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不同施肥水平下旱地土壤细菌群落多样性的RFLP分析 被引量:9

RFLP ANALYSIS ON MICROBIAL DIVERSITY OF DRY LAND SOILS DIFFERENTIN FERTILIZATION LEVEL
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摘要 采用直接提取土壤微生物总DNA的方法,通过对不施肥(CK)、低量施肥(F1,N∶P2O5∶K2O为0·1gkg-1∶0·075gkg-1∶0·15gkg-1)、适量施肥(F2,N∶P2O5∶K2O为0·2gkg-1∶0·15gkg-1∶0·3gkg-1)和过量施肥(F3,N∶P2O5∶K2O为1·0gkg-1∶0·75gkg-1∶1·5gkg-1)等4种不同施肥水平土样DNA提取,扩增细菌16SrDNA基因片段,建立克隆文库。用限制性内切酶HhaI和RsaI进行PCR-RFLP分析,分别得到146、133、187、170个酶切类型。采用α多样性的测度对试验结果进行分析统计表明,不同处理间土壤细菌的多样性和物种丰富度均为F2>F3>CK>F1,表明合理施肥有利于土壤细菌的多样性;λ、R2、dMa和E指数在不同施肥处理间的变异系数达到12·86%~118·9%,尤其是Simpson指数λ变化最为敏感,处理间的差异最大。序列分析和系统发育树结果表明,不同施肥处理下供试土壤优势细菌的分布发生了改变。 Total genomic DNAs were extracted from soil microorganisms in four soils different in fertilization treatment with the direct extraction method:The four treatments were CK (soil without fertilization),F1 (low in fertilization level,N∶P2O5∶K2O to 0.1 g kg-1:0.075 g kg-1∶0.15 g kg-1),F2 (normal in fertilization level,N∶P2O5∶K2O to 0.2 g kg-1∶0.15 g kg-1∶0.3 g kg-1) and F3 (excessive fertilization,N∶P2O5∶K2O to 1.0 g kg-1∶0.75 g kg-1∶1.5 g kg-1) 16S rDNAs of the extracted DNA were amplified using bacterial universal primers and clone libraries set up PCR-RFLP analysis was carried out with restriction endonuclease Rsa I and HhaI,and 146,133,187 and 170 types of restriction endonucleases were obtained from these samples. Statistical analysis of the data using α diversity measurement,which,revealed that the four treatments followed an decreasing order of F2 〉 F3 〉 CK 〉 F1 in terms of bacterial diversity and species abundance,indicating that proper fertilization was conducive to bacterial diversity in soil. Variation coefficients of indices of λ,R2, dMa and E between different treatments fell in the range from 12.86 % to118.9 %,especially for Simpson index λ,which was the most sensitive,and varied the sharpest. Results of sequencing and phylogenetic analysis proved clearly that,compared with CK,the fertilization treatments demonstrated obvious changes in distribution of dominant bacteria community.
出处 《土壤学报》 CAS CSCD 北大核心 2010年第2期347-353,共7页 Acta Pedologica Sinica
基金 黄土高原土壤侵蚀与旱地农业国家重点实验室基金(10501-178)资助
关键词 施肥水平 旱地土壤 细菌多样性 RFLP 16S rDNA Fertilization level Dry land soil Bacteria diversity RFLP 16S rDNA
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