Screening Identification and Degradation Characteristics of Three Iprodione-degrading Strains

doi:10.13560/j.cnki.biotech.bull.1985.2022-1372

Abstract

Abstract:

To isolate the iprodione-degrading microbial resources suitable for Qinghai-Tibet Plateau, three efficient iprodione-degrading strain Y-20, Y-29 and Y-32 were isolated from the greenhouse soil in Tibet by enrichment culture method. These strains were preliminarily identified based on the morphological characteristics, physiological and biochemical characteristics, and 16S rRNA gene sequence analysis. The effects of NaCl concentration（m/V）, pH value, temperature, inoculation amount（V/V）and initial iprodione concentration on the iprodione-degradation rates of these strains were also investigated. The metabolic pathways of iprodione and ipaH gene in these strains were analyzed using gas chromatography method and polymerase chain reaction technique, respectively. The results showed that strain Y-20, Y-29 and Y-32 were identified as three species of Microbacterium genus. Under the optimal iprodione-degrading condition of 1.0% NaCl（m/V）, pH 7.0, 25-30℃, 5% inoculation amount（V/V）and 100 mg/L initial iprodione concentration, 100 mg/L iprodione was completely degraded by these strains within 8-12 h. These strains decomposed iprodione into N-（3,5-dichlorophenyl）-2,4-dioximidazolidine and isopropyl carbamate, and had a highly similar ipaH gene（99.14%-99.69%）. This study provides the strain resources and theoretical basis for the bioremediation of iprodione-contaminated environment at high altitude area.

Key words: iprodione, degradation, enrichment culture, Microbacterium, metabolic pathway, ipaH gene, gas chromatography

PAN Hu, ZHOU Zi-qiong, TIAN Yun. Screening Identification and Degradation Characteristics of Three Iprodione-degrading Strains[J]. Biotechnology Bulletin, 2023, 39(6): 298-307.

Figures/Tables 9

References 26

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测试项目Test item	Y-20	Y-29	Y-32	测试项目Test item	Y-20	Y-29	Y-32
邻硝基苯-半乳糖苷2-Nitrophenyl-galactopyranoside	+	+	-	Kohn明胶Kohn gelatin	+	+	-
精氨酸Arginine	-	-	-	葡萄糖Glucose	+	+	-
赖氨酸Lysine	-	-	-	甘露醇Mannitol	+	+	-
鸟氨酸Ornithine	-	-	-	肌醇Inositol	-	-	-
柠檬酸钠Sodium citrate	+	+^w	+	山梨醇Sorbitol	-	-	-
硫代硫酸钠Sodium thiosulfate	-	-	-	鼠李糖Rhamnose monohydrate	+	-	-
尿素Urea	-	-	-	蔗糖Sucrose	+	+	-
色氨酸Tryptophan（TDA）	+	+^w	+	密二糖Disaccharide	+	-	-
色氨酸Tryptophan（IND）	-	-	-	苦杏仁苷Amygdalin	+	+	-
丙酮酸盐Pyruvate	+	+	-	阿拉伯糖Arabinose	+	-	+^w

测试项目Test item	Y-20	Y-29	Y-32	测试项目Test item	Y-20	Y-29	Y-32
邻硝基苯-半乳糖苷2-Nitrophenyl-galactopyranoside	+	+	-	Kohn明胶Kohn gelatin	+	+	-
精氨酸Arginine	-	-	-	葡萄糖Glucose	+	+	-
赖氨酸Lysine	-	-	-	甘露醇Mannitol	+	+	-
鸟氨酸Ornithine	-	-	-	肌醇Inositol	-	-	-
柠檬酸钠Sodium citrate	+	+^w	+	山梨醇Sorbitol	-	-	-
硫代硫酸钠Sodium thiosulfate	-	-	-	鼠李糖Rhamnose monohydrate	+	-	-
尿素Urea	-	-	-	蔗糖Sucrose	+	+	-
色氨酸Tryptophan（TDA）	+	+^w	+	密二糖Disaccharide	+	-	-
色氨酸Tryptophan（IND）	-	-	-	苦杏仁苷Amygdalin	+	+	-
丙酮酸盐Pyruvate	+	+	-	阿拉伯糖Arabinose	+	-	+^w

样品来源 Source	分离菌株种类 Species of isolates	异菌脲初始浓度 Initial concentration of iprodione	降解率 Degradation rate/%	降解时间 Degradation time	降解基因 Degradation genes
土壤 Soil	Arthrobacter sp. MA6^[13]	9.90 mg/L	86.7	7 d	ND
土壤 Soil	Pseudomonas fluorescens, Pseudomonas sp., Pseudomonas paucimobilis^[14]	8.25 mg/L	100	20-24 h	ND
土壤 Soil	Zygosaccharomyces rouxii DBVPG 6399^[24]	1 mg/L	100	9 d	ND
农田土壤 Farmland soil	Arthrobacter sp.CQH^[15]	100 mg/L	100	112 h	ND
土壤 Soil	Microbacterium sp. CHQ-1^[12]	100 mg/L	100	96 h	ND
活性污泥 Activated sludge	Microbacterium sp.YJN-G^[17,25]	100 mg/L	100	24 h	ipaH
酸性土壤 Acidic soil	Arthrobacter sp. C1, Achromobacter sp. C^2[16]	60 mmol/L	100	10 d	ND
三七根际土壤 Root-zone soil of Panax notoginseng	Bacillus sp.KMS-1^[18]	25 mg/L	41.4	7 d	ND
葡萄园种植土壤 Vineyards grow soil	Paenarthrobacter sp. YJN-5^[8,25] Paenarthrobacter sp. YJN-D	1.5 mmol/L	95	80 h	ipaH, ddaH, duaH
大棚土壤 Greenhouse soil	Microbacterium sp. Y-20、Y-29、Y-32	100 mg/L	100	8-12 h	ipaH

样品来源 Source	分离菌株种类 Species of isolates	异菌脲初始浓度 Initial concentration of iprodione	降解率 Degradation rate/%	降解时间 Degradation time	降解基因 Degradation genes
土壤 Soil	Arthrobacter sp. MA6^[13]	9.90 mg/L	86.7	7 d	ND
土壤 Soil	Pseudomonas fluorescens, Pseudomonas sp., Pseudomonas paucimobilis^[14]	8.25 mg/L	100	20-24 h	ND
土壤 Soil	Zygosaccharomyces rouxii DBVPG 6399^[24]	1 mg/L	100	9 d	ND
农田土壤 Farmland soil	Arthrobacter sp.CQH^[15]	100 mg/L	100	112 h	ND
土壤 Soil	Microbacterium sp. CHQ-1^[12]	100 mg/L	100	96 h	ND
活性污泥 Activated sludge	Microbacterium sp.YJN-G^[17,25]	100 mg/L	100	24 h	ipaH
酸性土壤 Acidic soil	Arthrobacter sp. C1, Achromobacter sp. C^2[16]	60 mmol/L	100	10 d	ND
三七根际土壤 Root-zone soil of Panax notoginseng	Bacillus sp.KMS-1^[18]	25 mg/L	41.4	7 d	ND
葡萄园种植土壤 Vineyards grow soil	Paenarthrobacter sp. YJN-5^[8,25] Paenarthrobacter sp. YJN-D	1.5 mmol/L	95	80 h	ipaH, ddaH, duaH
大棚土壤 Greenhouse soil	Microbacterium sp. Y-20、Y-29、Y-32	100 mg/L	100	8-12 h	ipaH