硫红霉素菌渣有机肥对大豆土壤中耐药菌及相关抗性基因的影响

doi:10.6048/j.issn.1001-4330.2023.02.020

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (2): 424-431.DOI: 10.6048/j.issn.1001-4330.2023.02.020

• 园艺特产·植物保护·微生物·土壤肥料·节水灌溉 • 上一篇下一篇

硫红霉素菌渣有机肥对大豆土壤中耐药菌及相关抗性基因的影响

易鸳鸯¹^,²^,³(), 谢芳¹^,², 田世英¹^,², 张志东³(), 顾美英³, 彭小武¹^,²()

1.新疆环境保护科学研究院,乌鲁木齐 830011
2.新疆环境污染监控与风险预警重点实验室/新疆清洁生产工程技术研究中心/国家环境保护准噶尔荒漠绿洲交错区科学观测研究站,乌鲁木齐 830011
3.新疆农业科学院微生物应用研究所/新疆特殊环境微生物实验室,乌鲁木齐 830091

出版日期:2023-02-20 发布日期:2023-03-31
通信作者: 张志东(1977-),男,新疆乌鲁木齐人,研究员,研究方向为特殊环境微生物,(E-mail)zhangzheedong@qq.com;
彭小武(1969-),女,湖南新邵人,高级工程师,研究方向为环境污染物风险与控制,(E-mail)437848836@qq.com
作者简介:易鸳鸯(1990-),女,新疆乌鲁木齐人,工程师,研究方向为环境微生物,(E-mail)15022970169@163.com
基金资助:
自治区科技专项-公益性科研院所基本科研业务专项经费资助(KY2020080);新疆农业科学院农业科技创新平台能力提升建设专项(nkypt005);新疆农业科学院自主培育团队建设专项(nkyzztd-001)

Effect of Organic Fertilizer of Thioerythromycin Residue on Soybean Soil Drug-resistant Bacteria and Their Resistance Genes

YI Yuanyang¹^,²^,³(), XIE Fang¹^,², TIAN Shiying¹^,², ZHANG Zhidong³(), GU Meiying³, PENG Xiaowu¹^,²()

1. Xinjiang Academy of Environmental Protection Sciences, Urumqi 830011, China
2. Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
3. Xinjiang Key Laboratory for Environmental Pollution Monitoring and Risk Warning/Xinjiang Engineering Technology Research Center for Cleaner Production/Junggar Desert-oasis Ecotone Station for Scientific Observation and Research of National Environmental Protection, Urumqi 830011, China

Published:2023-02-20 Online:2023-03-31
Correspondence author: ZHANG Zhidong(1977-),male,from Urumqi,Xinjiang,researcher. research field: Environmental microorganism, (E-mail) zhangzheedong@qq.com;
PENG Xiaowu(1969-),female, from Xinshao, Hunan, senior engineer, research field:risk and control of environmental pollutants, (E-mail) 437848836@qq.com
Supported by:
Basic Scientific R &D Program of Public Welfare Research Institutions of Xinjiang Uygur Autonomous Region of China(KY2020080);Agricultural Science and Technology Innovation Platform Capacity Improvement Project of Xinjiang Academy of Agricultural Sciences(nkypt005);Team Building Special Fund for Independent Cultivation Program of Xinjiang Academy of Agricultural Sciences(nkyzztd-001)

摘要/Abstract

摘要：

【目的】 研究硫红霉素菌渣有机肥施用后对大豆农田土壤中耐药菌及相关抗性基因丰度的影响,为硫红霉素菌渣有机肥施用生物安全性评价提供科学依据。【方法】 检测不同菌渣有机肥施用量下,大豆不同生长时期土壤中耐药细菌的数量和种类,并采用荧光定量PCR方法检测相关抗性基因(ARGs)污染水平,分析菌渣有机肥施用对耐药菌及相关抗性基因的影响。【结果】 大豆苗期土壤中各类细菌数量大于结果期,苗期样品中土壤细菌总数、硫红霉素抗性菌数显著高于对照组,而青霉素、头孢拉定抗性菌株数与对照组差异不显著。在结果期样品中,施加了硫红霉素药渣有机肥的土壤中各类菌落总数与对照组差异不显著。所获得14株硫红霉素耐药菌菌株分布于11个菌属,其中假节杆菌属、芽胞杆菌属、类谷氨酸杆菌属菌株数占总菌株数的比例最高,25株青霉素耐药菌菌株分布于7个菌属,其中链霉菌属菌株数最高。12株头孢拉定耐药菌分布于5个菌属,其中假单胞菌属菌株数占比例达到50.00%。施用硫红霉素菌渣有机肥对土壤中常见ARGs的绝对丰度和相对丰度有一定的影响,但影响不显著。【结论】 硫红霉素菌渣有机肥施用后对作物土壤中ARGs水平没有显著影响。

关键词: 硫红霉素菌渣; 大豆土壤; 耐药菌; ARGs; q-PCR

Abstract:

【Objective】 To explore the effect of thioerythromycin residue organic fertilizer on the abundance of drug-resistant bacteria and related resistance genes in soybean farmland soil, and provide a scientific basis for the biosafety evaluation of the fertilizer. 【Methods】 By detecting the number and species of drug-resistant bacteria in soil in different growth stages of soybean under different application rates of bacterial residue organic fertilizer, and detecting the pollution level of related resistance genes (args) by fluorescence quantitative PCR, the effects of bacterial residue organic fertilizer on drug-resistant bacteria and related resistance genes were analyzed. 【Results】 The total number of soil bacteria and the number of thioerythromycin resistant bacteria in soybean seedling stage were significantly higher than those in the control group, while the number of penicillin and Cefradine resistant bacteria was not significantly different from the control group.In the result period, there was no significant difference between the total number of various colonies in the soil with thioerythromycin residue organic fertilizer and the control group.The obtained 14 strains of thioerythromycin resistant bacteria were distributed in 11 genera, of which pseudoarthrobacteria, bacillus and glutamate like bacteria accounted for the highest proportion in the total number of bacteria, and 25 strains of penicillin resistant bacteria were distributed in 7 genera, of which Streptomyces had the highest number.12 strains of cefradine resistant bacteria were distributed in 5 genera, of which Pseudomonas was the highest, accounting for 50.00% of the total.The application of thioerythromycin residue organic fertilizer had a certain effect on the absolute and relative abundance of common args in soil, but the effect was not significant. 【Conclusion】 The application of thioerythromycin residue organic fertilizer has no significant effect on the level of args in crop soil.

Key words: thioerythromycin residue; soybean soil; drug-resistant bacteria; ARGs; q-PCR

中图分类号:

S188

易鸳鸯, 谢芳, 田世英, 张志东, 顾美英, 彭小武. 硫红霉素菌渣有机肥对大豆土壤中耐药菌及相关抗性基因的影响[J]. 新疆农业科学, 2023, 60(2): 424-431.

YI Yuanyang, XIE Fang, TIAN Shiying, ZHANG Zhidong, GU Meiying, PENG Xiaowu. Effect of Organic Fertilizer of Thioerythromycin Residue on Soybean Soil Drug-resistant Bacteria and Their Resistance Genes[J]. Xinjiang Agricultural Sciences, 2023, 60(2): 424-431.

图/表 9

参考文献 24

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处理 Treat ments	红霉素残留 Erythromycin residue (mg/kg)	pH	有机质 OM (%)	全磷 Total P (mg/kg)	速效磷 Available P (mg/kg)	全钾 Total K (mg/kg)	速效钾 Available K (mg/kg)	全氮 Total N (mg/kg)
A₁	ND	8.37	14 400	221.0	4.89	4 110	235	140
A₂	ND	8.60	12 400	139.0	3.32	1 810	198	200
CK	ND	8.34	11 600	188.0	3.35	3 990	231	180

处理 Treat ments	红霉素残留 Erythromycin residue (mg/kg)	pH	有机质 OM (%)	全磷 Total P (mg/kg)	速效磷 Available P (mg/kg)	全钾 Total K (mg/kg)	速效钾 Available K (mg/kg)	全氮 Total N (mg/kg)
A₁	ND	8.37	14 400	221.0	4.89	4 110	235	140
A₂	ND	8.60	12 400	139.0	3.32	1 810	198	200
CK	ND	8.34	11 600	188.0	3.35	3 990	231	180

引物名称 Primer name	序列(5'-3') Suquence
ermB-F	GGATATTCACCGAACACTAGGG
ermB-R	TGGAACATCTGTGGTATGGC
ermF-F	CGACACAGCTTTGGTTGAAC
ermF-R	GGACCTACCTCATAGACAAG
blaTEM-F	TCGCCGCATACACTATTCTCAGAATGA
blaTEM-R	ACGCTCACCGGCTCCAGATTTAT
sul1-F	GCCGATGAGATCAGACGTATTG
sul1-R	GAAGCTGTCGATTGAAACACG
sul2-F	AACCGCCTTGTCCTTGATC
sul2-R	CAGCCGCAATTCATCGAAC

引物名称 Primer name	序列(5'-3') Suquence
ermB-F	GGATATTCACCGAACACTAGGG
ermB-R	TGGAACATCTGTGGTATGGC
ermF-F	CGACACAGCTTTGGTTGAAC
ermF-R	GGACCTACCTCATAGACAAG
blaTEM-F	TCGCCGCATACACTATTCTCAGAATGA
blaTEM-R	ACGCTCACCGGCTCCAGATTTAT
sul1-F	GCCGATGAGATCAGACGTATTG
sul1-R	GAAGCTGTCGATTGAAACACG
sul2-F	AACCGCCTTGTCCTTGATC
sul2-R	CAGCCGCAATTCATCGAAC

生育期 Growth period	处理 Treat ments	总细菌数 Total bacterias (10⁷CFU/g)	硫红霉素耐药菌 Thioerythromycin resistant bacteria (10³CFU/g)	青霉素耐药菌 Penicillin resistant bacteria(10³CFU/g)	头孢拉定耐药菌 Cefradine resistant bacteria(10⁵CFU/g)
苗期 Seedling stage	A₁	2.43±0.35^a	4.90±0.42^a	5.24±0.54^a	2.55±0.75^a
	A₂	2.52±0.23^a	5.50±0.16^a	5.30±0.36^a	2.57±0.57^a
	CK	1.8 7±0.13^b	4.33±0.25^b	6.00±0.33^a	2.70±0.54^a
结果期 Result period	A₁	1.76±0.82^a	3.67±0.82^a	4.00±0.38^a	0.80±1.65^a
	A₂	1.53±0.43^a	4.53±0.14^a	4.30±0.25^a	1.02±0.45^a
	CK	1.82±0.23^a	3.51±0.81^a	40.6±0.11^a	1.00±0.64^a

硫红霉素菌渣有机肥对大豆土壤中耐药菌及相关抗性基因的影响

Effect of Organic Fertilizer of Thioerythromycin Residue on Soybean Soil Drug-resistant Bacteria and Their Resistance Genes

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耐药菌类型 Type of resistant bacteria	菌属 Genus	菌株数 Number of bacteria	所占比例 Propo rtion (%)
硫红霉素耐药菌 Thioery thromycin resistant bacteria	Paenibacillus lautus	1	7.14
	Lysinibacillus macroides	1	7.14
	Pseudarthrobacter siccitolerans	2	14.29
	Bacillus idriensis	2	14.29
	Alcaligenes faecalis	1	7.14
	Sinorhizobium meliloti	1	7.14
	Paeniglutamicibacter sulfureus	2	14.29
	Planomicrobium glaciei	1	7.14
	Microbacterium profundi	1	7.14
	Arthrobacter pascens	1	7.14
	Brevibacillus halotolerans	1	7.14
青霉素耐药菌 Penicillin resistant bacteria	Streptomyces ramulosus	18	72.00
	Amycolatopsis roodepoortensis	1	4.00
	Proteus	1	4.00
	Bacillus idriensis	2	8.00
	Rhodococcus jostii	1	4.00
	Chitinophaga lutea	1	4.00
	Parapedobacter pyrenivorans	1	4.00
头孢拉定耐药菌 Cefradine resistant bacteria	Streptomyces ramulosus	2	16.67
	Pseudomonas laurentiana	6	50.00
	Microbacterium shaanxiense	1	8.33
	Bacillus idriensis	2	16.67
	[Brevibacterium] frigoritolerans	1	8.33

序号 NO.	抗性基因 ARGs	线性方程 linear algebraic equations	相关系数 R² correlation coefficient
1	ermB	Y=-2.985 4x+35.026	0.990 5
2	ermF	Y=-2.757 1x+31.918	0.993 9
3	blaTEM	Y=-2.945 7x+31.698	0.994 2
4	sul1	Y=-3.006 1x+34.948	0.994 7
5	sul2	Y=-2.337 2x+35.547	0.991 8
6	16s rDNA V4	Y=-2.881 9x+36.494	0.992 1

处理 Treat ments	生育期 Growth period	ErmB (10³)	ErmF (10³)	BlaTEM (10⁴)	sul1 (10³)	sul2 (10³)	16S rDNA (10⁷)
出苗期 Seedling stage	A₁	4.06±1.85^a	4.08±2.11^a	4.13±1.71^a	3.12±3.01^a	3.89±2.27^a	2.79±1.27^a
	A₂	5.60±0.85^a	5.36±0.61^a	4.57±0.75^a	5.75±0.31^a	5.61±0.56^a	3.71±0.33^a
	CK	3.35±2.95^a	3.31±3.01^a	5.46±0.53^a	4.86±1.09^a	4.85±1.37^a	1.63±1.47^a
结果期 Result period	A₁	1.27±0.95^a	2.28±0.21^a	2.35±0.31^a	2.53±1.15^a	2.66±1.51^a	1.72±0.45^a
	A₂	1.34±0.75^a	2.18±0.42^a	2.52±0.23^a	3.65±0.17^a	3.81±0.43^a	1.64±0.51^a
	CK	1.29±1.03^a	2.01±0.57^a	2.41±0.32^a	2.92±0.87^a	2.71±1.45^a	1.84±0.44^a

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