大蒜病毒病原鑒定及其檢測技術(shù)研究.pdf_第1頁
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1、黑龍江八一農(nóng)墾大學(xué)碩士學(xué)位論文大蒜病毒病原鑒定及其檢測技術(shù)研究姓名：張威申請學(xué)位級別：碩士專業(yè)：植物病理學(xué)指導(dǎo)教師：張勻華李學(xué)湛20070401AbstractThegwficinHeiLongJiangwaswidelyinfectedbycomplexviruses，throughinvestigatingNaturalinfectedgarliccropsexhibiteddiversiformsymptomssuchasmasa

2、ic、crimple、leafroll、abnormality、eliolation、putrescenceandsoonInmyexperiment，thevirusesinfectinggarlicsweredetectedandidentifiedbyserology、bioassay、electronmicroscope、molecularbiologicaltechniquesThroughbioassay，indicator

3、hostswereinoculatedbygarlicviruses，thegarlicvirusepathogenyswereelementarilyidentifiedThebiologicassayvalidatedthatAlliwnporrwn、Alliwncepa、Hcia丘蚰andGomphrenaglobosawasbetterindicatorhostofLYSV、OYDV、GCLVandPVXrespectively

4、VmxsparticleswereobservedinpurifiedsapfrominfectedgarlicorinfectedindicatorhostTwokindsofflexousfilamentousparticleswereobserved，theonewasabout3001000nmlongtheotherwasabout18502000nmlongVhusparticleswereobservedbyelectro

5、nmicroscopyullrathins(clion。thereweretwokindsofrimsparticles，theonespecieswasfilamentous，theotherwasrhabdovimsesTheyexistincytoplasmandformedvariousinclusionbodysLongfilamentousandrhabdovimseswerefirstfoundingarlicThelea

6、vesweredetectedbyDASELISA，andtheresultshowedthatmasaicgarliccontainedLYSVandPVX，etiolationgarliccxmlainedOYDVandGCLV，leafrollgarliccontainedGCLV、LYSVandOYDVThereinto，PVXwerefirstdeletedingarlicTheserologicalassayfor37gar

7、licfieldsamplesinHeiLongJiangshowedthattheinfectionrateofLYSV、GCLV、OYDV、ShLVandPVXwas95％、86％、74％、50％and78％respectivelyandmostnfthemwerecomplexinfectedWiththespecificprimerswhichweredesignedbasedontheGarliccommonlatentvir

8、us(GCLV)comproteinconservedsequence(withoptimizLngthedeteclionsystem)onegonefragment(300t)p)xTqasamplifiedbyreversetranscriptionpolymerasechainreactinn(RTPCR)usingthetotalRNAoftheplantinfectedbyGCLVThegenewaschonedandseq

9、uencedTheresultwascomparedwiththesequenceofotherisolatesofGCLVinGenBankTheresultshowedthatthehighesthomologycouldreach98％ofnucleicacidand97％ofaminoacidThedetectingspecificationandsemilivitywereverifiedWahthespecificprime

10、rswhichweredesignedbasedootheLeekyeUowstripevims(cYsV)nucleicaddconservedsequence，onegonefragment(1032hp)wasamplifiedbyRTPCRusingthetotalRNAoftheplantinfectedbyLYSVThegenewaschonedandsequenced，theCPsequencewasincludedThe

11、sequenceofHIAwascomparedwimthesequenceoftheother23isolatesofLYSVtheresultshowedthatthehighesthomologycouldreach88覦ofnucleicacidand931％ofaminoacidwhichprovedmatthevimswasLYSVTheCPaminoacidsequencesofdifferentisolatesofLYS

12、Vwerecomparedtheresultsbowed：theseweregreatdifferencesontheaminoacidsequenceamongtheLYSVisolates，thehomolo盟，rangeedfrom785～100％ThephylogeniclrceofLYSVwasclusteredintofourgroupsTheisolateofHLJwasinthesa／Ilegroupwiththeiso

13、latesofSouthKoreaandJapan，theyrepresentedregionallycorrelationWiththespecificprimerswhichweredesignedbasedonthePotatovirusx(Pvx)cP，thegenefragment(714bp)wasrespectivelyamplifiedbyRTPCRwiththesamereactedconditionusingthet

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