蘇云金芽孢桿菌晶體殺蟲蛋白Cry1Ac基因在蟲生真菌球孢白僵菌中的表達.pdf

1、安徽農業(yè)大學碩士學位論文蘇云金芽孢桿菌晶體殺蟲蛋白Cry1Ac基因在蟲生真菌球孢白僵勞中的表達姓名：劉振邦申請學位級別：碩士專業(yè)：森林保護學指導教師：李增智黃勃201106IIAbstractFungalinsecticidesisastofenvironmentalfriendlyenvironmentallysafebiopesticidewithgoodsafetytohumanvertebratesnontargetinvert

2、ebratessubstantialsustainablecontroleffecteasinessofmassproductionwithartificialmedia.Ontheotherhtheyaresubjectedeasilytoenvironmentalfactskillinsectsslowlynotsteadilywhichhasconstrainedthedevelopmentoftheuseofthefungi.M

3、oleculerpathogenicmechanismshavebeenstudiedindertothevirulentgeneswhichcanbeimprovedbygeneengineering.Bacillusthuringiensisisgrampositivebacteriumoneofkindswhichcanfmparaspalcrystalprotein(insecticidalcrystalproteinICP)δ

4、endotoxininthestabilityofthebacteriagrowth.Itisstrongpoisonousfcespeedingdeadlyfpeststhatisavirulententomopathogenfthenaturalenemyofpests.Cry1AcgeneofB.thuringiensisisanimptantvirulentgenebutitisunknownwhetheritcanenhanc

5、ethevirulenceofafungus.InthepresentstudytheexogenousCry1AcwastransferredintotwowildtypeisolatesofBeauveriabassianaonefromtheMasson’rpillerDendrolimuspunctatusanotherfromtheAsiancnberOstriniafurcanalis，toconstitutetwogene

6、engineeredstrainswithonecopiedoverexpressedCry1Ac.AvectofpbarGPE1Cry1AccontainingtheingmarkerbargeneexogenousgeneCry1AcwasconstoitutedthentransferredintoRCEF0013RCEF3383byblastospetranfmmethod.Transfmantswerederivedthrou

7、ghsievingonSDAYmediumcontaining100?gmlglufosinateammoniumresultinginthemonovalentCry1ActransgenicengineeredstrainsRCEF0013G1RCEF3383G1.TheCry1Acgenewasoverexpressedwiththeengineeredstrains.TheresultofbioassayfDendrolimus

8、punctatusshowedthatascomparedtothewildtypeisolateRCEF0013theLC25ofRCEF0013G1tothepinecaterpillarsdecreasedby5.4foldstheLD25reducedby7.1timesLT50shtenedby2.2dindicatingasubstantialvirulenceenhancement.Comparedtospraytreat

9、mentthecumulativemtalityoffeedingtreatmentfeedingwithspraytreatmentwereincreasedby55.6%63.0%LT50shtenedby4.5ddays5.3ddays.ThesedataindicatedthatthestomachtoxicityofBtdeltaendotoxingeneenhancedthevirulenceofB.bassianagrea