施氏假单胞菌YC-YH1对甲基对硫磷的降解及其代谢产物检测.pdf

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1、2015，34(11)：2097—2104农业环境科学学报2015年11月———————————————————————————————_．J矗urnalofAgro-EnvironmentScience●——————————————————————————————一施氏假单胞茵YC—YHl对甲基对硫磷的降解及其代谢产物检测史延华，任磊，贾阳，RuthNahurira，闫艳春+(中国农业科学院研究生院，北京100081)摘要：利用实验室已获取的菌株YC—YHl对甲基对硫磷进行降解，以高效液相色谱结合质谱分析(HP

2、LC—MS)测定菌株YC—YHl对甲基对硫磷的降解速率并检测其代谢产物，通过基因克隆研究菌株YC—YHl对甲基对硫磷降解的分子机制，同时分析代谢产物对菌株YC—YHl生长的影响。结果表明，菌株YC—YHl为施氏假单胞菌(Pseudomonasstutzeri)，能够高效降解甲基对硫磷，其同时含有甲基对硫磷水解酶基因mpd和有机磷水解酶基因ophC2。经质谱分析确定，菌株YC—YHl将甲基对硫磷水解为对硝基苯酚和二甲基硫代硫酸酯，其中对硝基苯酚显著抑制YC—YHl的生长。综上，施氏假单胞菌YC—YHl能够高效地降解

3、甲基对硫磷，但对硝基苯酚作为其代谢产物对YC—YH1的生长存在显著抑制作用。关键词：甲基对硫磷；对硝基苯酚；施氏假单胞菌；生物降解中图分类号：X171．5文献标志码：A文章编号：1672—2043(2015)11—2097—08doi：10．11654／jaes．2015．11．009BiodegradationofMethylParathionbyPseudomonasstutzeriYC-YHlandDetectionofItsMetabolitesSHIYan—hua，RENLei，JIAYang，RUTH

4、Nahurira，YANYan—chun+(GraduateSchoolofChineseAcademyofAgriculturalSciences，Beijing100081，China)Abstract：AnewlyisolatedbacterialstrainYC-YHl，whichcoulddegrademethyl-parathionefficiently，wasidentifiedaccordingto16SrRNAgeneanalysisincombinationwithphysiological，

5、biochemicalandBiologtests．Thepresentstudywasfocusedonitscapabilityofde-gradingmethylparathion．ThemetabolitesofmethylparathionwereidentifiedbyHPLC-MS．ChangesofmethylparathionanditsmetaboliteconcentrationsintheinorganicmediumweredeterminedbyHPLC．Theprimersofgen

6、esinvolvedinthemethylparathiondegradationweredesignedandthetargetfragmentswereobtainedbygenecloning．TheinfluenceofdegradationintermediatesonthegrowthofthestrainYC-YHlwasexaminedusingfilterpaperandplatemethods．ThecolonicandmicroscopicmorphologiesofthestrainYC-

7、YHlwasdetermined．The16SrRNAgenesequenceshowedthatYC—YHlhad98％similaritytoPseudomonasstutzeri．PseudomonasstutzeriYC-YHlshowedhighdegradationefficiencyforparathion—methyl．p-nitrophenolanddimethylphosphorothioatewereidentifiedastheintermediates．p-hi-trophenolcon

8、centrationsincreasedalongwithdecreaseofmethylparathionconcentrationsintheinorganicmedium．However，p-nitrophenolsignificantlyinhibitedthegrowthofthestrainYC—YH1intheinorganicmedium．Methylpa