长期定位施肥对石灰性紫色水稻土 古菌群落结构的影响

doi:10.3724/SP.J.1003.2011.08193

生物多样性 ›› 2011, Vol. 19 ›› Issue (3): 369-376. DOI: 10.3724/SP.J.1003.2011.08193

所属专题：土壤生物与土壤健康

长期定位施肥对石灰性紫色水稻土古菌群落结构的影响

辜运富¹, 张小平¹^,^*(), 涂仕华², Kristina Lindström³

1 (四川农业大学资源环境学院微生物系, 四川雅安 625014)
2 (四川省农业科学院土壤肥料研究所, 成都 610066)

收稿日期:2010-08-06 接受日期:2011-02-05 出版日期:2011-05-20 发布日期:2013-12-10
通讯作者: 张小平
作者简介:*E-mail:aumdwsb@sicau.edu.cn
基金资助:
国家科技支撑计划(2006BAD05B06);国家科技支撑计划(2006BAD 02A 14-13)

Effect of long-term fertilization on archaeal community structure in calcareous purplish paddy soil

Yunfu Gu¹, Xiaoping Zhang¹^,^*(), Shihua Tu², Kristina Lindström³

1 College of Resource and Environment, Sichuan Agricultural University, Ya’an 625014, China
2 Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences, Chengdu 610066, China

Received:2010-08-06 Accepted:2011-02-05 Online:2011-05-20 Published:2013-12-10
Contact: Xiaoping Zhang

摘要/Abstract

摘要：

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16S rDNA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。

关键词: 施肥制度, DGGE, RFLP, 古菌群落

Abstract

To better understand the role that fertilization systems play in soil fertility buildup and evolution, we employed denaturing gradient gel electrophoresis (DGGE) and restriction fragment length polymorphism (RFLP) to determine the influence of different fertilization treatments on archaeal community structure in calcareous purplish paddy soil under rice/wheat rotations. The eight fertilization treatments were as follows: manure only (M), nitrogen plus manure (NM), nitrogen and phosphorus plus manure (NPM), nitrogen, phosphorus and potassium plus manure (NPKM), without fertilization (CK), mineral nitrogen (N), nitrogen-phosphorus (NP) and nitrogen, and phosphorus and potassium (NPK). Our results showed that long-term fertilization significantly affected soil archaeal community structure; the richness and diversity of archaeal community under NM, NP and NPKM were lower than those under the other fertilization treatments (M, NPM, CK, N, and NPK). Based on the DGGE patterns, two soil DNA samples isolated from the NPK-amended soil were used for RFLP analysis of archaea. Phylogenetic analyses showed that archaea in the calcareous purplish paddy soil was highly diverse, and the sequences were closely related to those archaeal sequences isolated from various soils and water environment. Cluster analysis of the DGGE profiles showed that archaeal communities under the eight fertilization treatments clustered into three groups. In soil from paddies currently under rice cultivation, the archaeal communities in the soil amended with M and NPK grouped into the first cluster, while NP was in the second group, and NPKM, NM, CK, N and NPM were in the third. In the soil with wheat cultivation, NP-treated archaeal communities clustered into a cluster, NPKM and M were in the second cluster, and N, NPK, NM, NPM and CK soil communities comprised the third cluster. The cluster analysis showed that crop type impacts the community structure of soil archaea.

Key words: fertilization system, DGGE, RFLP, archaeal communities

辜运富, 张小平, 涂仕华, Kristina Lindström (2011) 长期定位施肥对石灰性紫色水稻土古菌群落结构的影响. 生物多样性, 19, 369-376. DOI: 10.3724/SP.J.1003.2011.08193.

Yunfu Gu, Xiaoping Zhang, Shihua Tu, Kristina Lindström (2011) Effect of long-term fertilization on archaeal community structure in calcareous purplish paddy soil. Biodiversity Science, 19, 369-376. DOI: 10.3724/SP.J.1003.2011.08193.

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https://www.biodiversity-science.net/CN/Y2011/V19/I3/369

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参考文献 27

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	施肥方式 Fertilization treatments
	M	NM	NPM	NPKM	CK	N	NP	NPK
水稻收获后 After rice cultivation	4.043	3.993	4.987	4.057	3.969	4.990	3.942	3.753
小麦收获后 After wheat cultivation	4.855	4.887	4.232	4.209	4.173	4.783	4.075	4.787

	施肥方式 Fertilization treatments
	M	NM	NPM	NPKM	CK	N	NP	NPK
水稻收获后 After rice cultivation	4.043	3.993	4.987	4.057	3.969	4.990	3.942	3.753
小麦收获后 After wheat cultivation	4.855	4.887	4.232	4.209	4.173	4.783	4.075	4.787

引物 Primer	16S rDNA目标区(碱基数) 16S rDNA target (base number)	引物序列 Primer sequence
PRA46F	Archaea (46-60)	5′- C/TTAAGCCATGCG/AAGT-3′
PREA1100R	Archaea (1100-1117)	5′- T/CGGGTCTCGCTCGTTG/ACC-3′
PARCH340F-GC	Archaea V3 region (340-358)	5′- CCTACGGGGC/TGCAG/CCAG-3′
PARCH519R	Archaea V3 region (519-534)	5′- TTACCGCGGCG/TGCTG-3′

引物 Primer	16S rDNA目标区(碱基数) 16S rDNA target (base number)	引物序列 Primer sequence
PRA46F	Archaea (46-60)	5′- C/TTAAGCCATGCG/AAGT-3′
PREA1100R	Archaea (1100-1117)	5′- T/CGGGTCTCGCTCGTTG/ACC-3′
PARCH340F-GC	Archaea V3 region (340-358)	5′- CCTACGGGGC/TGCAG/CCAG-3′
PARCH519R	Archaea V3 region (519-534)	5′- TTACCGCGGCG/TGCTG-3′

引物 Primers	16S rDNA目标区(碱基数) 16S rDNA target (base number)	引物序列 Primer sequence	参考文献 References
Ar4F	Archaea (8-25)	5′- TCY GGT TGA TCC TGC CRG-3′	Hershberger et al., 1996
Ar958R	Archaea (958-967)	5′- YCC GGC GTT GAV TCC AAT T-3′	Delong, 1992
Ar9R	Archaea (906-927)	5′-CCC GCC AAT TCC TTT AAG TTT C-3′	Jurgens et al., 1997
Ar3F	Archaea (7-26)	5′- TTC CGG TTG ATC CTG CCG GA-3′	Jurgens et al., 1997

长期定位施肥对石灰性紫色水稻土古菌群落结构的影响

Effect of long-term fertilization on archaeal community structure in calcareous purplish paddy soil

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施肥处理 Fertilization treatments	Shannon多样性指数 Shannon's diversity index (H)		丰富度 Richness (S)		均匀度 Evenness (EH)
施肥处理 Fertilization treatments	水稻 Rice	小麦 Wheat	水稻 Rice	小麦 Wheat	水稻 Rice	小麦 Wheat
M	3.08±0.13^a	3.13±0.14^a	21±2.00^a	20±2.00^a	1.016±0.00^a	1.014±0.00^a
NM	2.94±0.09^a	2.99±0.07^ab	18±1.15^b	19±1.53^ab	1.017±0.00^a	1.018±0.01^a
NPM	3.04±0.09^a	3.09±0.09^a	20±1.53^a	21±1.53^a	1.014±0.05^a	1.015±0.00^a
NPKM	2.99±0.09^a	2.99±0.03^ab	19±0.58^b	19±1.53^ab	1.017±0.05^a	1.017±0.01^a
CK	3.09±0.07^a	3.04±0.07^ab	21±1.16^a	20±1.15^a	1.015±0.05^a	1.015±0.00^a
N	2.99±0.09^a	3.08±0.03^ab	19±0.06^b	21±1.53^a	1.014±0.00^a	1.017±0.00^a
NP	2.89±0.09^b	2.89±0.07^b	17±1.15^b	17±1.53^b	1.019±0.00^a	1.020±0.01^a
NPK	3.13±0.16^a	3.04±0.11^ab	22±2.08^a	20±3.00^a	1.013±0.03^a	1.014±0.00^a

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长期定位施肥对石灰性紫色水稻土 古菌群落结构的影响

Effect of long-term fertilization on archaeal community structure in calcareous purplish paddy soil

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长期定位施肥对石灰性紫色水稻土古菌群落结构的影响