Xinjiang Agricultural Sciences ›› 2023, Vol. 60 ›› Issue (2): 424-431.DOI: 10.6048/j.issn.1001-4330.2023.02.020
• Horticultural Special Local Products·Plant Protection·Microbes·Soil Fertilizer· Water Saving Irrigation • Previous Articles Next Articles
YI Yuanyang1,2,3(), XIE Fang1,2, TIAN Shiying1,2, ZHANG Zhidong3(), GU Meiying3, PENG Xiaowu1,2()
Online:
2023-02-20
Published:
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:
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()
通讯作者:
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:
CLC Number:
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.
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.
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URL: http://www.xjnykx.com/EN/10.6048/j.issn.1001-4330.2023.02.020
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 |
Table 1 Residue thioerythromycin and physical-chemical properties of different treatments soil
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 |
Table 2 Primer sequences of antibiotic resitance genes
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±0.35a | 4.90±0.42a | 5.24±0.54a | 2.55±0.75a |
A2 | 2.52±0.23a | 5.50±0.16a | 5.30±0.36a | 2.57±0.57a | |
CK | 1.8 7±0.13b | 4.33±0.25b | 6.00±0.33a | 2.70±0.54a | |
结果期 Result period | A1 | 1.76±0.82a | 3.67±0.82a | 4.00±0.38a | 0.80±1.65a |
A2 | 1.53±0.43a | 4.53±0.14a | 4.30±0.25a | 1.02±0.45a | |
CK | 1.82±0.23a | 3.51±0.81a | 40.6±0.11a | 1.00±0.64a |
Table 3 Drug resistant bacteria in soil at different treatments and growth stages of soybean
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±0.35a | 4.90±0.42a | 5.24±0.54a | 2.55±0.75a |
A2 | 2.52±0.23a | 5.50±0.16a | 5.30±0.36a | 2.57±0.57a | |
CK | 1.8 7±0.13b | 4.33±0.25b | 6.00±0.33a | 2.70±0.54a | |
结果期 Result period | A1 | 1.76±0.82a | 3.67±0.82a | 4.00±0.38a | 0.80±1.65a |
A2 | 1.53±0.43a | 4.53±0.14a | 4.30±0.25a | 1.02±0.45a | |
CK | 1.82±0.23a | 3.51±0.81a | 40.6±0.11a | 1.00±0.64a |
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 |
Table 4 Genus distribution of drug resistant bacteria isolated from soybean soil
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 |
Table 5 Standard curve equation of ARGs and 16S rDNA
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±1.85a | 4.08±2.11a | 4.13±1.71a | 3.12±3.01a | 3.89±2.27a | 2.79±1.27a |
A2 | 5.60±0.85a | 5.36±0.61a | 4.57±0.75a | 5.75±0.31a | 5.61±0.56a | 3.71±0.33a | |
CK | 3.35±2.95a | 3.31±3.01a | 5.46±0.53a | 4.86±1.09a | 4.85±1.37a | 1.63±1.47a | |
结果期 Result period | A1 | 1.27±0.95a | 2.28±0.21a | 2.35±0.31a | 2.53±1.15a | 2.66±1.51a | 1.72±0.45a |
A2 | 1.34±0.75a | 2.18±0.42a | 2.52±0.23a | 3.65±0.17a | 3.81±0.43a | 1.64±0.51a | |
CK | 1.29±1.03a | 2.01±0.57a | 2.41±0.32a | 2.92±0.87a | 2.71±1.45a | 1.84±0.44a |
Table 6 Copy number of ARGs in soybean soil with thioerythromycin residue
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±1.85a | 4.08±2.11a | 4.13±1.71a | 3.12±3.01a | 3.89±2.27a | 2.79±1.27a |
A2 | 5.60±0.85a | 5.36±0.61a | 4.57±0.75a | 5.75±0.31a | 5.61±0.56a | 3.71±0.33a | |
CK | 3.35±2.95a | 3.31±3.01a | 5.46±0.53a | 4.86±1.09a | 4.85±1.37a | 1.63±1.47a | |
结果期 Result period | A1 | 1.27±0.95a | 2.28±0.21a | 2.35±0.31a | 2.53±1.15a | 2.66±1.51a | 1.72±0.45a |
A2 | 1.34±0.75a | 2.18±0.42a | 2.52±0.23a | 3.65±0.17a | 3.81±0.43a | 1.64±0.51a | |
CK | 1.29±1.03a | 2.01±0.57a | 2.41±0.32a | 2.92±0.87a | 2.71±1.45a | 1.84±0.44a |
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