《农杆菌介导的AtMYB118基因转化橡胶树易碎胚性愈伤组织体系优化》.pdfVIP

南方农业学报 JournalofSouthernAgriculture 2014．45(7)：1137—1146 ISSN2095—1191；CODEN NNXAAB — http：／／www．nfnyxb．cn DOI：10．3969／j：issn．2095-1191．2014．7．1137 Optimizationofsystem forAgrobacterium tumefaciens mediatedAtM YBll8genetictransformation intofriable eemmbOrryyoo~genicccaalllliiinHHeveabbrraassiillJieemnsm BIZheng-hong，LIZhe ，WANGZhi—qiu，DAIXue—mei，FANGJia—lin，HUANG Hua—sun，LINW ei-fu ，LARDETLudovic，MONTOROPascal。，CARRONMarc—Philippe3， LECLERCQJulie。，ZUOJian-ru，MUJin-ye，YANGXiao—hui，LIXiang (CollegeofAgronomy，HainanUniversity，Haikou570228，China；RubberResearchInstitute，CATASK／eyLaboratoryof RubberBiologyandGeneticResourcesUtilization，Ministry ofAgriculture／StateEngineeringandTechnologyResearchCenter forKeyTropicalCrops，Danzhou，Hainan 571737，China；EquipeBURST-UMRAGAP，D6partementBIOS，CIRAD， Montpellier34398，France；InstituteofGeneticsandDevelopmentalBiology，CAS，Beijing100101，China；Environment andPlantProtectionCollege，HainanUniversity，Haikou570228，China) Abstract：【Objective】AnorthogonaldesignL25(5)wasused to optimize Agrobaetedum—mediated AtMYBI18 genegenetictransformation system offriableembryogeniccalliin Heveabrasiliensisinordertoprovidereferencesfor geneticimprovementofitsclones． 【Method】Kanamycinataconcentrationof75．0mg／Lwasusedtoselecttrans— formed calli．Theeffectsofsixfactors ．，precuhuretime，Agrobacterium growthphase (OD600)，acetosyringone (AS)concentration，infection time．co-culturetemperatureand co-culture timeongenetictransofrmationwere eval— uatedbyGUStransientexpressiondetection． 【Result】Sixfactorsweref0undtosignificantlyaffecttransformationfre— quencyoflong-term culturalfriableembrvogenic calli．Thehighesttransofrmation efficiency wasachieved by 0一day precuhure．1ong—term cuhuralfriabl