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
【目的】基于水稻田间定位试验,利用nirS功能基因研究不同氮肥水平对稻田土壤反硝化细菌群落多样性的影响。【方法】运用PCR-DGGE(聚合酶链反应-变性梯度凝胶电泳)结合DNA克隆测序和荧光定量PCR(Real-time PCR)技术对反硝化细菌nirS基...【目的】基于水稻田间定位试验,利用nirS功能基因研究不同氮肥水平对稻田土壤反硝化细菌群落多样性的影响。【方法】运用PCR-DGGE(聚合酶链反应-变性梯度凝胶电泳)结合DNA克隆测序和荧光定量PCR(Real-time PCR)技术对反硝化细菌nirS基因进行检测,分析田间定位试验第4年不同氮肥水平下稻田nirS型反硝化细菌群落结构和丰度的变化。【结果】依据DGGE图谱计算的群落多样性指数显示,与不施肥对照处理(CK)比较,施用氮肥处理(N1:75 kg N.hm-2,N2:150 kg N.hm-2和N3:225 kg N.hm-2)可促进稻田土壤nirS型反硝化细菌群落多样性指数提高,尤其在水稻生长的齐穗期和成熟期后者均显著高于前者(P<0.05)。但群落多样性指数在N1、N2和N3处理间的差异主要表现在水稻分蘖期和齐穗期的表层土壤中,N3可显著高于N1(P<0.05)。冗余分析结果显示水稻生育时期对稻田土壤nirS型反硝化细菌群落结构的影响较大,表层和根层土壤的群落结构都与生育时期存在显著相关性(P=0.002,0.002);而不同氮肥水平对群落结构的显著性影响仅表现在稻田表层土壤中(P=0.002)。荧光定量PCR结果显示氮肥水平提高可促进稻田土壤nirS型反硝化细菌丰度增加,在水稻分蘖期和齐穗期内表层和根层土壤的nirS基因拷贝数均存在CK<N1<N2<N3的趋势,且以齐穗期时在表层土壤中的差异最大,不同处理间的差异都达到显著水平(P<0.05)。同时,本研究获得稻田反硝化细菌nirS基因片段序列8条登录GenBank(登录号:JX997923、JX997924、JX997926—JX997931)。此外,本试验中N1、N2和N3处理比CK处理分别增产59%、92%和107%,表层和根层土壤中NO3--N含量也随氮肥用量提高而增加。【结论】氮肥用量增加促进了稻田土壤nirS型反硝化细菌丰度及群落多样性指数的提高,尤其在稻田表层土壤中。氮肥水平提高还可改变稻田表层土壤nirS型反硝化细菌的群落结构。综合分�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).
文摘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.
文摘【目的】基于水稻田间定位试验,利用nirS功能基因研究不同氮肥水平对稻田土壤反硝化细菌群落多样性的影响。【方法】运用PCR-DGGE(聚合酶链反应-变性梯度凝胶电泳)结合DNA克隆测序和荧光定量PCR(Real-time PCR)技术对反硝化细菌nirS基因进行检测,分析田间定位试验第4年不同氮肥水平下稻田nirS型反硝化细菌群落结构和丰度的变化。【结果】依据DGGE图谱计算的群落多样性指数显示,与不施肥对照处理(CK)比较,施用氮肥处理(N1:75 kg N.hm-2,N2:150 kg N.hm-2和N3:225 kg N.hm-2)可促进稻田土壤nirS型反硝化细菌群落多样性指数提高,尤其在水稻生长的齐穗期和成熟期后者均显著高于前者(P<0.05)。但群落多样性指数在N1、N2和N3处理间的差异主要表现在水稻分蘖期和齐穗期的表层土壤中,N3可显著高于N1(P<0.05)。冗余分析结果显示水稻生育时期对稻田土壤nirS型反硝化细菌群落结构的影响较大,表层和根层土壤的群落结构都与生育时期存在显著相关性(P=0.002,0.002);而不同氮肥水平对群落结构的显著性影响仅表现在稻田表层土壤中(P=0.002)。荧光定量PCR结果显示氮肥水平提高可促进稻田土壤nirS型反硝化细菌丰度增加,在水稻分蘖期和齐穗期内表层和根层土壤的nirS基因拷贝数均存在CK<N1<N2<N3的趋势,且以齐穗期时在表层土壤中的差异最大,不同处理间的差异都达到显著水平(P<0.05)。同时,本研究获得稻田反硝化细菌nirS基因片段序列8条登录GenBank(登录号:JX997923、JX997924、JX997926—JX997931)。此外,本试验中N1、N2和N3处理比CK处理分别增产59%、92%和107%,表层和根层土壤中NO3--N含量也随氮肥用量提高而增加。【结论】氮肥用量增加促进了稻田土壤nirS型反硝化细菌丰度及群落多样性指数的提高,尤其在稻田表层土壤中。氮肥水平提高还可改变稻田表层土壤nirS型反硝化细菌的群落结构。综合分�
