中国蔬菜2011(8):76-80CHINAVEGETABLES黄瓜棒孢叶斑病(褐斑病)病菌细胞壁降解酶产生条件及活性分析刘志恒亓烨黄欣阳杨红侯悦张睿(沈阳农业大学植物保护学院,辽宁沈阳110161)摘要:通过对活体外细胞壁降解酶产生条件、活体接种黄瓜棒孢叶斑病菌后细胞壁降解酶活性变化及活体内外产生的细胞壁降解酶活性比较的研究,结果表明:多聚半乳糖醛酸酶(PG)、果胶甲基半乳糖醛酸酶(PMG)、多聚半乳糖醛酸反式消除酶(PGTE)、果胶甲基反式消除酶(PMTE)、羧甲基纤维素酶(Cx)和β-葡萄糖苷酶6种酶均有特定的最适培养条件,持续振荡培养利于各种细胞壁降解酶的产生;Cx和β-葡萄糖苷酶在培养9d时活性达到高峰,PG和PMG在培养12d时活性达到高峰,PGTE、PMTE在培养15d时活性最高;PG和PMG产生的最适温度为25℃,PGTE、PMTE、Cx、β-葡萄糖苷酶产生的最适温度为28℃;PG、PMG、Cx和β-葡萄糖苷酶的活性在pH为6时最高,PGTE和PMTE的活性在pH为8时达到最高峰。4种果胶酶的活性在接种后第3天活性最高;而两种纤维素酶在接种后第5天活性最高。病菌在活体内和活体外产生的细胞壁降解酶的活性明显不同。关键词:黄瓜棒孢叶斑病(褐斑病);多主瓜棒孢霉;细胞壁降解酶;活性中图分类号:S436.421.1+9文献标识码:A文章编号:1000-6346(2011)08-0076-05ConditionsandActivityAnalysisofCellWallDegradingEnzymesProducedfromCorynesporacassiicolaofBrownSpotofCucumberLIUZhi-heng,QIYe,HUANGXin-yang,YANGHong,HOUYue,ZHANGRui(CollegeofPlantProtection,ShenyangAgriculturalUniversity,Shenyang110161,Liaoning,China)Abstract:Aseriesofcellwalldegradingenzymes(CWDEs)includingpolygalacturonase(PG),polymethyl-galacturonase(PMG),polygalacturonicacidtrans-eliminase(PGTE),pectinmethyl-trans-eliminase(PMTE)carboxymethylcellulase(Cx)andβ-glucosidasewerefoundbothinvitroandvivofromCorynesporacassiicolaofbrownspotofcucumber.Basedonthecellwalldegradingenzymesproducedinvitroconditions,invivoafterinoculationofbacteriacellwall-degradingenzymesandinvivoproducedoutsidethecellwall-degradingenzymeactivitycomparisonstudy,theresultsshowthateverykindofcellwalldegradingenzymehasaspecificoptimalcultureconditions.Shakeculturemakemorefortheproductionofcell-walldegradationcomparedtothestationaryculture.TheactivitiesofCxandβ-glucosidasereachedapeakwhencultured9d,PGandPMGreachedapeakwhencultured12d,whilePGTEandPMTEreachedapeakwhencultured15d.PGandPMGshowedmaximumactivitiesat25℃culturetemperature.PGTE,PMTE,Cxandβ-glucosidaseshowedmaximumactivitiesat28℃culturetemperature.PG,PMG,Cxandβ-glucosidaseshowedmaximumactivitiesatculturepH6.PGTEandPMTEshowedmaximumactivitiesatculturepH8.Fourkindsofpectinaseshowedmaximum收稿日期:2010-11-06;接受日期:2011-01-06作者简介:刘志恒,男,教授,主要从事植物病理学、真菌学等教学与研究工作,E-mail:lzhh1954@163.com《中国蔬菜》学术论文下载www.cnveg.org2011(8)刘志恒等:黄瓜棒孢叶斑病(褐斑病)病菌细胞壁降解酶产生条件及活性分析77activitiesinthethirddayafterinoculation.Twokindsofcellulaseshowedmaximumactivitiesinthefifthdayafterinoculation.Theactivityofcellwalldegradingenzymesinvivoandinvitrowassignificantlydifferent.Keywords:Cucumbertargetleafspot(brownspot);Corynesporacassiicola;Cellwalldegradingenzymes;Activity黄瓜棒孢叶斑病(褐斑病)是由多主棒孢霉〔Corynesporacassiicola(Berk&Curt)Wei.〕引起的叶斑类病害(李宝聚等,2008;李长松等,2009),是黄瓜生产上的重要病害之一。该病于1992年在辽宁省鞍山海城首先发现(房德纯和傅俊范,1994),其后相继在沈阳、大连瓦房店以及河北青县等地发生(李宝聚等,2008)。近年来,该病在辽宁省的发生及危害有上升趋势,在辽宁南部、辽宁西部和辽宁东部...