Application of DGGE to Improve Strategy for Bacteria! Population Responded Soils Amended with Inorganic Nitrogen and Phosphorus

摘要

The essential nutrient is important for bacterial cell growth, the cell will accumulate nutrient synthesizes its genetic material in the bacterial cell. Nucleic acids (DNA and RNA) are function as the genetic information of the cell. The 16S rRNA is necessary for the cell function and also related with growth rate. The influence of bacterial diversity in 16S ribosomal DNA (16S rDNA-) and reverse-transcribed 16S rRNA (16S rRNA) from difference soil that had been contaminated with diesel and the othercontaminated with lubricant. Both soils were amended with inorganic fertilizer (nitrogen and phosphorus) and conduced in laboratory-scale. The bacterial diversity was investigated using a culture-independent approach based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of total 16S rDNA and 16S rRNA products. The combination of rDNA and rRNA fingerprints indicated which taxa might be metabolically active in this amended soil. The twenty selected bands from both16S rDNA and 16S rRNA gel on these studies showed ten bands from 16S rDNA through to represent uncultured bacteria and other ten bands from 16S rRNA, recovered in clades containing known culture bacteria. There were relatively biggest represented in theband pattern of 16S rRNA compared with the 16S rDNA pattern, where as the 16S rRNA patterns of soil contaminated with diesel also showed five bands corresponding to a Mycobacterium-related bacterium, a Chryseobacterium-related bacterium, an Acinetobacter-related bacterium, a Pseudomonas-related bacterium, and a Bacillus-related bacterium. Additionally, the 16S rRNA patterns of soil contaminated with lubricant showed three bands with a very prominent this corresponding to a Mycobacterium-related bacterium, a Microbacterium-related bacterium and a Bacillus-related bacterium. There results suggest that the rate of RNA synthesis is highly related between the rate of growth because the results of 10 bands from 16S rRNA in both soils were related to the culturable bacteria, thus providing for the incubation periods evidence that this group of bacteria is metabolically active in soil after amended with inorganic fertilizer. It was concluded that the RNA transcript are more responsive biomarker in this study,due to the known relationship between rRNA abundance and the physiological status of known culturable bacteria.