长期秸秆还田下土壤反硝化细菌群落的有机碳驱动机制

doi:10.3969/j.issn.1004-1524.2021.09.13

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (9): 1686-1699.DOI: 10.3969/j.issn.1004-1524.2021.09.13

长期秸秆还田下土壤反硝化细菌群落的有机碳驱动机制

贾生强¹^,²(), 范惠珊¹^,², 陈喜靖², 喻曼², 沈阿林², 苏瑶²^,^*()

1.浙江农林大学环境与资源学院,浙江杭州 311300
2.浙江省农业科学院环境资源与土壤肥料研究所,浙江杭州 310021

收稿日期:2020-12-20 出版日期:2021-09-25 发布日期:2021-10-09
通讯作者: 苏瑶
作者简介:* 苏瑶,E-mail: stellasu@sina.com
贾生强(1994—),男,山西大同人,硕士研究生,主要从事农业废弃物利用研究。E-mail: 164220950@qq.com
基金资助:
国家自然科学基金青年科学基金(41807034);浙江省自然科学基金(LQ18D030001);平湖市农业科技项目(PH2020004);国家小麦产业技术体系(CARS-3)

Driving mechanism of soil denitrifying bacterial community by soil organic carbon after long-term of straw return

JIA Shengqiang¹^,²(), FAN Huishan¹^,², CHEN Xijing², YU Man², SHEN Alin², SU Yao²^,^*()

1. College of Environment and Resources, Zhejiang A&F University, Hangzhou 311300, China
2. Institute of Environment, Resource, Soil and Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2020-12-20 Online:2021-09-25 Published:2021-10-09
Contact: SU Yao

摘要/Abstract

摘要：

采集连续5 a秸秆还田(SF)和不还田(CK)处理0~20、20~40、40~60、60~80、80~100 cm土层的土壤样品,对各层土壤不同有机碳、氮组分含量,以及反硝化细菌的丰度和种群组成进行分析。结果显示,SF处理0~40 cm土层的颗粒有机碳(POC)、20~60 cm土层的矿物结合态有机碳(MOC)和0~80 cm土层的全氮含量较CK处理分别显著(P<0.05)增加45.69%~142.75%、89.34%~272.68%和14.26%~90.34%,但0~40 cm土层的溶解性有机碳(DOC)和0~60 cm土层的微生物生物量碳(MBC)、硝态氮含量分别显著(P<0.05)减少68.89%~75.93%、35.58%~75.43%和12.91%~61.86%,其中,约63.81%的硝态氮损失发生在0~40 cm土层。相关性分析结果显示,土壤有机碳组分中的POC和MOC与土壤反硝化细菌的丰度显著(P<0.05)正相关,且影响其种群结构变化。SF处理0~60 cm土层nirS、nirK和nosZ基因拷贝数较CK处理增加2.5~6.7倍,并可有效促进unclassified_c_Betaproteobacteria(β-变形菌纲)、unclassified_f_Rhodocyclaceae(红环菌科)、unclassified_k_norank_d_Bacteria和unclassified_o_Burkholderiales(伯克氏菌目,属β-变形菌)的生长。综上,长期秸秆还田下,土壤反硝化细菌的生长及其种群结构变化主要受相对稳定的POC和MOC的驱动,引起的土壤硝态氮损失应在耕地肥力维系和提升,以及作物营养管理中予以必要考虑。

关键词: 秸秆还田, 溶解性有机碳, 颗粒有机碳, 矿物结合态有机碳, 反硝化细菌

Abstract:

In this study, soil samples in 0-20, 20-40, 40-60, 60-80 and 80-100 cm soil layer treated with 5 years of straw return (SF) and no straw return (CK) were collected, and the contents of different components of soil organic carbon and nitrogen, and the abundance and structure of soil denitrifying bacterial community were analyzed. The results showed that the contents of soil particulate organic carbon (POC) in 0-40 cm soil layer, mineral-associated organic carbon (MOC) in 20-60 cm soil layer and total nitrogen in 0-80 cm soil layer under SF treatment were significantly (P<0.05) increased by 45.69%-142.75%, 89.34%-272.68% and 14.26%-90.34%, respectively, compared with CK treatment, but the contents of soil dissolved organic carbon (DOC) in 0-40 cm soil layer, microbial biomass organic carbon (MBC) in 0-60 cm soil layer and nitrate nitrogen in 0-60 cm soil layer were significantly (P<0.05) decreased by 68.89%-75.93%, 35.58%-75.43% and 12.91%-61.86%, respectively. About 63.81% of nitrate nitrogen loss occurred in the 0-40 cm soil. Correlation analysis results showed that soil POC and MOC were significantly (P<0.05) positively correlated with the abundance of soil denitrifying bacterial community, and affected the community structure. The number of nirS、nirK and nosZ gene copies in the 0-60 cm soil layer under SF treatment was 2.5-6.7 times higher than that under CK, and the genera of unclassified_c_Betaproteobacteria, unclassified_f_Rhodocyclaceae, unclassified_k_norank_d_Bacteria and unclassified_o_Burkholderiales were enhanced by the long-term straw return. In general, the abundance and structure of soil denitrifying bacterial community under long-term straw return were mainly driven by the relatively stable soil organic carbon components, such as POC and MOC. The loss of soil nitrate nitrogen caused by the enhanced soil denitrification should be taken into consideration in the maintenance and promotion of cultivated land fertility and crop nutrition management in future.

Key words: straw return, dissolved organic carbon, particulate organic carbon, mineral-associated organic carbon, denitrifying bacterial community

中图分类号:

S154

贾生强, 范惠珊, 陈喜靖, 喻曼, 沈阿林, 苏瑶. 长期秸秆还田下土壤反硝化细菌群落的有机碳驱动机制[J]. 浙江农业学报, 2021, 33(9): 1686-1699.

JIA Shengqiang, FAN Huishan, CHEN Xijing, YU Man, SHEN Alin, SU Yao. Driving mechanism of soil denitrifying bacterial community by soil organic carbon after long-term of straw return[J]. Acta Agriculturae Zhejiangensis, 2021, 33(9): 1686-1699.

图/表 11

表1 荧光实时定量PCR扩增引物

Table 1 Primers used in fluorescent real-time quantitative PCR amplification

目的基因 Target gene	引物 Primer	引物序列 Primer sequence	参考文献 Reference
nirK	F1aCu	ATC ATG GTSCTG CCG CG	Krishnani^[29]
	R3Cu	GCC TCG ATC AGR TTG TGG TT
nirS	cd3aF	GTS AAC GTS AAG GAR ACS GG	Szukics等^[30]
	R3cdR	GAS TTC GGR TGS GTC TTG A
nosZ	NosF2	GGG CTB GGG CCR TTG CA	Li等^[31]
	NosR2	GAA GCG RTC CTT SGA RAA CTT G

图1 秸秆还田(SF)和不还田(CK)处理不同土层的DOC、MBC、POC和MOC含量柱右侧无相同小写字母的表示差异显著(P<0.05)。DOC,溶解性有机碳;MBC,微生物生物量碳;POC,颗粒有机碳;MOC,矿物结合态有机碳。下同。

Fig.1 DOC, MBC, POC and MOC contents in different soil layers under straw return (SF) and control (CK) Bars marked without the same letters indicated significant difference at P<0.05. DOC, Dissolved organic carbon; MBC, Microbial biomass organic carbon; POC, Particulate organic carbon; MOC, Mineral-assoiciated organic carbon. The same as below.

图2 秸秆还田(SF)和不还田(CK)处理不同土层的全氮、碱解氮、铵态氮和硝态氮含量 TN,全氮;AN,碱解氮; NH 4 +-N,铵态氮; NO 3 --N,硝态氮。下同。

Fig.2 Contents of total nitrogen, alkali-hydrolyzable nitrogen, ammoniacal nitrogen and nitrate nitrogen in different soil layers under straw return (SF) and control (CK) TN, Total nitrogen; AN, Alkali-hydrolyzable nitrogen; NH 4 +-N, Ammoniacal nitrogen; NO 3 --N, Nitrate nitrogen. The same as below.

图3 秸秆还田处理土壤全氮与土壤MOC、POC含量的关系

Fig.3 Relationship between soil total nitrogen and soil MOC or POC under straw return

图4 秸秆还田(SF)和不还田(CK)处理不同土层的nirS、nirK和nosZ基因拷贝数

Fig.4 Copy number of nirS, nirK and nosZ gene in different soil layers under straw return (SF) and control (CK)

表2 不同反硝化细菌功能基因拷贝数与土壤有机质、氮组分的相关性

Table 2 Correlation within gene copy number of denitrifying bacteria and contents of soil organic carbon and nitrogen components

基因 Gene	土层深度 Soil depth/cm	DOC	MBC	MOC	POC	$NH 4 +$ -N	$NO 3 -$ -N	AN	TN
nirS	0~20	-0.979^**	-0.980^**	0.386	0.915^*	-0.978^**	-0.879^*	-0.835^*	0.942^**
	20~40	-0.968^**	-0.974^**	0.971^**	0.982^**	-0.623	-0.979^**	-0.960^**	0.991^**
	40~60	-0.940^**	-0.992^**	0.992^**	0.994^**	0.973^**	-0.988^**	-0.970^**	0.979^**
nirK	0~20	-0.952^**	-0.938^**	0.523	0.830^*	-0.947^**	-0.851^*	-0.722	0.901^*
	20~40	-0.977^**	-0.984^**	0.987^**	0.959^**	-0.528	-0.975^**	-0.977^**	0.965^**
	40~60	-0.966^**	-0.972^**	0.980^**	0.944^**	0.956^**	-0.981^**	-0.977^**	0.989^**
nosZ	0~20	-0.982^**	-0.984^**	0.347	0.918	-0.991^**	-0.838^*	-0.827^*	0.896^*
	20~40	-0.974^**	-0.985^**	0.985^**	0.981^**	-0.567	-0.985^**	-0.969^**	0.995^**
	40~60	-0.941^**	-0.979^**	0.985^**	0.955^**	0.953^**	-0.979	-0.969^**	0.985^**

表2 不同反硝化细菌功能基因拷贝数与土壤有机质、氮组分的相关性

Table 2 Correlation within gene copy number of denitrifying bacteria and contents of soil organic carbon and nitrogen components

基因 Gene	土层深度 Soil depth/cm	DOC	MBC	MOC	POC	$NH 4 +$ -N	$NO 3 -$ -N	AN	TN
nirS	0~20	-0.979^**	-0.980^**	0.386	0.915^*	-0.978^**	-0.879^*	-0.835^*	0.942^**
	20~40	-0.968^**	-0.974^**	0.971^**	0.982^**	-0.623	-0.979^**	-0.960^**	0.991^**
	40~60	-0.940^**	-0.992^**	0.992^**	0.994^**	0.973^**	-0.988^**	-0.970^**	0.979^**
nirK	0~20	-0.952^**	-0.938^**	0.523	0.830^*	-0.947^**	-0.851^*	-0.722	0.901^*
	20~40	-0.977^**	-0.984^**	0.987^**	0.959^**	-0.528	-0.975^**	-0.977^**	0.965^**
	40~60	-0.966^**	-0.972^**	0.980^**	0.944^**	0.956^**	-0.981^**	-0.977^**	0.989^**
nosZ	0~20	-0.982^**	-0.984^**	0.347	0.918	-0.991^**	-0.838^*	-0.827^*	0.896^*
	20~40	-0.974^**	-0.985^**	0.985^**	0.981^**	-0.567	-0.985^**	-0.969^**	0.995^**
	40~60	-0.941^**	-0.979^**	0.985^**	0.955^**	0.953^**	-0.979	-0.969^**	0.985^**

表3 nirS型反硝化细菌多样性指数

Table 3 Diversity index of nirS-type denitrifying bacterial community

处理 Treatment	土层深度 Soil depth/cm	覆盖度 Coverage/%	多样性Diversity
处理 Treatment	土层深度 Soil depth/cm	覆盖度 Coverage/%	Shannon	Simpson	ACE	Chao
CK	0~20	98.49	3.84±0.02 ab	0.066±0.005 a	563±7 ab	560±8 c
	20~40	98.47	3.80±0.09 b	0.068±0.002 a	553±10 b	549±12 c
	40~60	99.29	3.95±0.14 ab	0.038±0.001 c	455±23 c	456±13 d
SF	0~20	98.29	4.09±0.01 ab	0.045±0.002 bc	635±23 a	616±2 b
	20~40	98.38	4.09±0.06 ab	0.060±0.001 ab	640±16 a	673±14 a
	40~60	98.67	4.19±0.04 a	0.045±0.004 bc	543±16 b	544±7 c

图5 不同处理0~60 cm土层nirS型反硝化细菌群落的主坐标分析结果空心图标代表CK,实心图标代表SF,D1、D2、D3分别代表0~20、20~40、40~60 cm土层。下同。PC1,第1主成分;PC2,第2主成分。

Fig.5 Principal coordinate analysis of nirS-type denitrifying bacterial community in 0-60 cm soil layer under different treatments Hollow icons represented CK; solid icons represented SF. D1, D2 and D3 represented soil layers of 0-20, 20-40, 40-60 cm, respectively. The same as below. PC1, The 1st principle component; PC2, The 2nd principle component.

图6 不同处理0~60 cm土层nirS型反硝化细菌种群在属水平上的组成

Fig.6 Community composition of nirS-type denitrifying bacterial at genus level in 0-60 cm soil layer under different treatments

图7 0~60 cm土层不同处理间存在显著(P<0.05)差异的nirS型反硝化菌属

Fig.7 nirS-type denitrifying bacteria with significant (P<0.05) difference under different treatments in 0-60 cm soil layer

图8 影响nirS型反硝化细菌种群结构的冗余分析结果 RD1,主成分1;RD2,主成分2。

Fig.8 Redundancy analysis result of nirS-type denitrifying bacterial community structure RD1, The 1st principle component; RD2, The 2nd principle component.

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长期秸秆还田下土壤反硝化细菌群落的有机碳驱动机制

Driving mechanism of soil denitrifying bacterial community by soil organic carbon after long-term of straw return

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