environments inhabiting an array of organisms. Various processes were achieved in soil, which include interactions between plants and microorganisms, metabolic reactions, and nutrient cycles. Soil microorganisms are responsible for the reactions especially the rhizospheric bacteria existing in plant roots that are closely related to growth and health of plants. Microbial diversity is enormous in agricultural soils and plant rhizosphere, when estimated by a culture-independent approach.The microbial community in plant rhizosphere might be directly linked with plant function and agricultural productivity. Here, we investigated microbial community of tomato plant rhizosphere by metagenome analysis and compared the microbiome structure between bacterial wilt resistant plant and susceptible plant, Hawaii7996 and Moneymaker, respectively. We cultivated tomato cultivars under greenhouse condition with field soils at Dong-A Agricultural experimental station.
Microbial communities of tomato rhizosphere of both cultivars were analyzed by amplifying 16S rRNA genes from those rhizosphere soil and pyrosequencing of the amplified clones through GS-FLX 454 sequencer. Microbial community structure was analyzed using MG-RAST system. The most abundant microbial phylum was Proteobacteria both in bulk soil and in tomato rhizosphere soil.Compared to bulk soils, the proportions of the phyla such as Bacteroidetes and Firmicutes were significantly increased in tomato rhizosphere, while that of phylum Acidobacteria was decreased in tomato rhizosphere. Especially, bacterial members in class Flavobacteria were highly abundant in tomato rhizosphere. Comparison of microbial communities of tomato rhizosphere revealed that rhizosphere microbiome between Hawaii7996 and Moneymaker was distinguishable. The tomato rhizosphere showed the similar pattern of microbial diversity for two consecutive years.
Our result suggested that the unique microbial community forms in tomato rhizosphere in a cultivar specific manner. The analysis pipeline was also established for whole meatagenome analysis using massive sequencing data. The pipeline was used for comparative analysis of rhizospheric metagenomes of tomato cultivars that are resistant and susceptible to bacterial wilt. The results of metagenome analysis of the rhizospheres showed differences in microbial community structure and gene pool. The genome and metagenome analysis conducted in this study could be suggested as a model for other studies to understand host-microbes interaction or various environments in levels of massive sequence information. Since our microbial community analysis of tomato rhizosphere from our study revealed that members of phylum Bacteroidetes were abundant in tomato rhizosphere compared to bulk soil.We hypothesized that those bacteria dominating tomato rhizosphere contribute to the plant function.
A pair of specific primer targeting those bacteria in class Flavobacteria was designed and used to select bacterial isolates in a member of the class Flavobacteria by colony PCR of bacterial colonies derived from tomato plant rhizosphere. A number of bacteria were isolated and examined for phenotypes relevant to plant-beneficial interaction, such as biofilm formation, seedling growth promotion, and antimicrobial activities. We also analyzed genome sequences of plant-associated bacteria. The genome sequences were produced by sequencing platforms based on next generation sequecing technology. Genome sequences of two Burkholderia strains and five Flavobacterium strains were generated through this study. Genomes were analyzed for all five strains in Flavobacterium and revealed that they harbor genes for plant growth promotion, such as auxin biosynthesis and ethylene removal, and for secondary metabolites which may be involved in beneficial effect to plants.
Through the genome sequence analysis, the potential genes involved in plant growth promoting factors and antibiosis were constructed. In addition to microbiome analysis, we also investigated agriculturally important bacterial traits from the genome informations of plant-associated microbes whose genome was analysed in this study and from bioprospecting metagenome approach. We identified genes for antimicrobial activities from Burkholderia species and also lipolytic activity and biocide resistance from metagnome library constructed using tomato rhizosphere soil. We propose that the plant associated bacterial genomes and rhizospheric metagenome would be provided as useful information for productivity and disease suppression of plants and crops.
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