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1、中国水稻科学(ChinJRiceSci),２０１５,２９(３):３１１－３１８http://www．ricesci．cnDOI:１０．３９６９/ji．ssn．１００１Ｇ７２１６．２０１５．０３．０１１３１１稻曲病菌TＧDNA插入突变体B１４６４插入位点分析王亚会刘永锋∗陆凡俞咪娜黄磊郑梦婷于俊杰尹小乐(江苏省农业科学院植物保护研究所,南京２１００１４;∗通讯联系人,EＧmail:liuyf＠jaas．ac．cn)MolecularCharacterizationofTＧDNAIntegrationintoUstilaginoideavirensMutantB１４
2、６４WANGYaＧhui,LIUYongＧfeng∗,LUFan,YUMiＧna,HUANGLei,ZHENGMengＧting,YUJunＧjie,YINXiaoＧle(InstituteofPlantProtection,JiangsuAcademyofAgriculturalSciences,Nanjing２１００１４,China;∗Correspondingauthor,EＧmail:liuyf＠jaas．acc．n)WANGYahui,LIUYongfeng,LUFan,etal．MolecularcharacterizationofTＧDNAinte
3、grationintoUstilaginoideavirensmutantB１４６４．ChinJRiceSci,２０１５,２９(３):３１１Ｇ３１８．Abstract:WithanavirulentUstilaginoideavirensstrainB１４６４asmaterial,weanalyzedthechangesinbiologicalmorphologyandpathogenicmechanism．ComparedtothewildＧtypeU．virensstrainP１,themutantstrainB１４６４showednopathogenici
4、tyinthefield．TherateofgrowthonMMmediumwasnotdifferenttoP１andB１４６４．OnPSAandTB３mediumtheratesofgrowthwerenutritionallyendowed．B１４６４grewslowerandproducedfewconidiophoresinPSmedium,anditsconidiophoresmorphologyandsporesizebecamesmaller．GenomicSouthernboltanalysisconfirmedthatB１４６４wassing
5、leTＧDNAinsertionalevents．TheflankingsequencesofTＧDNAobtainedbyTAILＧPCRwerenonＧadjacentinthewildtype．Itwasfoundsthatthemutantstrainlostabout２０kbsequencesintheinsertionsitecomparingwiththewildtypestrain．TheexpressionoftheflankinggenesoftheTＧDNAwasdownＧregulated,detectingbysemiＧquantita
6、tiveRTＧPCR．ThereasonofthechromosomerearrangementandabnormalexpressionofsomegenesistheinsertionofTＧDNA,probably,leadingtothemutantstrainB１４６４lostitspathogenicity．Keywords:Ustilaginoideavirens;TＧDNAinsertionmutant;pathogenicity;malatesynthase;coppertransporter王亚会,刘永锋,陆凡,等．稻曲病菌TＧDNA插入突变
7、体B１４６４插入位点分析．中国水稻科学,２０１５,２９(３):３１１Ｇ３１８．摘要:以稻曲病菌致病力丧失的突变菌株B１４６４为材料,对其生物学形态和分子遗传变异进行分析.B１４６４田间接种表现为不致病,与野生型菌株相比,在营养贫瘠的MM培养基上生长速率没有显著差异,而在PSA和TB３培养基中生长速率较慢,并且在PS液体培养基中产孢能力下降,同时突变菌株的分生孢子与野生型相比形态和大小均发生变化.Southern杂交结果显示TＧDNA在突变菌株B１４６４中以单拷贝形式插入,利用TAILＧPCR技术扩增得到的紧邻TＧDNA两侧的侧翼序列在野生型中不相邻,与野生型菌株
8、基因组比对发现突变菌株插
