溴氰菊酯降解菌的分离筛选与应用潜力评价
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摘要
为解决杀虫剂溴氰菊酯的残留污染问题及丰富其降解菌资源库,采用富集培养法,从长期被溴氰菊酯污染的土壤中分离筛选到1株高效降解菌株XQ08,经形态学特征、生理生化及16S rDNA鉴定其为嗜麦芽寡养单胞菌Stenotrophomonas maltophilia,并对菌株发酵条件、菌剂制备方法及应用潜力进行了初步研究。结果表明:在纯培养条件下,XQ08菌株5 d内对50 mg/L溴氰菊酯的降解率可达66.76%;在35℃、pH=8及以麦芽糖为碳源、氯化铵为氮源的条件下可获得理想的发酵效果,同时明确了XQ08液体菌剂的制备方法。分别在室内盆栽与田间种植的香菜上喷施高剂量(推荐剂量10倍,有效成分6.25 g/hm~2)的25 g/L溴氰菊酯乳油,于施药后2、24和48 h施用XQ08菌剂,3、5、7和9 d后,香菜上溴氰菊酯的降解率分别超过20%、50%、60%及70%;7 d后施用菌剂处理组溴氰菊酯残留量均低于0.5 mg/kg,9 d后残留量均低于0.2 mg/kg。本研究首次报道了嗜麦芽寡养单胞菌对溴氰菊酯的降解,发现其XQ08菌剂可有效促进溴氰菊酯的降解,且其降解符合一级动力学特征。嗜麦芽寡养单胞菌XQ08菌株能有效降低或清除蔬菜上的溴氰菊酯残留,具有较好的应用潜力和开发价值。
In order to solve the problem of residual contamination caused by the extensive use of deltamethrin and enrich the resource library of deltamethrin-degrading microorganisms, the effective deltamethrin-degrading XQ08 strain was isolated by enrichment culture method. The strain XQ08 was identified as Stenotrophomonas maltophilia based on the morphological, physiological and biochemical characteristics, and the 16 S rDNA homologue sequence analysis. The results showed that the strain XQ08 could degrade more than 66.76% of 50 mg/L deltamethrin in pure cultures within 5 d. Moreover,the ideal fermentation effect of the strain XQ08 could be achieved under the conditions of 35 ℃, pH8.0, maltose as a carbon source and ammonium chloride as a nitrogen source. And fermentation 24 h OD600 could reach 2.5. The optimal preparation technology of the liquid microbial agent of the strain XQ08 was developed. Under the condition of in door pot and field experiments, the XQ08 liquid microbial agent was sprayed on Coriandrum sativum at 2, 24 and 48 h after the spray of 25 g/L deltamethrin EC at a dose of 6.25 a.i.g/hm~2. The application of the liquid agent of the strain XQ08 could effectively degrade or remove deltamethrin residues on C. sativum. At 3, 5 and 7 d after the application of XQ08 microbial agent, the degradation rate of deltamethrin residues on C. sativum were more than20%, 50%, 60% and 70%, respectively. 7 and 9 d after the application of the liquid agent of the strain XQ08, the deltamethrin residues on the leaves of C. sativum were lower than 0.5 and 0.2 mg/kg,respectively. This is the first report on the degradation of deltamethrin by S. maltophilia. The degradation pattern of deltamethrin on C. sativum was found to follow first-order kinetics. The results presented here indicate that the strain XQ08 could effectively reduce or eliminate deltamethrin residues on vegetables. The strain XQ08 has attractive application potentia and value for further development.
In order to solve the problem of residual contamination caused by the extensive use of deltamethrin and enrich the resource library of deltamethrin-degrading microorganisms, the effective deltamethrin-degrading XQ08 strain was isolated by enrichment culture method. The strain XQ08 was identified as Stenotrophomonas maltophilia based on the morphological, physiological and biochemical characteristics, and the 16 S rDNA homologue sequence analysis. The results showed that the strain XQ08 could degrade more than 66.76% of 50 mg/L deltamethrin in pure cultures within 5 d. Moreover,the ideal fermentation effect of the strain XQ08 could be achieved under the conditions of 35 ℃, pH8.0, maltose as a carbon source and ammonium chloride as a nitrogen source. And fermentation 24 h OD600 could reach 2.5. The optimal preparation technology of the liquid microbial agent of the strain XQ08 was developed. Under the condition of in door pot and field experiments, the XQ08 liquid microbial agent was sprayed on Coriandrum sativum at 2, 24 and 48 h after the spray of 25 g/L deltamethrin EC at a dose of 6.25 a.i.g/hm~2. The application of the liquid agent of the strain XQ08 could effectively degrade or remove deltamethrin residues on C. sativum. At 3, 5 and 7 d after the application of XQ08 microbial agent, the degradation rate of deltamethrin residues on C. sativum were more than20%, 50%, 60% and 70%, respectively. 7 and 9 d after the application of the liquid agent of the strain XQ08, the deltamethrin residues on the leaves of C. sativum were lower than 0.5 and 0.2 mg/kg,respectively. This is the first report on the degradation of deltamethrin by S. maltophilia. The degradation pattern of deltamethrin on C. sativum was found to follow first-order kinetics. The results presented here indicate that the strain XQ08 could effectively reduce or eliminate deltamethrin residues on vegetables. The strain XQ08 has attractive application potentia and value for further development.
引文
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[2]陈宗懋,韩华琼,岳瑞芝.溴氰菊酯(DECIS)在茶叶中的残留降解[J].昆虫学报,1983,26(2):146-153.CHEN Z M,HAN H Q,YUE R Z.Degradation of pyrethroid DECIS(Nrdc 161)on tea-shoots[J].Acta Entomologica Sinica,1983,26(2):146-153.
[3]丁明,方学智,费学谦,等.柿果和土壤中菊酯类农药的残留检测和消解动态研究[J].江西农业大学学报,2006,28(4):512-515.DING M,FANG X Z,FEI X Q,et al.Quick determination and degradation of pyrethrins residuses in non-environmental pollution persimmon fruit and soil[J].Acta Agriculturae Universitatis Jiangxiensis,2006,28(4):512-515.
[4]李雪飞,杨艳刚,孙胜龙,等.蔬菜中有机磷、拟除虫菊酯类农药残留调查[J].环境与健康杂志,2006,23(5):418-420.LI X F,YANG Y G,SUN S L,et al.Analysis of organophosphorus and pyrethroid pesticides multi-residues in vegetables[J].J Environ Health,2006,23(5):418-420.
[5]刘永明,葛娜,崔宗岩,等.进口粮谷中农药残留状况普查与分析[J].食品安全质量检测学报,2016,7(3):1233-1239.LIU Y M,GE N,CUI Z Y,et al.Survey and analysis for pesticide residues in the imported grain[J].J Food Saf Qual,2016,7(3):1233-1239.
[6]张艳,苟春林,姜瑞,等.宁夏枸杞中拟除虫菊酯类农药残留风险评估[J].新疆农业科学,2015,52(12):2273-2278.ZHANG Y,GOU C L,JIANG R,et al.Risk assessment of pyrethroid pesticides in wolfberry of Ningxia[J].Xinjiang Agric Sci,2015,52(12):2273-2278.
[7]赵李娜,赖子尼,张威振,等.珠江河网水产品中菊酯类农药残留调查及健康风险评价[J].生态毒理学报,2014,9(3):437-444.ZHAO L N,LAI Z N,ZHANG W Z,et al.Residual levels and health risk assessment of pyrethroid pesticides in aquatic products of the pearl river delta[J].Asian J Ecotoxicol,2014,9(3):437-444.
[8]王文蝶,邓玉君,钱坤.溴氰菊酯农药在土壤中的残留·迁移研究[J].安徽农业科学,2011,39(21):12782-12783.WANG W D,DENG Y J,QIAN K.Study on residues and migration of deltamethrin in soil[J].J Anhui Agric,2011,39(21):12782-12783.
[9]郑伟华,赵建庄,马德英,等.溴氰菊酯的毒性和致突变性的研究进展[J].北京农学院学报,2004,19(1):77-80.ZHENG W H,ZHAO J Z,MA D Y,et al.Development of deltamethrin toxicity and mutagenicity[J].J Beijing Agric College,2004,19(1):77-80.
[10]ARSLAN H,ALTUN S,?ZDEMIR S.Acute toxication of deltamethrin results in activation of iNOS,8-OHdG and up-regulation of caspase 3,iNOS gene expression in common carp(Cyprinus carpio L.)[J].Aquat Toxicol,2017,187:90-99.
[11]李艳,张有林,王宏.农药残留降解方法研究[J].食品研究与开发,2004,25(6):31-33.LI Y,ZHANG Y L,WANG H.Studies on the methods of degrading pesticide residues[J].Food Res Dev,2004,25(6):31-33.
[12]钱玉琴,陈巧,董秀云,等.农药残留降解技术研究现状与展望[J].质量技术监督研究,2010(6):64-69.QIAN Y Q,CHEN Q,DONG X Y,et al.The current status and the prospect of the degradation of pesticide residues[J].Qual Tech Superv Res,2010(6):64-69.
[13]崔中利,崔利霞,黄彦,等.农药污染微生物降解研究及应用进展[J].南京农业大学学报,2012,35(5):93-102.CUI Z L,CUI L X,HUANG Y,et al.Advances and application of microbial degradation in pesticides pollution remediation[J].JNanjing Agric Univ,2012,35(5):93-102.
[14]BARRAGáN-HUERTA C,COSTA-PéREZ J,PERALTA-CRUZ J,et al.Biodegradation of organochlorine pesticides by bacteria grown in microniches of the porous structure of green bean coffee[J].Int Biodeterior Biodegrad,2007,59(3):239-244.
[15]WEN J P,LI H M,BAI J,et al.Biodegradation of 4-chlorophenol by Candida albicans PDY-07 under anaerobic conditions[J].Chin JChem Eng,2006,14(6):790-795.
[16]杨晓棠.溴氰菊酯降解菌的筛选及降解特性研究[D].长春:吉林农业大学,2008:1-30.YANG X T.Study on isolation and degrading characteristics of deltamethrin-degrading bacteria[D].Changchun:Jilin Agricultural University,2008:1-30.
[17]唐洁,张庆,曾朝懿,等.溴氰菊酯降解菌的分离与鉴定及其降解特性[J].西华大学学报(自然科学版),2014,33(3):108-112.TANG J,ZHANG Q,ZENG C Y,et al.Isolation and characterization of deltamethrin degrading strain and its degradation characteristic[J].J Xihua Univ(Nat Sci Ed),2014,33(3):108-112.
[18]MALONEY S E,MAULE A,SMITH A R.Microbial transformation of the pyrethroid insecticides:permethrin,deltamethrin,fastac,fenvalerate,and fluvalinate[J].Appl Environ Microbiol,1988,54(11):2874-2876.
[19]TALLUR P N,MEGADI V B,NINNEKAR H Z.Biodegradation of cypermethrin by Micrococcus sp.strain CPN 1[J].Biodegradation,2008,19(1):77-82.
[20]KALWASI?SKA A,K?SY J,WILK I,et al.Neustonic versus epiphytic bacteria of eutrophic lake and their biodegradation ability on deltamethrin[J].Biodegradation,2011,22(4):699-707.
[21]YANG Z H,JI G D.Quantitative response relationships between degradation rates and functional genes during the degradation of betacypermethrin in soil[J].J Hazard Mater,2015,299:719-724.
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