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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