Genetic and Proteomic Characterization of rpoB Mutations and Their Effect on Nematicidal Activity in Photorhabdus luminescens LN2 英文参考文献.docVIP

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Genetic and Proteomic Characterization of rpoB Mutations and Their Effect on Nematicidal Activity in Photorhabdus luminescens LN2 英文参考文献.doc

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Genetic and Proteomic Characterization of rpoB Mutations and Their Effect on Nematicidal Activity in Photorhabdus luminescens LN2 英文参考文献

GeneticandProteomicCharacterizationofrpoB MutationsandTheirEffectonNematicidalActivityin PhotorhabdusluminescensLN2 XuehongQiu,XunYan,MingxingLiu,RichouHan* GuangdongEntomologicalInstitute,Guangzhou,Guangdong,China Abstract Rifampin resistant (RifR) mutants of the insect pathogenic bacterium Photorhabdus luminescens LN2 from entomopatho- genic nematode Heterorhabditis indica LN2 were genetically and proteomically characterized. The RifR mutants showed typical phase one characters of Photorhabdus bacteria, and insecticidal activity against Galleria mellonella larvae, but surprisingly influenced their nematicidal activity against axenic infective juveniles (IJs) of H. bacteriophora H06, an incompatiblenematodehost. 13outof34RifR mutantslost theirnematicidalactivityagainstH06IJsbutsupported the reproductionofH06nematodes.7nematicidal-producingand7non-nematicidal-producingRifRmutantswererespectively selectedforrpoBsequenceanalysis.rpoBmutationswerefoundinall14RifRmutants.TherpoB(P564L)mutationwasfound inall7mutantswhichproducednematicidalactivityagainstH06nematodes,butnotinthemutantswhichsupportedH06 nematodeproduction.AllelicexchangeassaysconfirmedthattheRif-resistanceandtheimpactonnematicidalactivityof LN2bacteriawereconferredbyrpoBmutation(s).Thenon-nematicidal-producingRifRmutantwasunabletocolonizeinthe intestinesofH06IJs,butabletocolonizeintheintestinesofitsindigenousLN2IJs.Proteomicanalysisrevealeddifferent protein expression between wild-type strain and RifR mutants, or between nematicidal-producing and non nematicidal- producing mutants. At least 7 putative proteins including DsbA, HlpA, RhlE, RplC, NamB (a protein from T3SS), and 2 hypotheticalproteins(similartounknownproteinYgdHandYggEofEscherichiacolirespectively)wereprobablyinvolvedin the nematicidal activity of LN2 bacteria against H06 nematodes. This hypothesis was further confirmed by creating insertion-deletion mutants of three selected corresponding genes (the downregulated rhlE and namB, and u