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TESSARO, S. Study of the methanotrophic community present in chronosequence soils from Amazon. 2012. 94 p. Masters thesis (Microbiology) Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2012. Methanotrophic bacteria are known to utilize methane as a single source of carbon and energy, and have an important role in regulating methane flux between soil and atmosphere. Methane is a major gas contributing to global warming on Earth, taking an action 25 times greater than CO2, relative to heat retention. The Amazon forest plays an important role in the global carbon cycle, having a significant carbon reservoir. Deforestation of the original forest for the conversion to pasture is a major problem faced by the Amazon forest. Changes in land use lead to changes in soil microbial community, altering the functions of this community. The objective of this study was to investigate the presence and the diversity of methanotrophic bacteria in samples of Amazon forest chronosequence soils, where there have been a conversion of forest to pasture, and then to secondary forest, and verify if such soil management influences the structure of the methanotrophic community. Enrichment and cultivation techniques, gene libraries of pmoA gene and application of the DNA-SIP technique were used to achieve this goal. From primary, secondary forest and pasture soil samples, enrichments were performed in order to evaluate the consumption of methane. Functional gene pmoA libraries werw constructed from total DNA extracted from soils in order to study the methanotrophic community. A pasture sample was used in the DNA-SIP 13 CH4 technique, resulting in three sequence analysis of 16S rRNA gene libraries. The pmoA gene libraries indicated the presence of methanotrophic bacteria from Families Methylocystaceae and Methylococcaceae, and most of the sequences were grouped with sequences related to Methylocystaceae Family and Methylocaldum. The evaluation of methane consumption showed that the methane consuming community is active for all samples. In the DNA-SIP 13 CH4 experiment, the recovery of labeled DNA occurred after 0.6 mmol of methane were consumed in the microcosm, incubation time t2. Gene libraries of rRNA 16S from total DNA of soil, revealed the presence of bacteria over several Phyla related to soil. In gene libraries constructed from labeled DNA, methane cycle microorganisms were identified such as the Methylocystaceae, and Methylococcaceae methanotrophic Families. Other groups of microorganisms like Methylophilaceae Family and Armatimonadetes Phylum (OP10) bacteria were also found, but their role in the methane cycle is not fully understood. The knowledge about the process of metabolites from the methane oxidation by these microorganisms may be uncertain, although the presence of this microorganisms in the environment may be important.

Estudo da comunidade de bactérias metanotróficas em uma cronossequência de solos da Amazônia.