【目的】研究施用无机氮肥对小麦田土壤nirS型反硝化细菌多样性的影响。【方法】通过构建反硝化细菌亚硝酸盐还原酶nirS基因克隆文库,采用限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)技术分析了施氮肥处理和...【目的】研究施用无机氮肥对小麦田土壤nirS型反硝化细菌多样性的影响。【方法】通过构建反硝化细菌亚硝酸盐还原酶nirS基因克隆文库,采用限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)技术分析了施氮肥处理和不施氮肥处理土壤中nirS型反硝化细菌的多样性。【结果】两种处理各自分别得到了27个可操作分类单元(Operational Taxa Units,OTUs),其中有9个OTUs在两个处理中相同。虽然两种处理中nirS反硝化细菌的香农-威纳指数,Simpon指数,丰富度指数,均匀度指数相近,但是土壤的OTU类型发生了很大变化。通过对施氮肥处理nirS文库中11个代表性nirS克隆子的序列分析,有10个克隆子与数据库中的nirS序列的相似度在73%~95%之间,有1个序列在数据库中找不到相似序列。【结论】施氮肥在短期内显著改变了土壤中nirS型反硝化细菌群落结构的构成。展开更多
Denitrification is one of the major processes causing nitrogen loss from arable soils.This study aimed to investigate the responses of nir S-type denitrifier communities to different chronic fertilization regimes acro...Denitrification is one of the major processes causing nitrogen loss from arable soils.This study aimed to investigate the responses of nir S-type denitrifier communities to different chronic fertilization regimes across the black soil region of Northeast China.Soil samples were collected from sites located in the north(NB),middle(MB),and south(SB)of the black soil region of Northeast China,each with four chronic fertilization regimes:no fertilizer(No F),chemical fertilizer(CF),manure(M),and chemical fertilizer plus manure(CFM).Methods of quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing were applied to assess the abundance and composition of denitrifier communities by targeting the nir S gene.The results showed that the M and CFM regimes significantly increased the abundances of nir S-type denitrifiers compared with No F at the three locations.The majority of nir S sequences were grouped as unclassified denitrifiers,and the different fertilizers induced little variation in the relative abundance of known nir S-type denitrifier taxa.Over 90%of the sequences were shared among the four fertilization regimes at each location,but none of the abundant operational taxonomic units(OTUs)were shared among the three locations.Principal coordinate analysis(PCo A)revealed that the communities of nir S-type denitrifier were separated into three groups that corresponded with their locations.Although similar fertilization regimes did not induce consistent changes in the nir S-type denitrifier communities,soil p H and NO-3-N content simultaneously and significantly influenced the structure of nir S-type denitrifier communities at the three locations.Our results highlight that geographical separation rather than chronic fertilization was the dominant factor determining the nir S-type denitrifier community structures,and similar chronic fertilization regimes did not induce consistent shifts of nir S-type denitrifier communities in the black soils.展开更多
基金the Strategic Priority Research Program of Chinese Academy of Sciences(No.XDB15010103)the National Key Research and Development Program of China(No.2017YFD0200604)+1 种基金the National Natural Science Foundation of China(No.41771284)the Chinese Biodiversity Monitoring and Research Network(Sino BON).
摘要Denitrification is one of the major processes causing nitrogen loss from arable soils.This study aimed to investigate the responses of nir S-type denitrifier communities to different chronic fertilization regimes across the black soil region of Northeast China.Soil samples were collected from sites located in the north(NB),middle(MB),and south(SB)of the black soil region of Northeast China,each with four chronic fertilization regimes:no fertilizer(No F),chemical fertilizer(CF),manure(M),and chemical fertilizer plus manure(CFM).Methods of quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing were applied to assess the abundance and composition of denitrifier communities by targeting the nir S gene.The results showed that the M and CFM regimes significantly increased the abundances of nir S-type denitrifiers compared with No F at the three locations.The majority of nir S sequences were grouped as unclassified denitrifiers,and the different fertilizers induced little variation in the relative abundance of known nir S-type denitrifier taxa.Over 90%of the sequences were shared among the four fertilization regimes at each location,but none of the abundant operational taxonomic units(OTUs)were shared among the three locations.Principal coordinate analysis(PCo A)revealed that the communities of nir S-type denitrifier were separated into three groups that corresponded with their locations.Although similar fertilization regimes did not induce consistent changes in the nir S-type denitrifier communities,soil p H and NO-3-N content simultaneously and significantly influenced the structure of nir S-type denitrifier communities at the three locations.Our results highlight that geographical separation rather than chronic fertilization was the dominant factor determining the nir S-type denitrifier community structures,and similar chronic fertilization regimes did not induce consistent shifts of nir S-type denitrifier communities in the black soils.