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1、农业生物技术学报,2010年,第18卷,第5期,第827~835页JournalofAgriculturalBiotechnology,2010,Vol.18,No.5,827~835研究论文AnArticle 梨火疫菌(Erwiniaamylovora)环腺苷酸受体蛋白基因(crp)的功能分析*刘倩倩于洋洋宋俊贤胡白石范加勤刘凤权南京农业大学农业部病虫监测与治理重点开放实验室,南京210095*通讯作者,hbs@njau.edu.cn摘要梨火疫菌(Erwiniaamylovora)可引起梨、苹果
2、等蔷薇科(Rosaceae)植物的火疫病。在菊欧文氏菌(Erwiniachrysanthemi)中,由crp基因编码的环腺苷酸受体蛋白(cyclicadenosinemonphosphate(cAMP)receptorprotein,CRP)对果胶酶基因的表达调控和菌株致病性起着重要的作用。本研究首次鉴定并克隆出梨火疫菌中的crp同源基因,命名为Eacrp,并通过同源重组的方法,构建了梨火疫菌的crp基因突变体Ea驻crp以及互补子,进行了致病性、过敏性反应、胞外多糖、鞭毛运动等一系列相关表型的鉴
3、定。结果表明,crp基因影响着梨火疫菌的致病性、胞外多糖、游动性、生长情况等多种生物学特性,然而,Ea驻crp仍能引起烟草过敏性反应,并且在过氧化氢敏感度以及沉降性、生物膜和AI2 信号分子的生成方面与野生型菌株相比差异明显。本研究结果说明,梨火疫菌crp基因对病菌的胞外多糖分泌、生长、游动性以及致病性方面具有关键作用。关键词梨火疫菌,crp基因,致病性,胞外多糖FunctionalCharacterizationofCyclicAdenosineMonphosphate (cAMP) Recpt
4、orProteinGene(crp)fromErwiniaamylovora*LiuQianqianYuYangyangSongJunxianHuBaishi FanJiaqinLiuFengquan KeyLaboratoryofMonitoringandManagementofPlantDiseasesandInsects,MinistryofAgriculture,NanjingAgricultureUniversity, Nanjing210095,China*Correspondinga
5、uthor,hbs@njau.edu.cnDOI:10.3969/j.issn.16747968.2010.05.001AbstractErwiniaamylovoracausesfireblightofpear,apple,andmanyoftheRosaceaefamily.Thecyclicadenosinemonphosphate(cAMP)receptorprotein(CRP),whichisencodedbycrpgene,playsacrucialrolein expression
6、ofthepectinolysisgenesandregulationofthepathogenicityinErwiniachrysanthemi.Inthisstudy,the homologofcrpgene,designatedEacrp,wasidentifiedandclonedinErwiniaamylovora.Acrpdisruptionmutant Ea驻crpwassuccessfullygeneratedbyhomologousrecombination.Complemen
7、tstrainEa驻crp(pMEcrp)wasalso constructed.Thencrpgenewascharacterizedtovirulence,hypersensitiveresponse(HR),extracellularpolysaccharide(EPS),motilityandotherphenotypesofthephytopathogensErwiniaamylovora.Theresults showed that thecrpmutantEa驻crpdisplaye
8、dthereducedvirulence,EPSproductionandgrowthinvitroand motility. However,comparedtowildtypestrainNCPPB1665(Ea1665),theinductionofHRinnonhosttobacco,peroxide resistance,sedimentation,biofilmsynthesisandAI2productionofthecrpmutantEa驻crpwasnot aff