Different Functions of the Insect Soluble and Membrane-Bound Trehalase Genes in Chitin Biosynthesis Revealed by RNA Interference 英文参考文献.docVIP

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Different Functions of the Insect Soluble and Membrane-Bound Trehalase Genes in Chitin Biosynthesis Revealed by RNA Interference 英文参考文献.doc

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Different Functions of the Insect Soluble and Membrane-Bound Trehalase Genes in Chitin Biosynthesis Revealed by RNA Interference 英文参考文献

DifferentFunctionsoftheInsectSolubleandMembrane- BoundTrehalaseGenesinChitinBiosynthesisRevealed byRNAInterference JieChen1,BinTang1,2,HongxinChen1,QiongYao1,XiaofengHuang1,JingChen1,DaoweiZhang1, WenqingZhang1* 1StateKeyLaboratoryofBiocontrol,SchoolofLifeSciences,SunYat-senUniversity,Guangzhou,China,2HangzhouKeyLaboratoryofAnimalAdaptationandEvolution, HangzhouNormalUniversity,Hangzhou,Zhejiang,China Abstract Background:Trehalase,anenzymethathydrolyzestrehalosetoyieldtwoglucosemolecules,playsapivotalroleinvarious physiologicalprocesses.Inrecentyears,trehalaseproteinshavebeenpurifiedfromseveralinsectspeciesandaredivided into soluble (Tre-1) and membrane-bound (Tre-2) trehalases. However, no functions of the two trehalases in chitin biosynthesisininsectshaveyetbeenreported. Principal Findings: The membrane-bound trehalase of Spodoptera exigua (SeTre-2) was characterized in our laboratory previously. In this study, we cloned the soluble trehalase gene (SeTre-1) and investigated the tissue distribution and developmentalexpressionpatternofthetwotrehalasegenes.SeTre-1wasexpressedhighlyincuticleandMalpighiantubules, while SeTre-2 was expressed in tracheae and fat body. Inthe midgut, the two trehalase genes were expressed in different locations.Additionally,theexpressionprofilesofbothtrehalasemRNAsandtheirenzymeactivitiessuggestthattheymayplay differentrolesinchitinbiosynthesis.TheRNAinterference(RNAi)ofeitherSeTre-1orSeTre-2wasgene-specificandeffective,with efficiency rates up to 83% at 72 h post injection. After RNAi of SeTre-1 and SeTre-2, significant higher mortality rates were observed during the larva-pupa stage and pupa-adult stage, and the lethal phenotypes were classified and analyzed. Additionally,thechangetrendsofconcentrationoftrehaloseandglucoseappearedreciprocallyinRNAi-mutants.Moreover, knockdownofSeTre-1genelargelyinhibitedtheexpressionofchitinsynthasegeneA(CHSA)andreducedthechitincontentin thecuticletotwo-thirdsrelativetothecontrolinsects.Thechitinsyntha