Monitoring molecular fingerprinting of microbial community dynamics by denaturing gradient gel electrophoresis(DGGE) and terminal-restriction fragment length polymorphism (T-RFLP)
| Autor: | Huei-Ming Lin, 林徽鳴 |
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| Rok vydání: | 2005 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 93 Soil bacteria are essential components of the biotic community in natural forests and they are largely responsible for ecosystem function, and participate in the elements circulation. Although the main diversity of life has been proven to be microbial, the vast majority of soil bacteria still remain unknown for less than 1% of environmental microorganisms can be cultured. In this study, cloning method, denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP) techniques were used to compare the prevalent microbial populations in soil and on litterfall of Nan-Jen Shan samples. Phylogenetic analysis based on PCR-amplified 16S rDNA revealed an effective tool to establish the microbiota development in soil and litterfall. The results from clone library showed that Acidobacteria were the dominants in soil samples (62.8%) and Gammaproteobacteria were the dominants in litterfall samples (51.4%). DGGE patterns also revealed that the microbial community of soil and litterfall were not the same. Seasonal changes in the structure of microbial community were significant in DGGE analysis. However, the DGGE patterns of dominant bacteria did not change in DGGE analysis no matter in soil and litterfall samples. Terminal-restriction fragment length polymorphism (T-RFLP) analysis is also used in this study for rapid comparison of the complex bacterial communities. By T-RFLP analysis of soil and litterfall samples, we got eight groups Terminal-restriction fragments (T-RFs) in soil samples and five groups T-RFs in litterfall samples. T-RFLP analysis also revealed that Acidobacteria were the dominant bacteria in soil and were not affected by season. However, the results of T-RFLP showed that Betaproteobacteria are the dominant in litterfall. Otherwise, the dominant bacteria were changing with season on litterfall. Cultivation-approached method was also applied in isolation of cellulose degrading bacteria. Five strains were isolated from soil samples, all of them can use α-cellulose as carbon source. It showed that the soil bacteria in Nan-Jen Shan might play an important role in litter decomposition mechanism. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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