为了认识长期施肥对石灰性紫色水稻土培肥和肥力演化的作用,结合变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)和限制性酶切片段长度多态性(RFLP)技术,研究了稻麦轮作下农家肥(M)、氮肥+农家肥(NM)、氮磷肥+农家肥(NPM)、氮磷钾肥+农家肥(NPKM)、无肥(CK)、氮肥(N)、氮磷肥(NP)、氮磷钾肥(NPK)等不同施肥制度对石灰性紫色水稻土古菌群落结构的影响。研究结果表明,长期定位施肥明显影响土壤中的古菌组成。在长期施用氮肥+农家肥、氮磷肥和氮磷钾肥+农家肥处理的土壤中,古菌多样性指数低于农家肥、氮磷肥+农家肥、无肥、氮肥和氮磷钾肥处理。在DGGE图谱的基础上,分别选择种植水稻和小麦的氮磷钾肥处理土壤样品,对古菌克隆子的16SrDNA序列进行了系统发育分析,发现水稻土古菌与各种土壤及水体环境的古菌极其相似。对DGGE图谱的聚类分析发现,不管是种植水稻还是小麦,8种施肥处理的古菌都聚在3个群里。种植水稻时,M和NPK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(NPKM,NM,CK,N和NPM)聚成第三个群。种植小麦时,NPKM和M处理下的土壤古菌聚成一个群,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。聚类分析结果显示,作物类型会影响土壤古菌群落结构。
The quantity of soil microbes and the structure of ammonium oxidizing bacterial （AOB） community were analyzed using the dilution plate counting and most probable number method （MPN）, and denaturing gradient gel electrophoresis （DGGE）, respectively. Fertilizer application tended to increase the number of soil microbes and alter the AOB community compared to the control with no fertilizer application （CK）. Among the eight fertilizer treatments, soil samples from the treatments of mineral fertilizers （e.g., N, P, K） in combination with farmyard manure （M） had greater number.s of soil microbes and more complex structure of AOB community than those receiving mineral fertilizers alone. The principal component analyses （PCA） for ammonium oxidizing bacterial community structure showed that the eight fertilizer treatments could be divided into two PCA groups （PCA1 and PCA2）. For the soil sampled after rice harvest, PCA1 included NP, NM, NPM and NPKM fertilizer treatments, while PCA2 was consisted of CK, N, M and NPK fertilizer treatments. For soil samples collected after wheat harvest, PCA1 was consisted of M, NM, NPM and NPKM fertilizer treatments, while PCA2 was composed of CK, N, NP and NPK fertilizer treatments. For a given rotation, the richness of AOB community in PCA1 was greater than that in PCA2. In addition, AOB community structure was more complex in the soil after rice harvest than that after wheat harvest. The results indicated that different fertilizer treatments resulted in substantial changes of soil microbe number and AOB community. Furthermore, mineral fertilizers （N, NP, NPK） combined with farmyard manure were effective for increasing the quantity of soil microbes, enriching AOB community, and improving the soil biofertility.
