好氧降解木质素化合物的海洋微生物分离与鉴定
毕业论文关键词:木质素;芳香族化合物;香草酸；海洋细菌;
Isolation and Identification of Marine Aerobic Bacteria Related to Lignin Degradation
Abstract:Lignin in the cell walls of the terrestrial vascular plants is very rich in the content of the nature , so it only exists on land, not in marine plant. However, lignin and its derivatives continuously go into the ocean along the river, and research on the degradation of lignin in soil microbial is also very in-depth. Meanwhile , research on the degradation of lignin in the marine environment is still not very clear. Therefore, the research goal of this paper is to separate and screen out bacterias that have the degradation and utilization capacity of vanillic acid from the marine environment by using lignin derivative (vanillic acid) as the sole carbon source, in order to reveal the exist of microbes that can degrade lignin related to aromatic compounds in the marine environment .Three strains are isolated from the seawater in Lianyungang and Seabed sediment by using the sole carbon source. The 16S rDNA sequencing of two strains(VA1-2 and VA1-3) found that VA1-2 has the similarity of 95.8% to Pseudoalteromonas mariniglutinosa and VA1-3 has the similarity of 95.8% to Halomonas chromatireducens M1S6-22. The optimum temperature of VA1-3 is about 35 ºC; the optimum salt concentration of VA1-3 is 22.8 g / L. Pseudoalteromonas mariniglutinosa and Halomonas chromatireducens usually grow up in marine environment. Up to now, there are hardly any reports on the degradation of aromatic compounds. The success of this research will open up a new way for the development of the marine biological resources, but also provide a new research direction to solve the problem about the lignin pollution environment.
Key words : lignin; aromatic compounds; vanillic acid ;marine bacteria
目录
引言 1
1 材料与方法 2
1.1供试材料 2
1.1.1 海水 3
1.1.2 试剂化合物 3
1.1.3 培养基 3
1.1.4 各种试剂溶液的制备 3
1.2 仪器 4
1.3方法 5
1.3.1采样 5
1.3.2 碳源配制 5
1.3.3 富集培养降解香草酸海洋微生物 5
1.3.4 传代 5
1.3.5 菌种保存 5
1.3.6 平板划线 5
1.3.7 PCR扩增 6
1.3.8 PCR反应产物检测及纯化 6
1.3.9 测序 7
1.3.10 革兰氏染色 7