冻融末期亚高山/高山森林土壤有机层细菌多样性
Bacterial diversity in organic soil layers of subalpine and alpine forests at the end of freeze-thaw periods
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摘要: 为了解季节性融冻末期亚高山/高山森林生态系统土壤有机层细菌多样性特征,应用DGGE方法研究了川西亚高山/高山地区4个典型森林群落(冷杉原始林、冷杉次生林、20年生粗枝云杉人工林、15年生粗枝云杉人工林)土壤冻结末期土壤有机层(OL)和矿质土壤层(MS)的细菌多样性。采用化学裂解法提取土壤有机层和矿质土壤层样品总DNA获得了很好的效果,纯化后的总DNA经降落式PCR扩增得到高特异性16S rDNA V3区片段。样品PCR扩增产物经DGGE分离得到大量条带,不同样品间分离所得条带的强度和位置均有差异,表明研究样地在冻融末期具有一定的细菌群落多样性。并且,OL细菌群落结构的相似性、Shannon-Wiener物种丰富度指数和Simpson物种优势度指数随其土壤层次和森林类型的变化而变化,表明冻融末期强烈的环境变化极大地影响了土壤细菌群落结构。另外,将研究地4个样点土壤有机层细菌10个特有条带进行克隆和序列测定,结果显示了耐低温、温度敏感和非敏感优势细菌群落。这些结果表明: 冻融末期温度驱动的环境变化深刻影响了亚高山/高山森林OL细菌群落多样性。Abstract: 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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Keywords:
- subalpine-alpine forest /
- organic soil layer /
- bacterial diversity /
- DGGE
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