우렁쉥이로부터 유래된 해양미생물의 이차대사산물 및 생물다양성 연구
Phylogenetic Diversity and Secondary Metabolites Production of Marine Bacteria associated with Halocynthia roretzi
- 발행기관 강릉원주대학교 대학원
- 지도교수 손영창, 권학철
- 발행년도 2010
- 학위수여년월 2010. 2
- 학위명 석사
- 학과 및 전공 일반대학원 KIST강릉분원학.연협동과정
- 실제URI http://www.dcollection.net/handler/kangnung/000000006301
- 본문언어 한국어
초록/요약 도움말
Two marine bacteria strains KME 001T and KME 002T, were isolated from the siphon tissue of a marine ascidian, Halocynthia roretzi, collected off the coast of Gangneung, Korea. KME 001T is a Gram-positive, aerobic, non-spore-forming, rod-shaped and nonmotile bacterium. Phylogenetic analysis using 16S rRNA gene sequences revealed that strain KME 001T clustered with the genus Aeromicrobium and is closely related to the species Aeromicrobium ginsengisoli, A. erythreum and A. ponti with 97.7, 97.6 and 97.5% sequence similarities, respectively. The strain is capable of growth at a variety of temperatures (10 - 42→) and over a broad pH range (5.0 ?C 10.0). NaCl was required for robust growth of the strain. The diagnostic diamino acid of the cell wall peptidoglycan is LL-diaminopimelic acid. The predominant menaquinone is MK-9(H4). The predominant fatty acids were determined to be C18:1 ┺9㎯, C16:0 and 10-methyl C18:0. The DNA-DNA hybridization analysis showed that DNA-DNA relatedness values between strain KME 001T and its nearest neighbors, A. ginsengisoli KCTC 19207T, A. erythreum KCCM 41104T and A. ponti KACC 20565T, were 49.6, 57.1 and 63.5%, respectively. The DNA G+C content of KME 001T is 75.9 mol%. Chemical investigation of the liquid culture medium of strain KME 001T led to the isolation of taurocholic acid as a major secondary metabolite. On the basis of phylogenetic and phenotypic data, bacterial strain KME 001T is classified as a new species of the genus Aeromicrobium, for which the name Aeromicrobium halocynthiae sp. nov. is proposed. The type strain is KME 001T (= JCM 15749T = KCCM 90079T). The strain KME 002T is a Gram-negative, aerobic and ovoid to rod-shaped bacterium. The phylogenetic analysis based on 16S rRNA gene sequence showed strain KME 002T fell within the radiation of family Rhodobacteraceae in the ┒-Proteobacteria and is closely related to Dinoroseobacter shibae, Roseovarious crassostreae and Pseudoruegeria aquimaris with 95.0, 94.7 and 94.5% similarities, respectively. The obligate halophilic bacterium required 1~5% (w/v) NaCl and had an absolute requirement for magnesium chloride. The cells were motile by means of a polar single flagellum and multiplied by budding. The predominant cellular fatty acid of the isolate was C18:1 ┺7㎯ and it contained Q-10 as the major ubiquinones. The DNA G+C content of the strain is 71.6 mol%. On the basis of these polyphasic characteristics of the new isolates, the bacterial strain KME 002T represents a novel genus and species of the family Rhodobacteraceae, for which the name Hasllabacter halocynthiae gen. nov., sp. nov. is proposed. The type strain is KME 002T (= JCM 16214T = KCCM 90082T).
more목차 도움말
Summary ∃
1. Introduction
1.1 Bacterial classification 1
1.2 History & Characteristics of Genus Aeromicrobium 4
1.3 History & Characteristics of Family Rhodobacteraceae 5
2. Materials and Methods
2.1 Sample Collection 7
2.2 Isolation of Microorganisms 7
2.3 Phenotypic analysis 8
2.3.1 The growth ability on various agar media 8
2.3.2 Colony and Cell morphology (Shape, Size, flagella and Motility) 8
2.3.3 Temperature, Salinity and pH test 8
2.3.4 Gram staining, Spore staining, Oxidase and Catalase test 9
2.3.5 Biophenotypic characterization 9
2.4. Chemotaxonomy 9
2.4.1 Fatty acid methyl ester and Polar lipid components 9
2.4.2 Diaminopimelic acid and Respiratory isoprenoid quinone components 10
2.5 Molecular taxonomy 10
2.5.1 DNA extraction and 16S rRNA gene sequencing 10
2.5.2 Phylogenetic analysis 11
2.5.3 GC ratio and DNA-DNA hybridization 11
3. Results and Discussion
3.1 Isolation of microorganism from the marine ascidian 13
3.2 Strain KME 001T (Aeromicrobium halocynthiae sp. nov.) 14
3.2.1 Phenotypic analysis 14
3.2.2 Chemotaxonomy 17
3.2.3 Molecular taxonomy 17
3.3 Strain KME 002T (Hasllabacter halocynthiae gen. nov., sp. nov.) 20
3.3.1 Phenotypic analysis 20
3.3.2 Chemotaxonomy 23
3.3.3 Molecular taxonomy 23
4. Description
4.1 Description of Aeromicrobium halocynthiae sp. nov. 43
4.2 Description of Hasllabacter gen. nov. 44
4.2.1 Description of Hasllabacter halocynthiae sp. nov. 45
5. References 47
