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福建省稻田土壤细菌群落的16S rDNA-PCR-DGGE分析
引用本文:李友发,宋 兵,宋亚娜,郑斯平,翟焕趁,郑伟文.福建省稻田土壤细菌群落的16S rDNA-PCR-DGGE分析[J].微生物学通报,2008,35(11):1715-1720.
作者姓名:李友发  宋 兵  宋亚娜  郑斯平  翟焕趁  郑伟文
作者单位:1. 福建省农业科学院生物技术研究所,福州,350003;福建农林大学生命科学院,福州,350002
2. 福建省农业科学院生物技术研究所,福州,350003
基金项目:福建省科技计划项目,国家配套项且
摘    要:用不依赖细菌培养的16S rDNA-PCR-DGGE方法对福建省6个不同地区12个取样点的稻田土壤进行细菌群落结构分析.对12份样品直接提取其总DNA,用F341GC/R534引物扩增16SrDNA基因的V3可变区,结合DGGE(denaturing gradient gel electrophoresis)技术分析样品细菌群落组成.结果表明,福建省不同地区的稻田土壤之间细菌群落结构存在较大差异.犬体上可分为闽东、闽南、闽北、闽西4个大类.同一地区的根际土和表土样品之间也存在差异,但差异相对较低,其中龙岩根际土和表土细菌群落结构相似性最大,永泰差异性最大.回收了DGGE图谱中11个条带,测序结果经过Blast比对表明其中10个条带代表的细菌是不可培养的,显示了DGGE技术的优越性.

关 键 词:稻田土壤  细菌群落

Analysis of the Bacterial Communities in Paddy Soils in Fujian Province Using 16S rDNA -PCR- DGGE
LI You-F,SONG Bing,SONG Ya-N,ZHENG Si-Ping,ZHAI Huan-Chen and ZHENG Wei-Wen.Analysis of the Bacterial Communities in Paddy Soils in Fujian Province Using 16S rDNA -PCR- DGGE[J].Microbiology,2008,35(11):1715-1720.
Authors:LI You-F  SONG Bing  SONG Ya-N  ZHENG Si-Ping  ZHAI Huan-Chen and ZHENG Wei-Wen
Institution:(1. Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003) (2. College of Life Science, Fujian Agriculture and Forest University, Fuzhou 350002);(1. Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003) (2. College of Life Science, Fujian Agriculture and Forest University, Fuzhou 350002);Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003;Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003;(1. Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003) (2. College of Life Science, Fujian Agriculture and Forest University, Fuzhou 350002);Biotech Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003
Abstract:16S rDNA-PCR-DGGE, a cultivation-independent approach, was used to analyze the bacterial communities in paddy soils in Fujian Province. The bulk soils and rhizosphric soils were sampled from six different ecological soil regions. Total DNA was directly extracted and amplified with the F341GC and R534 primers targeting the 16S rDNA V3. The amplified fragments were analyzed by perpendicular DGGE. Cluster analysis revealed that there was a high diversity of bacterial community compositions among different soil samples tested. Basically they could be grouped to Mindong (East), Minnan (South), Minbei (North) and Minxi (West) ecological regions. 16S rDNA-PCR-DGGE profiles from bulk and rhizospheric soil in the same region showed less bacterial diversity but more similarity. The bacterial community from bulk soil and rhizospheric soil in Longyan shared the most similar DGGE banding patterns, and the banding patterns in Yongtai exhibited the highest diversity. Eleven DGGE bands recovered were re-amplified, sequenced and aligned with Blast. The results indicated that ten of the fragments belong to uncultivated bacteria implying DGGE technique having priority in analyzing uncultivated bacteria in the paddy soils.
Keywords:16S  rDNA-PCR-DGGE
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