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玉米黏液对反硝化群落多样性和活性的影响。

Influence of maize mucilage on the diversity and activity of the denitrifying community.

作者信息

Mounier E, Hallet S, Chèneby D, Benizri E, Gruet Y, Nguyen C, Piutti S, Robin C, Slezack-Deschaumes S, Martin-Laurent F, Germon J C, Philippot L

机构信息

UMR INRA 1229 Microbiologie et Géochimie des Sols, 17, rue Sully, B. V. 86510, 21065 Dijon Cedex France.

出版信息

Environ Microbiol. 2004 Mar;6(3):301-12. doi: 10.1111/j.1462-2920.2004.00571.x.

Abstract

In order to understand the effect of the maize rhizosphere on denitrification, the diversity and the activity of the denitrifying community were studied in soil amended with maize mucilage. Diversity of the denitrifying community was investigated by polymerase chain reaction (PCR) amplification of total community DNA extracted from soils using gene fragments, encoding the nitrate reductase (narG) and the nitrous oxide reductase (nosZ), as molecular markers. To assess the underlying diversity, PCR products were cloned and 10 gene libraries were obtained for each targeted gene. Libraries containing 738 and 713 narG and nosZ clones, respectively, were screened by restriction fragment analysis, and grouped based on their RFLP (restriction fragment length polymorphism) patterns. In all, 117 and 171 different clone families have been identified for narG and nosZ and representatives of RFLP families containing at least two clones were sequenced. Rarefaction curves of both genes did not reach a clear saturation, indicating that analysis of an increasing number of clones would have revealed further diversity. Recovered NarG sequences were related to NarG from Actinomycetales and from Proteobacteria but most of them are not related to NarG from known bacteria. In contrast, most of the NosZ sequences were related to NosZ from alpha, beta, and gammaProteobacteria. Denitrifying activity was monitored by incubating the control and amended soils anaerobically in presence of acetylene. The N2O production rates revealed denitrifying activity to be greater in amended soil than in control soil. Altogether, our results revealed that mucilage addition to the soil results in a strong impact on the activity of the denitrifying community and minor changes on its diversity.

摘要

为了解玉米根际对反硝化作用的影响,研究了添加玉米黏液的土壤中反硝化群落的多样性和活性。利用编码硝酸还原酶(narG)和一氧化二氮还原酶(nosZ)的基因片段作为分子标记,通过聚合酶链反应(PCR)扩增从土壤中提取的总群落DNA,研究反硝化群落的多样性。为评估潜在的多样性,对PCR产物进行克隆,针对每个目标基因获得了10个基因文库。分别包含738个和713个narG和nosZ克隆的文库通过限制性片段分析进行筛选,并根据其限制性片段长度多态性(RFLP)模式进行分组。总共为narG和nosZ鉴定出117个和171个不同的克隆家族,并对至少包含两个克隆的RFLP家族的代表进行了测序。两个基因的稀释曲线均未达到明显的饱和状态,这表明分析越来越多的克隆将揭示出更多的多样性。回收的NarG序列与放线菌纲和变形菌纲的NarG相关,但其中大多数与已知细菌的NarG无关。相比之下,大多数NosZ序列与α、β和γ变形菌纲的NosZ相关。通过在乙炔存在下对对照土壤和添加黏液的土壤进行厌氧培养来监测反硝化活性。一氧化二氮的产生速率表明,添加黏液的土壤中的反硝化活性高于对照土壤。总之,我们的结果表明,向土壤中添加黏液会对反硝化群落的活性产生强烈影响,而对其多样性的影响较小。

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