【作者】 辜运富； 张小平； 涂仕华； Kristina Lindstrm；

【Author】 GU Yunfu~1,ZHANG Xiaoping1,TU Shihua~2,Kristina Lindstr(o|¨)m~3 (1 College of Resource and Environment,Sichuan Agricultural University,Yaan 625014) (2 Soil and Fertilizer Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066) (3 Department of Applied Chemistry and Microbiology,University of Helsinki,Helsinki 00014)

【机构】 四川农业大学资源与环境学院微生物学系； 四川省农业科学院土壤肥料研究所； 芬兰赫尔辛基大学应用化学与微生物学系；

【摘要】 以四川省农科院于1982年在四川遂宁市船山区的石灰性紫色土上建立的"NPK长期肥料定位试验"为研究对象,通过16S rDNA的限制性酶切片段长度多态性(16S rDNA-RFLP)和变性梯度凝胶电泳(DGGE)技术研究了长期不同施肥制度下的古菌群落结构特点。结果表明,长期定位施肥对土壤中的古菌形成明显的影响,具有不同的DGGE图谱。NP,NM和NPKM长期肥料处理下的土壤古菌多样性指数低于M、NPM、CK、N和NPK肥料处理。在DGGE图谱的基础上,选择种植水稻和小麦的NPK处理土壤DNA为样品,对土壤的古菌进行16S rDNA酶切、克隆、测序分析。系统发育分析发现,石灰性紫色土中的古菌具有明显的多样性,与泉古菌界的古菌明显不同。所有古菌克隆子聚在3个群里,这些古菌均与不同陆生环境中的古菌聚在一起。对DGGE图谱的聚类分析发现,不管是石灰性紫色土种植水稻还是小麦,8种施肥处理都聚在3个群里。种植水稻时,M和NPM肥料处理下的土壤古菌聚成第一个群,NP处理下的聚成第二个群,另外5种施肥处理(包括NPKM,NM,CK,N和NPM)聚成第三个群。种植小麦时,NPKM和M处理下的土壤古茵聚成一个群,NP处理下的聚成第二个群,N,NPK,NM,NPM和CK处理下的聚成第三个群。显示不同作物种植对土壤古菌也有一定影响。更多还原

【Abstract】 Based on the ’N,P,K long-term fertilization field experiment’ which was established by the Soil and Fertilizer Institute of Sichuan Academy of Agricultural Sciences on the Calcareous Purplish Soil in 1982 located in Chuanshan district,Suining city,Sichuan province,China.the influence of M,MN,MNP,MNPK,CK,N,NP and NPK fertilizer treatments on soil archaea community structure were analyzed by using the 16S rDNA restriction fragment length polymorphism(16S rDNA-RFLP) method and denaturing gradient gel electrophoresis(DGGE).The results showed long term fertilizer could make great impact on the soil archaea community. Denaturing gradient gel electrophoresis(DGGE) analysis revealed that wheat and rice cropping had different impact on the soil archaea community structure,the richness and diversity of archaea community in the soil under NM,NP and NPKM were lower than those under the other fertilizer treatments(as M,NPM,CK,N,and NPK).Phylogenetic and nucleotide signature analyses of these cloned small subunit ribosomal RNA gene sequences revealed 3 cluster of Archaea from a soil microbial community that diverge deeply from the crenarchaeotal line of descent and has the closest affiliation to the lineage of terrestrial ecosystem Archaea.Cluster analysis of the DGGE profiles showed that archaea communities under 8 fertilizer treatments with rice and wheat cultivation were all clustered into 3 groups.In the soil with rice cultivation,the archaea communities in soil amended with M and NPM were in the same cluster.Archaea communities exposed to NP fell into the second group,while those exposed to the NPKM,NM,CK,N and NPM treatments were in the third group. In the soil with wheat cultivation,archaea community under NP was clustered into a single cluster. Those under NPKM and M were fallen into the second cluster,while those in N,NPK,NM,NPM and CK fertilizer treatments were in the third cluster.更多还原