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1、中国水稻科学(ChinJRiceSci),2016,30(5):541-551http://www.ricesci.cnDOI:10.16819/j.1001G7216.2016.5198541稻曲病菌TGDNA插入突变体B2510的插入位点分析丁慧1,2俞咪娜2王亚会2于俊杰2尹小乐2薄惠文1,2黄星1刘永锋2,∗(1南京农业大学生命科学学院,南京210095;2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)MolecularCharacterizationofUstilaginoideavirensTGDNAInse
2、rtionMutantB2510DINGHui1,2,YUMiGna2,WANGYaGhui2,YUJunGjie2,YINXiaoGle2,BOHuiGwen1,2,HUANGXing1,LIUYongGfeng2,∗(1CollegeofLifeScience,NanjingAgriculturalUniversity,Nanjing210095,China;2InstituteofPlantProtection,JiangsuAcademyofAgriculturalSciences,Nanjing210014,China;∗Correspondingauthor,EGmail:
3、liuyf@jaas.acc.n)DINGHui,YUMina,YUJunjie,etal.MolecularcharacterizationofUstilaginoideavirensTGDNAinsertionmutantB2510.ChinJRiceSci,2016,30(5):541G551.Abstract:ToisolatethegeneconcernedwithmolecularpathogenicprocessofUstilaginoideavirens(U.virens),TGDNAintegrationflankingsequenceandthemutantgene
4、sofamutantstrainB2510wereanalyzed.ComparedwiththewildGtypeU.virensstrainP1,thepathogenicityofthemutantstrainB2510inthefieldwassignificantlydecreased.ThegrowthrateofB2510onMMmediumwasslowerthanthatofP1,butwasnotsignificantlydifferentonPSAandTB3mediumwhichwerenutritionallyendowed.ThemutantstrainB2
5、510didnotproduceconidiophoresinPSbrothmedium.GenomicSouthernboltanalysisconfirmedthatthereweredoubleTGDNAeventsinsertedingenomeofmutantstrainB2510.TheflankingU.virenssequencesofTGDNAobtainedbyTAILGPCRwereadjacentinthewildtypeandwithnosequenceslost,andonlyafewbasesofTGDNAwerechanged.TheTGDNAinser
6、tionsitewasinthepromoterregionofUV8b_1412anddownstream3′regionofUV8b_1386,respectively.RTGPCRanalysisconfirmedthattheexpressionofbothoftwogenesinB2510weresignificantlydecreased.ThegeneswhichwereaffectedbyTGDNAinsertionmaybeassociatedwithpathogenicityandparticipateintheregulationofpathogenicproce
7、ssofU.virensinrice.Keywords:Ustilaginoideavirens;TGDNAinsertionmutant;ATMTtransformation;pathogenicity;flankingsequence丁慧,俞咪娜,王亚会,等.稻曲病菌TGDNA插入突变体B2510的插入位点分析.中国水稻科学,2016,30(5):541G551.摘要:以稻曲病菌TGDNA插入突变体库中致病力减弱突变菌株B2510为材料,通