施肥旱作农田土壤中人类病原菌群落组成及动态变化特征

doi:10.3785/j.issn.1008-9209.2023.03.012

浙江大学学报（农业与生命科学版）

2024, Vol. 50

Issue (1): 98-108 DOI: 10.3785/j.issn.1008-9209.2023.03.012

资源利用与环境保护

施肥旱作农田土壤中人类病原菌群落组成及动态变化特征

齐明慧¹(

),程建华^1,²(

),唐翔宇^1,²

^1.浙江农林大学林业与生物技术学院，省部共建亚热带森林培育国家重点实验室，浙江杭州 311300
^2.中国科学院、水利部成都山地灾害与环境研究所，山地表生过程与生态调控重点实验室，四川成都 610041

Composition and dynamic change characteristics of human pathogenic communities in dryland farmland with manure application

Minghui QI¹(

),Jianhua CHENG^1,²(

),Xiangyu TANG^1,²

^1.State Key Laboratory of Subtropical Silviculture, College of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
^2.Key Laboratory of Mountain Surface Processes and Ecological Regulation, Institute of Mountain Hazards and Environment, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China

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摘要：

采集浙江省杭州、嘉兴、衢州、金华、龙游5个地区长期施用鸡粪、猪粪或化肥的农田土壤进行室内培养实验，然后采用高通量测序及序列比对方法分析供试土壤细菌和人类病原菌（human pathogenic bacteria, HPB）群落组成，旨在进一步探究施用粪肥后土壤中HPB群落组成差异及动态变化特征。结果表明：在160个供试土壤样品和2种粪肥样品中共比对到75种HPB，其中优势HPB为巨大芽孢杆菌（Bacillus_megaterium_QM_B1551，24.2%）和拜氏梭菌（Clostridium_beijerinckii_NCIMB_8052，23.1%）；施用猪粪处理的土壤中细菌和HPB的香农（Shannon）指数均有所降低，而施用鸡粪或化肥处理的土壤中细菌和HPB多样性无明显变化。主坐标分析结果表明，施用粪肥处理的土壤细菌群落组成与未施肥对照之间具有显著差异，尤其是施用猪粪处理差异最显著（P＜0.001）；土壤样品间共有HPB占所有HPB的22.7%；施用粪肥处理的土壤中大部分HPB的相对丰度高于未施肥对照，并随培养时间的延长而不断下降。方差分解分析结果表明，土壤理化性质、细菌群落及两者交互作用是土壤中HPB变异的重要因素。综上所述，施用粪肥处理的土壤中HPB的变化特征主要受肥料类型、土壤类型、土壤理化性质及固有细菌群落的影响。

关键词： 人类病原菌; 粪肥; 细菌群落; 土壤; 动态变化

Abstract:

To understand the community structure composition and dynamic change characteristics of human pathogenic bacteria (HPB) in soils after manure application, laboratory cultivation experiments were conducted on agricultural soils with long-term application of chicken manure, pig manure, or chemical fertilizer in five regions of Hangzhou, Jiaxing, Quzhou, Jinhua, and Longyou in Zhejiang Province, and the community compositions of the soil bacteria and HPB were analyzed by high-throughput sequencing and sequence alignment methods. The results showed that a total of 75 HPB were detected in 160 soil samples and two manure samples, and the dominant HPB were Bacillus_megaterium_QM_B1551 (24.2%) and Clostridium_beijerinckii_NCIMB_8052 (23.1%). The Shannon indexes of bacteria and HPB in the soils decreased after the application of pig manure, while the diversities of bacteria and HPB in the soils with the application of chicken manure or chemical fertilizer had no significant changes. The results of the principal coordinate analysis showed that there was a significant difference in the bacterial community composition of soils between the manure treatment and the unfertilized control, especially in the pig manure treatment (P＜0.001); 22.7% of all HPB were shared among the soil samples; and the relative abundance of most HPB in the soils treated with manure was higher than that in the unfertilized control, and it decreased continuously with the extension of cultivation time. The results of the variance partitioning analysis showed that soil physicochemical properties, bacterial communities, and their interactions were important factors contributing to the variation of HPB in the soils. In summary, the HPB variation characteristics in soils treated with manure are influenced mainly by manure types, soil types, soil physicochemical properties, and inherent bacterial communities.

Key words: human pathogenic bacteria manure bacterial community soil dynamic change

收稿日期: 2023-03-01 出版日期: 2024-03-01

CLC:

S154.3

基金资助: 国家自然科学基金项目(42007361);浙江农林大学科研发展基金项目(2020FR040);浙江省自然科学基金项目(LD21D010001)

通讯作者: 程建华 E-mail: 2020102022012@stu.zafu.edu.cn;chengjh@zafu.edu.cn

作者简介: 齐明慧（https://orcid.org/0009-0004-9190-284X），E-mail：2020102022012@stu.zafu.edu.cn

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引用本文:

齐明慧,程建华,唐翔宇. 施肥旱作农田土壤中人类病原菌群落组成及动态变化特征[J]. 浙江大学学报（农业与生命科学版）, 2024, 50(1): 98-108.

Minghui QI,Jianhua CHENG,Xiangyu TANG. Composition and dynamic change characteristics of human pathogenic communities in dryland farmland with manure application. Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 98-108.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2023.03.012 或 https://www.zjujournals.com/agr/CN/Y2024/V50/I1/98

原始土壤样品 Original soil sample		pH值 pH value	电导率 EC/ (μS/cm)	铵态氮 $N H 4 ＋$ -N/ (mg/kg)	硝态氮 $N O 3 －$ -N/ (mg/kg)	有效磷 AP/ (mg/kg)	总有机碳 TOC/ (mg/g)	全磷 TP/ (mg/g)	全氮 TN/ (mg/g)	全钾 TK/ (mg/g)
嘉兴市 JX	CM_Soil	7.54	103.55	12.27	24.91	71.29	20.76	2.50	1.68	6.89
	PM_Soil	8.08	132.25	19.80	42.11	129.56	26.64	1.34	1.81	6.92
	NPK_Soil	6.86	94.30	16.73	40.97	99.16	23.01	1.30	1.68	6.60
汤溪镇（金华市） TX	CM_Soil	5.57	63.75	15.05	28.98	750.88	22.23	0.99	1.32	4.96
	PM_Soil	6.45	120.90	19.10	62.23	1 457.04	24.08	0.60	1.42	5.25
	NPK_Soil	4.58	56.55	6.82	27.09	38.33	13.40	0.49	1.29	4.49
塘坞（衢州市） TW	CM_Soil	5.83	82.35	16.73	46.78	45.94	18.81	2.77	1.15	6.26
	PM_Soil	5.16	103.35	22.44	71.23	305.44	19.88	2.28	1.18	6.51
	NPK_Soil	7.96	142.50	4.99	90.94	30.09	23.32	4.44	1.11	5.72
昌化镇（杭州市） CH	CM_Soil	7.13	704.50	12.45	520.70	540.99	52.08	3.80	8.51	10.27
	PM_Soil	7.93	190.00	5.81	108.51	44.06	16.74	0.12	1.07	6.32
	NPK_Soil	5.33	66.95	9.01	25.82	201.19	15.01	0.11	0.98	6.84
龙游市 LY	CM_Soil	5.60	283.00	37.59	119.82	134.44	28.35	0.11	1.19	6.16
	PM_Soil	6.79	166.35	25.05	68.62	587.55	29.38	0.10	1.35	5.97
	1_NPK_Soil	4.89	38.95	14.73	13.65	136.32	6.65	0.55	0.57	7.46
	2_NPK_Soil	4.79	63.20	10.85	23.34	112.79	15.59	0.48	0.64	7.25

表1 原始土壤样品的基本理化性质

采样地 Sampling site	处理 Treatment	pH值pH value	电导率 EC/ (μS/cm)	铵态氮 $N H 4 ＋$ -N/ (mg/kg)	硝态氮 $N O 3 －$ -N/ (mg/kg)	总有机碳 TOC/ (mg/g)	有效磷 AP/ (mg/kg)	全磷 TP/ (mg/g)	全氮 TN/ (mg/g)
JX	CMs	6.89bc	543.60a	21.63a	292.04ab	32.00b	219.94b	1.79a	1.32a
	CMs_CK	7.22ab	194.70b	4.60b	121.82b	27.25c	101.28c	1.58b	1.08a
	PMs	7.37a	603.80a	11.69ab	415.64a	35.39a	369.72a	1.56b	1.59a
	PMs_CK	7.16ab	267.00b	3.86b	190.39b	31.12b	163.64bc	1.38c	1.44a
	NPKs	6.69c	224.20b	10.24ab	164.40b	26.71c	137.86c	1.55b	1.20a
	NPKs_CK	6.83bc	156.32b	3.56b	114.81b	27.07c	133.73c	1.59b	1.08a
TX	CMs	5.78c	386.40b	65.02a	122.35b	27.42b	886.57c	2.74c	1.09bc
	CMs_CK	4.97d	153.96d	14.97bc	123.80b	24.30c	756.81c	2.34d	0.82cd
	PMs	6.54a	531.40a	26.11b	342.07a	32.57a	1 251.15b	5.88a	2.03a
	PMs_CK	6.21b	251.18c	4.11c	191.70b	28.92b	1 529.93a	5.40b	1.54b
	NPKs	4.30e	146.48d	27.34b	80.17b	12.71d	29.95d	0.77e	0.41d
	NPKs_CK	4.46e	77.50e	2.54c	46.27b	13.23d	23.47d	0.80e	0.49d
TW	CMs	6.38b	460.60a	70.32a	146.86a	24.92b	223.50c	1.76c	1.45a
	CMs_CK	5.60c	139.84d	8.57c	122.18a	19.22c	72.01d	1.32e	0.89b
	PMs	6.00bc	372.80ab	36.93b	219.59a	26.63b	589.09a	2.40a	0.70b
	PMs_CK	5.00d	171.34d	15.13c	150.84a	20.41c	303.85b	1.98b	0.65b
	NPKs	7.30a	285.62bc	8.41c	201.68a	29.52a	60.72d	1.46d	0.61b
	NPKs_CK	7.46a	209.54cd	3.75c	146.36a	28.98a	63.51d	1.45d	0.62b
CH	CMs	7.05b	1 357.60a	686.60b	807.62a	223.17a	748.78a	5.41a	10.16a
	CMs_CK	7.05b	1 087.00a	1 087.00a	816.84a	213.78a	630.11b	4.82b	8.95b
	PMs	7.36ab	536.80b	465.09bc	315.12b	28.66b	223.51c	1.35c	0.86c
	PMs_CK	7.59a	248.60bc	248.60c	180.99b	25.78b	56.53d	1.03c	0.75c
	NPKs	4.94c	203.00bc	203.00c	127.35b	17.48b	202.02c	1.37c	0.69c
	NPKs_CK	5.10c	126.46c	126.46c	85.41b	15.75b	192.44c	1.35c	0.58c
LY	CMs	5.78b	846.20a	66.41a	363.84a	34.76b	307.26b	2.30b	1.25b
	CMs_CK	4.74c	389.80b	5.10c	295.64ab	25.55c	83.95b	1.55c	1.03bc
	PMs	6.82a	493.40b	25.74bc	302.12ab	38.86a	1 611.15a	6.79a	1.99a
	PMs_CK	6.55a	272.26c	14.78bc	183.70bc	33.65b	1 362.30a	6.73a	1.86a
	1_NPKs	4.68c	137.56d	39.32ab	92.86c	10.00f	135.69b	0.87d	0.36d
	1_NPKs_CK	4.85c	91.20d	13.59bc	61.58c	9.99f	125.05b	0.82d	0.33d
	2_NPKs	4.58c	163.08cd	29.10bc	89.95c	20.72d	98.09b	0.94d	0.39d
	2_NPKs_CK	4.68c	104.88d	18.54bc	65.86c	17.24e	88.90b	0.99d	0.54cd

表2 培养土壤样品的基本理化性质

图2 不同样品间属水平下共有和独有的HPB数量（A）及门水平下HPB组成（B）点线图表示集合间不同交集，单个节点表示特有的HPB，连线节点表示共有的HPB。

图3 不同采样地不同施肥处理下土壤细菌群落组成（A）和HPB组成（B）的差异

图4 施用鸡粪（A）、猪粪（B）和化肥（C）土壤中HPB的相对丰度

图5 土壤理化性质与细菌群落对HPB组成变化的影响

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