Coelho M R R, Carneiro N P, Marriel I E, Seldin L
Universidade Federal do Rio de Janeiro, Instituto de Microbiologia Prof. Paulo de Góes, Rio de Janeiro, RJ, Brazil.
Lett Appl Microbiol. 2009 May;48(5):611-7. doi: 10.1111/j.1472-765X.2009.02578.x. Epub 2009 Mar 9.
To develop a polymerase chain reaction (PCR)-based approach for the detection of nifH gene-containing Paenibacillus in environmental samples.
The primers, nifHPAENf and nifHPAENr, were designed and tested with DNA from: (i) strains of different nitrogen-fixing Paenibacillus species, (ii) strains of other nitrogen-fixing genera and (iii) rhizosphere of sorghum sown in Cerrado soil amended with either 12 or 120 kg ha(-1) of nitrogen fertilizer. All nitrogen-fixing Paenibacillus strains tested and the DNA samples from rhizosphere soil were amplified when these primers were used, generating a 280 bp fragment. When the PCR products obtained from both sorghum rhizospheres were cloned and sequenced, the majority of the clones analysed could be identified as Paenibacillus durus. Moreover, a greater diversity in the nifH sequences could be observed in the rhizosphere treated with a high amount of nitrogen fertilizer.
Nitrogen fertilization slightly influenced the structure of the nifH gene-containing Paenibacillus community in sorghum rhizospheres cultivated in Cerrado soil.
The PCR detection method developed is adequate to assess the presence of nifH gene-containing Paenibacillus in the environment and can be used in future to determine the ecological role of this group of micro-organisms for the nitrogen input to the plants.
开发一种基于聚合酶链反应（PCR）的方法，用于检测环境样品中含nifH基因的芽孢杆菌属。
设计引物nifHPAENf和nifHPAENr，并用于以下DNA的检测：（i）不同固氮芽孢杆菌属菌株；（ii）其他固氮属菌株；（iii）在施用12或120 kg ha⁻¹氮肥改良的塞拉多土壤中播种的高粱根际土壤。使用这些引物时，所有测试的固氮芽孢杆菌属菌株以及根际土壤DNA样本均被扩增，产生一个280 bp的片段。对从两个高粱根际获得的PCR产物进行克隆和测序后，分析的大多数克隆可鉴定为硬芽孢杆菌。此外，在施用大量氮肥的根际中，可观察到nifH序列具有更大的多样性。
氮肥对塞拉多土壤中种植的高粱根际含nifH基因的芽孢杆菌属群落结构有轻微影响。
所开发的PCR检测方法足以评估环境中含nifH基因的芽孢杆菌属的存在，未来可用于确定这组微生物对植物氮输入的生态作用。
