DGGE and T-RFLP Analysis of Bacterial Succession during Mushroom Compost Production and Sequence-aided T-RFLP Profile of Mature Compost |
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Authors: | Anna J Székely Rita Sipos Brigitta Berta Balázs Vajna Csaba Hajdú Károly Márialigeti |
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Institution: | 1.Department of Microbiology,E?tv?s Loránd University,Budapest,Hungary;2.Strain Research and Molecular Biological Laboratory,Korona Spawn Factory,Demjén,Hungary |
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Abstract: | The amount of button mushroom (Agaricus bisporus) harvested from compost is largely affected by the microbial processes taking place during composting and the microbes inhabiting
the mature compost. In this study, the microbial changes during the stages of this specific composting process were monitored,
and the dominant bacteria of the mature compost were identified to reveal the microbiological background of the favorable
properties of the heat-treated phase II mushroom compost. 16S ribosomal deoxyribonucleic acid (rDNA)-based denaturing gradient
gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) molecular fingerprinting methods
were used to track the succession of microbial communities in summer and winter composting cycles. DNA from individual DGGE
bands were reamplified and subjected to sequence analysis. Principal component analysis of fingerprints of the composting
processes showed intensive changes in bacterial community during the 22-day procedure. Peak temperature samples grouped together
and were dominated by Thermus thermophilus. Mature compost patterns were almost identical by both methods (DGGE, T-RFLP). To get an in-depth analysis of the mature
compost bacterial community, the sequence data from cultivation of the bacteria and cloning of environmental 16S rDNA were
uniquely coupled with the output of the environmental T-RFLP fingerprints (sequence-aided T-RFLP). This method revealed the
dominance of a supposedly cellulose-degrading consortium composed of phylotypes related to Pseudoxanthomonas, Thermobifida, and Thermomonospora. |
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