Campbell James H, Clark Jeb S, Zak John C
Department of Biological Sciences, Texas Tech University, P.O. Box 43131, Lubbock, TX 79409-3131, USA.
Microb Ecol. 2009 Feb;57(2):261-6. doi: 10.1007/s00248-008-9479-3. Epub 2008 Dec 24.
The polymerase chain reaction coupled with denaturing gradient gel electrophoresis (PCR-DGGE) has been used widely to determine species richness and structure of microbial communities in a variety of environments. Researchers commonly archive soil samples after routine chemical or microbial analyses, and applying PCR-DGGE technology to these historical samples offers evaluation of long-term patterns in bacterial species richness and community structure that was not available with previous technology. However, use of PCR-DGGE to analyze microbial communities of archived soils has been largely unexplored. To evaluate the stability of DGGE patterns in archived soils in comparison with fresh soils, fresh and archived soils from five sites along an elevational gradient in the Chihuahuan Desert were compared using PCR-DGGE of 16S rDNA. DNA from all archived samples was extracted reliably, but DNA in archived soils collected from a closed-canopy oak forest site could not be amplified. DNA extraction yields were lower for most archived soils, but minimal changes in bacterial species richness and structure due to archiving were noted in bacterial community profiles from four sites. Use of archived soils to determine long-term changes in bacterial community structure via PCR-DGGE appears to be a viable option for addressing microbial community dynamics for particular ecosystems or landscapes.
聚合酶链反应结合变性梯度凝胶电泳（PCR-DGGE）已被广泛用于确定各种环境中微生物群落的物种丰富度和结构。研究人员通常在常规化学或微生物分析后存档土壤样本，将PCR-DGGE技术应用于这些历史样本可评估细菌物种丰富度和群落结构的长期模式，而这是以前的技术无法实现的。然而，利用PCR-DGGE分析存档土壤的微生物群落这一领域在很大程度上尚未得到探索。为了评估存档土壤与新鲜土壤中DGGE图谱的稳定性，我们利用16S rDNA的PCR-DGGE技术，对奇瓦瓦沙漠沿海拔梯度的五个地点的新鲜土壤和存档土壤进行了比较。所有存档样本的DNA都能可靠提取，但从一个封闭树冠橡树林地点采集的存档土壤中的DNA无法扩增。大多数存档土壤的DNA提取产量较低，但在四个地点的细菌群落图谱中，因存档导致的细菌物种丰富度和结构变化极小。通过PCR-DGGE利用存档土壤确定细菌群落结构的长期变化，似乎是解决特定生态系统或景观中微生物群落动态问题的一个可行选择。
