Schmalenberger Achim, Hodge Sarah, Bryant Anna, Hawkesford Malcolm J, Singh Brajesh K, Kertesz Michael A
Faculty of Life Sciences, University of Manchester, Oxford Rd, Manchester M13 9PT, UK.
Environ Microbiol. 2008 Jun;10(6):1486-500. doi: 10.1111/j.1462-2920.2007.01564.x. Epub 2008 Feb 15.
Sulfonates are a key component of the sulfur present in agricultural soils. Their mobilization as part of the soil sulfur cycle is mediated by rhizobacteria, and involves the oxidoreductase AsfA. In this study, the effect of fertilization regime on rhizosphere bacterial asfA distribution was examined at the Broadbalk long-term wheat experiment, Rothamsted, UK, which was established in 1843, and has included a sulfur-free treatment since 2001. Direct isolation of desulfonating rhizobacteria from the wheat rhizospheres led to the identification of several Variovorax and Polaromonas strains, all of which contained the asfA gene. Rhizosphere DNA was isolated from wheat rhizospheres in plots fertilized with inorganic fertilizer with and without sulfur, with farmyard manure or from unfertilized plots. Genetic profiling of 16S rRNA gene fragments [denaturing gradient gel electrophoresis (DGGE)] from the wheat rhizospheres revealed that the level of inorganic sulfate in the inorganic fertilizer was correlated with changes in the general bacterial community structure and the betaproteobacterial community structure in particular. Community analysis at the functional gene level (asfA) showed that 40% of clones in asfAB clone libraries were affiliated to the genus Variovorax. Analysis of asfAB-based terminal restriction fragment length polymorphism (T-RFLP) fingerprints showed considerable differences between sulfate-free treatments and those where sulfate was applied. The results suggest the occurrence of desulfonating bacterial communities that are specific to the fertilization regime chosen and that arylsulfonates play an important role in rhizobacterial sulfur nutrition.
磺酸盐是农业土壤中硫的关键组成部分。作为土壤硫循环的一部分,它们的迁移由根际细菌介导,并且涉及氧化还原酶AsfA。在本研究中,于英国罗斯林斯特德的Broadbalk长期小麦试验中考察了施肥制度对根际细菌asfA分布的影响,该试验于1843年设立,自2001年起包括无硫处理。从小麦根际直接分离脱硫根际细菌,鉴定出了几种贪噬菌属和极地单胞菌属菌株,它们均含有asfA基因。从施用无机肥(含硫和不含硫)、施用农家肥的小区或未施肥小区的小麦根际中分离根际DNA。对小麦根际16S rRNA基因片段进行遗传图谱分析[变性梯度凝胶电泳(DGGE)]表明,无机肥中无机硫酸盐的含量与一般细菌群落结构的变化相关,特别是与β-变形菌群落结构的变化相关。在功能基因水平(asfA)进行群落分析表明,asfAB克隆文库中40%的克隆隶属于贪噬菌属。基于asfAB的末端限制性片段长度多态性(T-RFLP)指纹图谱分析表明,无硫酸盐处理与施用硫酸盐处理之间存在显著差异。结果表明,存在特定于所选施肥制度的脱硫细菌群落,并且芳基磺酸盐在根际细菌的硫营养中起重要作用。