摘要：本研究对来自黑龙江伊春凉水、黑龙江帽儿山、吉林长白山、山东博山、山东泰山、福建、广东、江西以及云南等不同地区、不同植被的森林土壤进行DNA的提取和纯化。对上述不同来源的森林土壤中细菌和古菌的16S rDNA进行PCR扩增和PCR产物的半定量，比较土壤中细菌和古菌的相对丰度。结果表明，由于森林土壤具有pH值较低、腐殖酸含量较高的特点，无论是Griffths法还是试剂盒方法的提取都会有较多腐殖酸残留在DNA提取液中，这会干扰进一步的分子分析；纯化后的DNA进行PCR扩增效果较好。通过PCR产物的半定量结果我们可以初步判断森林土壤中细菌丰度高于古菌的丰度。
关键词：森林土壤， DNA提取，16S rDNA，细菌，古菌
Abstract:In this study, we extracted and purified DNA of forest soils sampled from different area and under different vegetation types of Liangshui Nature Reserve in Heilongjiang Province, Maoershan in Heilongjiang Province, ChangBai Mountain in Jilin Province, Boshan in Shandong Province, Mt.Taishan in Shandong Province, and of Fujian Province, Guangdong Province, Jiangxi Province and Yunnan Province . The 16S rDNA of bacteria and archaea were amplified from above-mentioned different forest soils, and the PCR products were semi-quantitated in order to compare the relative abundance of the bacteria and archaea in the forest soils. The results showed that the humic acid whose content is high in the soil because of the low soil pH condition is hardly to be cleaned either by Griffths method or by using extraction kit. The residue of the humic acid may interfere with further molecular analysis. The purified DNA is proper to be used for PCR amplification. Initial semi-quantitative PCR results showed that the abundance of bacteria is higher than that of archeaea in forest soils.
Key words: forest soil, DNA extraction, 16S rDNA, bacteria, archaea
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