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铜污染土壤线虫多样性的PCR-DGGE分析
作者姓名:WANG Shi-Bin  LI Qi  LIANG Wen-Ju  JIANG Yong  JIANG Si-Wei
作者单位:Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn
摘    要:A wheat pot experiment was conducted under greenhouse conditions to assess the effect of copper contamination on soil nematode diversity by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method and morphological analysis. The soil was treated with CuSO4.5H2O at the following concentrations: 0, 50, 100, 200, 400, and 800 mg kg^-1 dry soil, and the soil samples were collected at wheat jointing and ripening stages. Nematode diversity index (H′) from morphological analysis showed no difference between the control and the treated samples in either of the sampling dates. At the wheat ripening stage, nematode diversity obtained by the PCR-DGGE method decreased noticeably in the Cu800 treatment in comparison with the control. With optimization of the method of nematode DNA extraction, PCR-DGGE could give more information on nematode genera, and the intensity of the bands could reflect the abundance of nematode genera in the assemblage. The PCR-DGGE method proved promising in distinguishing nematode diversity in heavy metal coritaminated soil.

关 键 词:土壤污染  重金属污染    PCR-DGGE  线虫类

PCR-DGGE analysis of nematode diversity in Cu-contaminated soil
WANG Shi-Bin,LI Qi,LIANG Wen-Ju,JIANG Yong,JIANG Si-Wei.PCR-DGGE analysis of nematode diversity in Cu-contaminated soil[J].Pedosphere,2008,18(5):621-627.
Authors:WANG Shi-Bin  LI Qi  LIANG Wen-Ju  JIANG Yong and JIANG Si-Wei
Institution:Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn;Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China). E-mail: liq@iae.ac.cn
Abstract:Biolog,16S rRNA gene denaturing gradient gel electrophoresis (DGGE),and phospholipid fatty acid (PLFA) analyses were used to assess soil microbial community characteristics in a chronosequence of tea garden systems (8-,50-,and 90year-old tea gardens),an adjacent wasteland,and a 90-year-old forest.Biolog analysis showed that the average well color development (AWCD) of all carbon sources and the functional diversity based on the Shannon index decreased (P<0.05)in the following order:wasteland>forest>tea garden.For the DGGE analysis,the genetic diversity based on the Shannon index was significantly lower in the tea garden soils than in the wasteland.However,compared to the 90-year-old forest,the tea garden soils showed significantly higher genetic diversity.PLFA analysis showed that the ratio of Gram positive bacteria to Gram negative bacteria was significantly higher in the tea garden soils than in the wasteland,and the highest value was found in the 90-year-old forest.Both the fungal PLFA and the ratio of fungi to bacteria were significantly higher in the three tea garden soils than in the wasteland and forest,indicating that fungal PLFA was significantly affected by land-use change.Based on cluster analysis of the soil microbial community structure,all three analytical methods showed that land-use change had a greater effect on soil microbial community structure than tea garden age.
Keywords:Biolog  DGGE  microbial community structure  PLFA  tea garden soil
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