Bacterial diversity in organic soil layers of subalpine and alpine forests at the end of freeze-thaw periods

With the use of a denature gradient gel electrophoresis (DGGE) method, we investigated the bacterial diversity in both organic soil (OL) and mineral soil (MS) layers at the end of freeze-thaw periods in four typical forests, i.e. a primary fir (Abies faxoniana) forest, a secondary fir forest, a 20-year-old spruce (Picea asperata) plantation and a 15-year-old spruce plantation in the subalpine and alpine regions of western Sichuan Province. The bacterial DNA of the soil was extracted after chemical breakage. High specific V3 fragments of 16S rDNA were obtained from purified DNA by a touch-down PCR technique. A large number of bands were observed on the DGGE gel, with different intensities and distances among samples, indicating high bacterial diversity of the OL in the four forests after a seasonal freeze-thaw period. Species similarity, Shannon-Wiener richness and Simpson dominance indices of bacterial communities in the OL varied greatly with soil thickness and elevation. These three indices varied only slightly in the MS. As well, low-temperature resistance, temperature sensitive and non-temperature sensitive bacterial clones were observed based on the clones and DNA sequence analyses for 10 specific bands. These results suggested that the changes in the environment driven by temperature variation had strong effects on the diversity of bacterial communities of the OL at the end of the freeze-thaw period, which provides sufficient evidence for the understanding of soil ecological processes during freeze-thaw seasons. 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