scholarly journals Method optimization for denaturing gradient gel electrophoresis (DGGE) analysis of microflora from Eucalyptus sp. wood chips intended for pulping

2014 ◽  
Vol 13 (3) ◽  
pp. 356-365 ◽  
Author(s):  
Ramnath Lucretia ◽  
Bush Tamara ◽  
Govinden Roshini
2005 ◽  
Vol 71 (2) ◽  
pp. 993-1003 ◽  
Author(s):  
M. Bergsma-Vlami ◽  
M. E. Prins ◽  
M. Staats ◽  
J. M. Raaijmakers

ABSTRACT The genotypic diversity of antibiotic-producing Pseudomonas spp. provides an enormous resource for identifying strains that are highly rhizosphere competent and superior for biological control of plant diseases. In this study, a simple and rapid method was developed to determine the presence and genotypic diversity of 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas strains in rhizosphere samples. Denaturing gradient gel electrophoresis (DGGE) of 350-bp fragments of phlD, a key gene involved in DAPG biosynthesis, allowed discrimination between genotypically different phlD + reference strains and indigenous isolates. DGGE analysis of the phlD fragments provided a level of discrimination between phlD + genotypes that was higher than the level obtained by currently used techniques and enabled detection of specific phlD + genotypes directly in rhizosphere samples with a detection limit of approximately 5 × 103 CFU/g of root. DGGE also allowed simultaneous detection of multiple phlD + genotypes present in mixtures in rhizosphere samples. DGGE analysis of 184 indigenous phlD + isolates obtained from the rhizospheres of wheat, sugar beet, and potato plants resulted in the identification of seven phlD + genotypes, five of which were not described previously based on sequence and phylogenetic analyses. Subsequent bioassays demonstrated that eight genotypically different phlD + genotypes differed substantially in the ability to colonize the rhizosphere of sugar beet seedlings. Collectively, these results demonstrated that DGGE analysis of the phlD gene allows identification of new genotypic groups of specific antibiotic-producing Pseudomonas with different abilities to colonize the rhizosphere of sugar beet seedlings.


2006 ◽  
Vol 6 (2) ◽  
pp. 153-159 ◽  
Author(s):  
L. Ho ◽  
D. Hoefel ◽  
W. Aunkofer ◽  
T. Meyn ◽  
A. Keegan ◽  
...  

Biological sand filters were assessed for their ability to remove geosmin, 2-methylisoborneol (MIB) and microcystin-LR. Microcystin-LR was the most readily degradable metabolite with a maximum lag period of only 5 days before it was undetected in the filter effluent. Geosmin and MIB were difficult to degrade, with a period in excess of 75 days before greater than 95% removal was achieved. A microcystin-degrading gene was detected in the biofilm from one of the filters, confirming that the biofilm possessed the ability to degrade microcystin. A Sphingomonas sp. was identified as a potential geosmin degrader based on denaturing gradient gel electrophoresis (DGGE) analysis. DGGE analysis revealed a more complex bacterial community during the degradation of MIB, suggesting that more than one bacterium may be responsible for its degradation.


2020 ◽  
Vol 51 (2) ◽  
pp. 125-146
Author(s):  
Nasiruddin Nasiruddin ◽  
Yu Zhangxin ◽  
Ting Zhao Chen Guangying ◽  
Minghui Ji

We grew cucumber in pots in greenhouse for 9-successive cropping cycles and analyzed the rhizosphere Pseudomonas spp. community structure and abundance by PCR-denaturing gradient gel electrophoresis and quantitative PCR. Results showed that continuous monocropping changed the cucumber rhizosphere Pseudomonas spp. community. The number of DGGE bands, Shannon-Wiener index and Evenness index decreased during the 3rd cropping and thereafter, increased up to the 7th cropping, however, however, afterwards they decreased again. The abundance of Pseudomonas spp. increased up to the 5th successive cropping and then decreased gradually. These findings indicated that the structure and abundance of Pseudomonas spp. community changed with long-term cucumber monocropping, which might be linked to soil sickness caused by its continuous monocropping.


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