Long-term nitrogen fertilization alters microbial community structure and denitrifier abundance in the deep vadose zone

Purpose

The excessive use of nitrogen (N) fertilizer in intensive agriculture has increased nitrate leaching into groundwater, but its impacts on N transformation processes and the associated microbial communities in the deep vadose zone remain unclear.

Materials and methods

Soil samples from 0–1050 cm depth were collected from a 20-year field experiment with two N fertilization treatments: 0 (N0) and 600 kg N ha^?1 year^?1 (N600). Amplicon sequencing and quantitative PCR analyses were performed to profile the vertical distribution of soil microbial communities and denitrification genes.

Results and discussion

The soil microbial community structure and diversity were strongly influenced by soil depth and N fertilization. The 250 cm depth was identified as a threshold depth, as dramatically different microbial communities were found below and above this depth. Quantitative PCR results showed that the absolute abundance of denitrification genes decreased with increasing soil depth.

Conclusion

This study elucidated the profound effects of long-term N input on the composition and diversity of the microbial communities and the abundance of denitrifiers in the deep vadose zone. Our results provide basic information for use in mitigating nitrate leaching by enhancing microbial denitrification in deep vadose zones in intensive agricultural areas.

     

华北平原农田厚包气带及含水层反硝化细菌的分离鉴定及作用机制研究

华北平原农田由于长期过量施用氮肥,造成了土壤硝酸盐累积,导致地下水硝酸盐污染日趋严重。微生物的反硝化作用可将土壤中累积的硝酸盐或亚硝酸盐还原为气态产物,是消减厚包气带土壤累积的硝酸盐的重要途径。因此筛选高效反硝化微生物资源,对人工强化厚包气带土壤反硝化脱氮,阻控地下水硝酸盐污染具有重要作用。基于此,本研究采集位于华北平原的中国科学院栾城农业生态系统试验站长期施氮[施氮量为600kg(N)·hm~(-2)·a~(-1)]定位试验0～150m农田厚包气带及含水层土壤样品,从中筛选到62株细菌。16SrRNA基因序列分析表明这62株菌株与变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)中的9个属具有较高的同源性。根据系统发育树的结果,挑选7株亲缘关系较远的菌株进行反硝化潜势试验,结果表明,菌株L71、L13和L103具备反硝化产气能力。电镜观察结果表明,这3株菌均为无鞭毛的杆状细菌,其长度分别为1.0μm、1.5μm和1.5μm,只有L103具有运动能力。此外,菌株L103具有完全反硝化能力,且脱氮能力受到pH的影响,在本试验条件下,菌株L103的反硝化速率高达1.62～2.36g(KNO_3)·d~(-1)·L~(-1),具备实际应用潜力。本研究表明华北平原厚包气带土壤中存在完全反硝化微生物,并可为人工强化治理厚包气带土壤硝酸盐污染提供菌种资源和理论依据。