中文摘要：

为了优化根癌农杆菌介导的葡萄愈伤组织瞬时转化体系,该研究以欧洲葡萄品种无核白（Vitis vinifera L.cv.Thompson Seedless）单芽茎段诱导的愈伤组织为材料,探讨重悬液pH、菌液浓度、真空渗透时间等主要因素对葡萄愈伤组织瞬时转化效率的影响。结果表明：（1）以激素组合分别为1.0mg/L BAP、1.0mg/L BAP＋0.02mg/L NAA、2.0mg/L BAP＋0.02mg/L NAA和4.0mg/L BAP＋0.02mg/L NAA的系列培养基更适合无核白葡萄单芽茎段逐步诱导胚性愈伤组织。（2）葡萄愈伤组织瞬时转化体系中,重悬液pH 5.1,菌液浓度OD6001.0,真空渗透20min为转化效率最佳条件。（3）利用优化的瞬时转化体系瞬时转化无核白葡萄的不同组织,发现在不同器官中转化效率存在显著差异。其中以愈伤组织为受体的转化效率显著高于其他器官（65 231.99±3 339.29mU/g）,而且愈伤组织的GUS组织化学染色最深,以叶片为受体的转化效率则最低。利用该体系转化质粒载体pCAMBIA0390∷GUS,瞬时表达产物经过GUS蛋白活性检测,结果表明该研究优化的葡萄愈伤组织瞬时转化体系有助于外源基因在葡萄愈伤组织内的表达,为后期通过转基因技术研究目标基因功能奠定了技术基础。

英文摘要：

In order to improve the Agrobacterium tumefaciens-mediated transient expression system in grapevine callus,we used the callus of grape ‘Thompson Seedless＇,which was induced from stem segment with single bud,to analyze some major factors influencing the transient expression efficiency,such as the pH of A.tumefaciens suspension,the concentration of A.tumefaciens suspension and the time of vacuum infiltration.The results showed that：（1）the best media for the induction of grapevine callus were hormone combination series of 1.0mg/L BAP,1.0mg/L BAP＋0.02mg/L NAA,2.0mg/L BAP＋0.02mg/L NAA and 4.0mg/L BAP＋0.02mg/L NAA,comparing with the hormone combination which is added0.1mg/L 2,4-D or 0.1mg/L TDZ.（2）The optimal combination was that the pH of A.tumefaciens suspension,the OD_（600） of A.tumefaciens suspension and the time of vacuum infiltration were 5.1,1.0and 20 min,respectively.（3）The optimized transient expression system had significant influence on the transient expression efficiency in different organs of Thompson Seedless.The transient expression efficiency of callus is the significantly highest（6 5231.99±3 339.29mU/g）,and the GUS staining result of callus is the darkest.However,the transient expression efficiency of leaves is the lowest.The transient expression vector pCAMBIA0390∷GUS with report gene codingβ-D-glucuronidase（GUS）was transferred into callus by A.tumefaciens mediated transformation.The results of GUS activity proved that the improved transient expression system in callus of grapes contributed to the expression of exogenous genes in the callus of grapes,which provides technical support for the study of gene function of grapevine.